研究者総覧

仁木 利郎 (ニキ トシロウ)

  • 病理学講座(統合病理学部門) 教授
Last Updated :2021/09/22

研究者情報

学位

  • 医学博士(東京大学)

ホームページURL

J-Global ID

研究キーワード

  • 微小環境   筋線維芽細胞   上皮間質相互作用   分子標的   肺がん   Microenvironment   Fibroblast   Epithelial-stromal interaction   Molecular target therapy   Lung cancer   

研究分野

  • ライフサイエンス / 人体病理学

経歴

  • 2006年 - 現在  自治医科大学病理学講座 教授
  • 2005年 - 現在  Professor, Jichi Medical University
  • 2001年 - 2004年  東京大学人体病理学・病理診断学分野 助教授
  • 2001年 - 2004年  Associate Prof., University of Tokyo
  • 1997年 - 2000年  国立がんセンター主任研究官
  • 1997年 - 2000年  Research Staff, National Cancer Center
  • 1991年 - 1996年  ベルギー自由大学研究員
  • 1991年 - 1996年  Research Fellow, Free University of Brussel
  • 1987年 - 1990年  東京大学助手
  • 1987年 - 1990年  Research Associate, University of Tokyo
  • 自治医科大学(JMU)教授

学歴

  • 1983年04月 - 1986年03月   東京大学   医学系研究科   人体病理
  •         - 1986年   東京大学   Graduate School, Division of Medical Sciences   Human Pathology
  • 1979年04月 - 1983年03月   東京大学   医学部   医学科
  •         - 1983年   東京大学   Faculty of Medicine   school of Medicine

所属学協会

  • International Association for the Study of Lung Cancer   American Association for Cancer Research   日本癌学会   日本肺癌学会   日本臨床細胞学会   日本病理学会   International Association for the Study of Lung Cancer   The American Association for Cancer Research   The Japan Society of Pathology   The Japan Cancer Association   The Japan Lung Cancer Society   The Japanese Society of Clinical Cytology   The Japanese Society of Pathology   

研究活動情報

論文

  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 蘆澤 健太郎, 河田 浩敏, 金井 信行, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 439  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 天野 雄介, 松原 大祐, 吉本 多一郎, 仁木 利郎
    日本病理学会会誌 105 1 419  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 吉本 多一郎, 松原 大祐, 福嶋 敬宜, 仁木 利郎
    日本病理学会会誌 105 1 391  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • 森田 剛平, 三登 久美子, 仁木 利郎, 福嶋 敬宜
    日本病理学会会誌 105 1 367  (一社)日本病理学会 2016年04月 [査読無し][通常論文]
  • Fukusato T, Soejima Y, Kondo F, Inoue M, Watanabe M, Takahashi Y, Aso T, Uozaki H, Sano K, Sanada Y, Niki T
    Hepatology research : the official journal of the Japan Society of Hepatology 45 10 E32 - E42 2015年10月 [査読有り][通常論文]
     
    AimRecent studies have indicated that hepatocellular adenoma (HCA) is a heterogenous group of benign tumors with various genetic and clinicopathological characteristics. We delineated the clinicopathological characteristics of HCA subtypes and evaluated the expression of transporter protein OATP1B3 in HCA. MethodsHCA in 34 Japanese patients were investigated immunohistochemically and classified into four subtypes (HNF1-inactivated type, H-HCA; -catenin-activated type, b-HCA; inflammatory type, I-HCA; unclassified type, u-HCA). Immunostaining of OATP1B3 protein in HCA tissue sections was performed to determine the association between OATP1B3 expression and HCA subtypes. ResultsHCA was categorized into the following four subtypes and two combined subtypes: 10 H-HCA (29%), 10 I-HCA (29%), seven b-HCA (21%), two b-HCA/H-HCA (6%), two b-HCA/I-HCA (6%) and three u-HCA (9%). The male-to-female ratio was 18:16. Oral contraceptive use was rare but seven HCA were found in patients with glycogen storage disease, congenital absence of the portal vein and idiopathic portal hypertension. OATP1B3 expression was decreased in 24 HCA but was preserved or increased in 10 HCA. All nine HCA with nuclear staining for -catenin showed preserved or enhanced OATP1B3 expression, indicating a significant association between nuclear -catenin accumulation and OATP1B3 expression in HCA. ConclusionHCA subtype classification was validated in 91% of our Japanese subjects although their clinical backgrounds including rare contraceptive use were different from European subjects. A close association between preserved or enhanced OATP1B3 expression and b-HCA subtype indicated important modalities for clinical decisions in the treatment and follow up of patients with HCA.
  • Yukihiro Sanada, Koichi Mizuta, Toshiro Niki, Masahisa Tashiro, Yuta Hirata, Noriki Okada, Naoya Yamada, Yoshiyuki Ihara, Taizen Urahashi, Yurie Soejima, Toshio Fukusato, Fukuo Kondo
    JOURNAL OF HEPATO-BILIARY-PANCREATIC SCIENCES 22 10 746 - 756 2015年10月 [査読有り][通常論文]
     
    Background Hepatocellular nodules caused by congenital extrahepatic portosystemic shunts (CEPS) occur as a result of abnormal portal blood flow, and are mostly cases of benign focal nodular hyperplasia (FNH). However, hepatocellular adenomas (HCA) and hepatocellular carcinomas have been documented in the CEPS patients. HCA can now be immunohistochemically diagnosed; therefore, the concept of hepatocellular nodules resulting from CEPS should be revisited. In this study, we performed a retrospective immunohistochemical investigation of hepatocellular nodules from livers isolated from the CEPS patients undergoing living donor liver transplantation (LDLT). Methods Hepatocellular nodules from livers of five patients with CEPS who underwent LDLT between June 2004 and October 2012 at our institution were immunohistochemically investigated. HCA were classified into four subtypes (HNF1 alpha-inactivated HCA (H-HCA); inflammatory HCA; beta-catenin-activated HCA (b-HCA); unclassified HCA). Results Sixteen hepatocellular nodules were collected from livers of five patients with CEPS who underwent LDLT. Ten hepatocellular nodules were categorized as FNH (62.5%), five were categorized as b-HCA (31.3%), and one was categorized as H-HCA (6.2%). Conclusions Some of the hepatocellular nodules resulting from CEPS were indicative of HCAs, especially the b-HCA subtype which has the potential for malignant transformation. Surgical or interventional treatments might have to be performed when hepatocellular nodules appear in the CEPS patients.
  • Shin Saito, Kazue Morishima, Takashi Ui, Hiroko Hoshino, Daisuke Matsubara, Shumpei Ishikawa, Hiroyuki Aburatani, Masashi Fukayama, Yoshinori Hosoya, Naohiro Sata, Alan K. Lefor, Yoshikazu Yasuda, Toshiro Niki
    BMC CANCER 15 82  2015年02月 [査読有り][通常論文]
     
    Background: Although advanced esophageal squamous-cell carcinoma (ESCC) is treated using a multidisciplinary approach, outcomes remain unsatisfactory. The microenvironment of cancer cells has recently been shown to strongly influence the biologic properties of malignancies. We explored the effect of supernatant from esophageal fibroblasts on the cell growth and chemo-resistance of ESCC cell lines. Methods: We used 22 ESCC cell lines, isolated primary human esophageal fibroblasts and immortalized fibroblasts. We first examined cell proliferation induced by fibroblast supernatant. The effect of supernatant was evaluated to determine whether paracrine signaling induced by fibroblasts can influence the proliferation of cancer cells. Next, we examined the effects of adding growth factors HGF, FGF1, FGF7, and FGF10, to the culture medium of cancer cells. These growth factors are assumed to be present in the culture supernatants of fibroblasts and may exert a paracrine effect on the proliferation of cancer cells. We also examined the intrinsic role of HGF/MET and FGFs/FGFR in ESCC proliferation. In addition, we examined the inhibitory effect of lapatinib on ESCC cell lines and studied whether the fibroblast supernatants affect the inhibitory effect of lapatinib on ESCC cell proliferation. Finally, we tested whether the FGFR inhibitor PD-173074 could eliminate the rescue effect against lapatinib that was induced by fibroblast supernatants. Results: The addition of fibroblast supernatant induces cell proliferation in the majority of cell lines tested. The results of experiments to evaluate the effects of adding growth factors and kinase inhibitors suggests that the stimulating effect of fibroblasts was attributable in part to HGF/MET or FGF/FGFR. The results also indicate diversity in the degree of dependence on HGF/MET and FGF/FGFR among the cell lines. Though lapanitib at 1 mu M inhibits cell proliferation by more than 50% in the majority of the ESCC cell lines, fibroblast supernatant can rescue the growth inhibition of ESCC cells. However, the rescue effect is abrogated by co-treatment with FGFR inhibitor. Conclusion: These results demonstrate that cell growth of ESCC depends on diverse receptor tyrosine kinase signaling, in both cell-autonomous and cell-non-autonomous manners. The combined inhibition of these signals may hold promise for the treatment of ESCC.
  • Daisuke Matsubara, Yuka Kishaba, Taichiro Yoshimoto, Yuji Sakuma, Takashi Sakatani, Tomoko Tamura, Shunsuke Endo, Yukihiko Sugiyama, Yoshinori Murakami, Toshiro Niki
    PATHOLOGY INTERNATIONAL 64 11 560 - 568 2014年11月 [査読有り][通常論文]
     
    We performed an immunohistochemical analysis of the expression of zinc-finger E-box binding homeobox 1 (ZEB1), a master regulator of epithelial-mesenchymal transition (EMT), and determined its relationship with E-cadherin in 157 non-small cell lung carcinomas (93 adenocarcinomas, 36 squamous cell carcinomas, 18 large cell carcinomas, and 10 pleomorphic carcinomas). Although the expression of E-cadherin was low in the subset of adenocarcinomas (10%) and squamous cell carcinomas (11%), ZEB1 expression was only observed in one case of squamous cell carcinoma and none of the adenocarcinomas. In contrast, the low expression of E-cadherin (50% and 90%, respectively) and the positive expression of ZEB1 (11% and 50%, respectively) were more frequently observed in poorly differentiated carcinomas (large cell carcinomas and pleomorphic carcinomas). Overall, the expression of ZEB1 was inversely correlated with that of E-cadherin. Furthermore, the distribution of ZEB1-positive cancer cells was more restricted than in the area in which the expression of E-cadherin was lost, and the former was detected within the latter. We concluded that the expression of ZEB1 was not necessarily associated with the low expression of E-cadherin in lung adenocarcinomas and squamous cell carcinomas. The expression of ZEB1 correlated with an undifferentiated and/or sarcomatoid morphology that may occur in the late stage of EMT.
  • Hideyuki Ohzawa, Takashi Sakatani, Toshiro Niki, Yoshikazu Yasuda, Yasuo Hozumi
    BREAST CANCER 21 5 563 - 570 2014年09月 [査読有り][通常論文]
     
    The effectiveness of neoadjuvant chemotherapy is evaluated on the basis of pathological responses and survival outcome, because achievement of a pathological complete response (pCR) is a good predictor of long-term survival. However, few studies have assessed the survival of breast cancer patients who received neoadjuvant chemotherapy including trastuzumab. The records of 161 breast cancer patients who received neoadjuvant chemotherapy between January 2006 and December 2011 were retrospectively reviewed. The patients were categorized into 4 subgroups on the basis of the status of the estrogen receptor (ER), the progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). HER2-positive patients received trastuzumab-based regimens. Pathological responses and survival were analyzed on the basis of breast cancer subtypes. The pCR results obtained were: luminal A and B (ER and/or PR-positive, HER2-negative), 6.3 % (5/79 cases); luminal-HER2 hybrid (ER and/or PR-positive, HER2-positive), 25.0 % (5/20 cases); HER2-enriched (ER and PR-negative, HER2-positive), 63.0 % (17/27 cases); and triple-negative (ER and PR-negative, HER2-negative), 25.7 % (9/35 cases). Achievement of pCR was a good predictor of disease-free survival in the HER2-enriched group. Overall survival of patients with pCR was slightly, but not significantly, better in the HER2-enriched and triple-negative subgroups. Responses and survival after neoadjuvant chemotherapy including trastuzumab of patients with HER2-positive tumors differed among disease subtypes. Our findings suggest that disease subtype is an important determinant of the efficacy of neoadjuvant chemotherapy.
  • Reem Ibrahim, Daisuke Matsubara, Wael Osman, Teppei Morikawa, Akiteru Goto, Shigeki Morita, Shumpei Ishikawa, Hiroyuki Aburatani, Daiya Takai, Jun Nakajima, Masashi Fukayama, Toshiro Niki, Yoshinori Murakami
    HUMAN PATHOLOGY 45 7 1397 - 1405 2014年07月 [査読有り][通常論文]
     
    Although protein arginine methyltransferase 5 (PRMT5) has been implicated in various cancers, its expression pattern in lung adenocarcinoma cell lines and tissues has not been elucidated enough. In this study, microarray analysis of 40 non-small-cell lung carcinoma cell lines showed that PRMT5 was a candidate histone methyltransferase gene that correlated with epithelial-mesenchymal transition. Immunocytochemical analysis of these cell lines indicated that the expression of PRMT5 was localized to the cytoplasm of E-cadherin-low and vimentin-high cell lines, whereas it was predominant in the nucleus and faint in the cytoplasm of E-cadherin-high and vimentin-low cell lines Immunohistochemical analysis of lung adenocarcinoma cases (n = 130) revealed that the expression of PRMT5 was high in the cytoplasm of 47 cases (36%) and the nuclei of 34 cases (26%). The marked cytoplasmic expression of PRMT5 was frequently observed in high-grade subtypes (1 of 17 low grade, 21 of 81 intermediate grade, and 25 of 32 high grade; P <.0001) such as solid adenocarcinoma with the low expression of thyroid transcription factor 1 (the master regulator of lung) and low expression of cytokeratin 7 and E-cadherin (2 markers for bronchial epithelial differentiation), whereas the high nuclear expression of PRMT5 was frequently noted in adenocarcinoma in situ, a low-grade subtype (6 of 17 low grade, 25 of 81 intermediate grade, and 3 of 32 high grade; P = .0444). The cytoplasmic expression of PRMT5 correlated with a poor prognosis (P =.0089). We herein highlighted the importance of PRMT5 expression, especially its cytoplasmic expression, in the process of epithelial-mesenchymal transition and loss of the bronchial epithelial phenotype of lung adenocarcinoma. (C) 2014 Elsevier Inc. All rights reserved.
  • Shin Saito, Kazue Morishima, Takashi Ui, Daisuke Matsubara, Tomoko Tamura, Sachiko Oguni, Yoshinori Hosoya, Naohiro Sata, Alan T. Lefor, Yoshikazu Yasuda, Toshiro Niki
    ONCOLOGY REPORTS 32 1 348 - 354 2014年07月 [査読有り][通常論文]
     
    The growth, invasiveness and metastasis of human cancers are determined not only by cancer cells, but also by their microenvironment. Activated stromal fibroblasts promote tumor progression by secreting growth factors. In the present study, we focused on interrelations between cancer and fibroblasts, the main component of tumor stroma. We retrospectively analyzed the relations of mortality to clinical, pathological, and a-smooth muscle actin (alpha-SMA) characteristics in 97 consecutive patients with esophageal squamous cell carcinoma (ESCC). In vitro, we used TE-11, KYSE150 and KYSE220 ESCC cell lines and isolated esophageal stromal fibroblasts, some of which were immortalized. Migration assays were conducted to assess the effects of fibroblasts on cancer-cell migration and 3-dimensional organotypic cultures. In vivo, TE-11 and KYSE220 cells plus immortalized fibroblasts were co-transplanted subcutaneously in Nod/Scid mice to assess the effects of fibroblasts on tumorigenicity. Clinicopathologically, the a-SMA expression of cancer stroma was correlated with venous invasion (p<0.01), nodal involvement (p=0.02), recurrence (p=0.01), and was a predictor of survival in patients with stage I and II ESCC (p=0.04). In vitro, the presence of fibroblasts strongly promoted the migration of TE-11, KYSE150 and KYSE220 cells. On organotypic culture, stromal invasion was observed only in the presence of immortalized fibroblasts. In vivo, tumors developed or grew in a fibroblast-dependent manner after implantation. Our findings provide evidence that stromal fibroblasts and tumor cells interact to promote tumor progression in ESCC. In patients with earlier stage ESCC, a-SMA may be a predictor of mortality. Inhibition of paracrine systems associated with tumor fibroblasts may slow or reverse tumor progression, potentially leading to the development of new targeted therapies.
  • Takashi Ui, Kazue Morishima, Shin Saito, Yuji Sakuma, Hirofumi Fujii, Yoshinori Hosoya, Shumpei Ishikawa, Hiroyuki Aburatani, Masashi Fukayama, Toshiro Niki, Yoshikazu Yasuda
    ONCOLOGY REPORTS 31 2 619 - 624 2014年02月 [査読有り][通常論文]
     
    Although cisplatin (CDDP) is a key drug in the treatment of esophageal squamous cell carcinoma (ESCC), acquired chemoresistance remains a major problem. Combination therapy may represent one strategy to overcome this resistance. Heat shock protein 90 (HSP90) is known to be overexpressed in several types of cancer cells, and its inhibition by small molecules, either alone or in combination, has shown promise in the treatment of solid malignancies. In the present study, we evaluated the synergistic effects of combining CDDP with the HSP90 inhibitor 17-N-allylamino-17-demethoxy geldanamycin (17-AAG) on two CDDP-resistant human esophageal squamous cancer cell lines, KYSE30 and KYSE150. The results obtained demonstrated the synergistic inhibitory effects of CDDP and 17-AAG on the growth of KYSE30 and KYSE150 cells. Cell growth and cell number were more effectively reduced by the combined treatment with CDDP and 17-AAG than by the treatment with either CDDP or 17-AAG alone. Western blotting revealed that the combined action of CDDP and 17-AAG cleaved poly (ADP-ribose) polymerase (PARP) and caspase-3, which demonstrated that the reduction in both cell growth and cell number was mediated by apoptosis. Time-course experiments showed that reduction in X-linked inhibitor of apoptosis protein (XIAP) and phosphorylated Akt were concomitant with apoptosis. The results of the present study demonstrate that 17-AAG synergizes with CDDP and induces apoptosis in CDDP-resistant ESCC cell lines, and also that modulation of the Akt/XIAP pathway may underlie this synergistic effect. Combination therapy with CDDP and an HSP90 inhibitor may represent a promising strategy to overcome CDDP resistance in ESCC.
  • Yuji Sakuma, Shoichi Matsukuma, Yoshiyasu Nakamura, Mitsuyo Yoshihara, Shiro Koizume, Hironobu Sekiguchi, Haruhiro Saito, Haruhiko Nakayama, Yoichi Kameda, Tomoyuki Yokose, Sachiko Oguni, Toshiro Niki, Yohei Miyagi
    Laboratory Investigation 93 10 1137 - 1146 2013年10月 [査読有り][通常論文]
     
    Lung cancers harboring epidermal growth factor receptor (EGFR) mutations depend on constitutive activation of the kinase for survival. Although most EGFR-mutant lung cancers are sensitive to EGFR tyrosine kinase inhibitors (TKIs) and shrink in response to treatment, acquired resistance to TKI therapy is common. We demonstrate here that two EGFR-mutated lung adenocarcinoma cell lines, HCC827 and HCC4006, contain a subpopulation of cells that have undergone epithelial-to-mesenchymal transition and survive independent of activated EGFR. These EGFR-independent cancer cells, herein termed gefitinib-resistant (GR) cells, demonstrate higher levels of basal autophagy than their parental cells and thrive under hypoxic, reduced-serum conditions in vitro this somewhat simulates the hypoxic environment common to cancerous tissues. We show that depletion of the essential autophagy gene, ATG5, by small interfering RNA (siRNA) or chloroquine, an autophagy inhibitor, markedly reduces GR cell viability under hypoxic conditions. Moreover, we show a significant elevation in caspase activity in GR cells following knockdown of ATG5. These results suggest that GR cells can evade apoptosis and survive in hostile, hypoxic environments with constant autophagic flux. We also show the presence of autophagosomes in some cancer cells from patient samples, even in untreated EGFR-mutant lung cancer tissue samples. Together, our results indicate that autophagy inhibitors alone or in combination with EGFR TKIs may be an effective approach for the treatment of EGFR-mutant lung cancers, where basal autophagy of some cancer cells is upregulated. © 2013 USCAP.
  • Daisuke Matsubara, Yuka Kishaba, Shumpei Ishikawa, Takashi Sakatani, Sachiko Oguni, Tomoko Tamura, Hiroko Hoshino, Yukihiko Sugiyama, Shunsuke Endo, Yoshinori Murakami, Hiroyuki Aburatani, Masashi Fukayama, Toshiro Niki
    CANCER SCIENCE 104 2 266 - 273 2013年02月 [査読有り][通常論文]
     
    BRG1 and BRM, two core catalytic subunits in SWI/SNF chromatin remodeling complexes, have been suggested as tumor suppressors, yet their roles in carcinogenesis are unclear. Here, we present evidence that loss of BRG1 and BRM is involved in the progression of lung adenocarcinomas. Analysis of 15 lung cancer cell lines indicated that BRG1 mutations correlated with loss of BRG1 expression and that loss of BRG1 and BRM expression was frequent in E-cadherin-low and vimentin-high cell lines. Immunohistochemical analysis of 93 primary lung adenocarcinomas showed loss of BRG1 and BRM in 11 (12%) and 16 (17%) cases, respectively. Loss of expression of BRG1 and BRM was frequent in solid predominant adenocarcinomas and tumors with low thyroid transcription factor-1 (TTF-1, master regulator of lung) and low cytokeratin7 and E-cadherin (two markers for bronchial epithelial differentiation). Loss of BRG1 was correlated with the absence of lepidic growth patterns and was mutually exclusive of epidermal growth factor receptor (EGFR) mutations. In contrast, loss of BRM was found concomitant with lepidic growth patterns and EGFR mutations. Finally, we analyzed the publicly available dataset of 442 cases and found that loss of BRG1 and BRM was frequent in E-cadherin-low, TTF-1-low, and vimentin-high cases and correlated with poor prognosis. We conclude that loss of either or both BRG1 and BRM is involved in the progression of lung adenocarcinoma into solid predominant tumors with features of epithelial mesenchymal transition and loss of the bronchial epithelial phenotype. BRG1 loss was specifically involved in the progression of EGFR wild-type, but not EGFR-mutant tumors.
  • Daisuke Matsubara, Yoshihiko Kanai, Shumpei Ishikawa, Shiori Ohara, Taichiro Yoshimoto, Takashi Sakatani, Sachiko Oguni, Tomoko Tamura, Hiroaki Kataoka, Shunsuke Endo, Yoshinori Murakami, Hiroyuki Aburatani, Masashi Fukayama, Toshiro Niki
    JOURNAL OF THORACIC ONCOLOGY 7 12 1872 - 1876 2012年12月 [査読有り][通常論文]
     
    Rearranged during transfection (RET) fusions have been newly identified in approximately 1% of patients with primary lung tumors. However, patient-derived lung cancer cell lines harboring RET fusions have not yet been established or identified, and therefore, the effectiveness of an RET inhibitor on lung tumors with endogenous RET fusion has not yet been studied. In this study, we report identification of CCDC6-RET fusion in the human lung adenocarcinoma cell line LC-2/ad. LC-2/ad showed distinctive sensitivity to the RET inhibitor, vandetanib, among 39 non-small lung cancer cell lines. The xenograft tumor of LC-2/ad showed cribriform acinar structures, a morphologic feature of primary RET fusion-positive lung adenocarcinomas. LC-2/ad cells could provide useful resources to analyze molecular functions of RET-fusion protein and its response to RET inhibitors.
  • Shin Saito, Yoshinori Hosoya, Kazue Morishima, Takashi Ui, Hidenori Haruta, Kentaro Kurashina, Yoshiyuki Meguro, Toru Zuiki, Naohiro Sata, Hirofumi Fujii, Daisuke Matsubara, Toshiro Niki, Alan T. Lefor, Yoshikazu Yasuda
    JOURNAL OF DIGESTIVE DISEASES 13 8 407 - 413 2012年08月 [査読有り][通常論文]
     
    OBJECTIVE: Adenosquamous carcinoma originating in the stomach is an unusual neoplasm with few existing histological studies. This study was aimed to gain insight into the histogenetic and clinicopathological characteristics of gastric cancer with squamous cell carcinoma (SCC) components. METHODS: From January 2001 to June 2010 a total of 1735 patients underwent a resection of gastric cancer. Histopathologically, eight patients had adenocarcinoma containing SCC components, in which the proportion of SCC components was above 25% of the total tumor mass in four patients. The immunohistochemical and clinicopathological characteristics of these eight patients were analyzed. RESULTS: The median survival duration was 22 months. Adenocarcinoma was present at the superficial layer of all tumors and SCC was primarily present at sites with deep invasion. Immunohistochemically, adenocarcinoma components were positive for cytokeratin (CK) 8/18/19 and CK7 in all cases. SCC components were positive for carcinoembryonic antigen and CK7 in more than 60% of patients. Expression patterns of p53 product were identical in both components. SCC components were positive for 34 beta E12 and adenocarcinoma components were negative for 34 beta E12 in all patients. CONCLUSIONS: SCC components are derived from squamous metaplasia in a pre-existing adenocarcinoma. A gastric adenocarcinoma with SCC components is associated with various patterns of metastasis and both SCC and adenocarcinoma components have the potential for metastasis. Gastric cancer with SCC components is a clinically aggressive tumor.
  • Daisuke Matsubara, Toshiro Niki
    JOURNAL OF THORACIC ONCOLOGY 7 4 771 - 772 2012年04月 [査読有り][通常論文]
  • Akiteru Goto, Chih-Ping Li, Satoshi Ota, Toshiro Niki, Yuji Ohtsuki, Shinichi Kitajima, Suguru Yonezawa, Chihaya Koriyama, Suminori Akiba, Hisataka Uchima, Yueh-Min Lin, Kun-Tu Yeh, Jae-Soo Koh, Chul-Woo Kim, Kun-Yong Kwon, Min En Nga, Masashi Fukayama
    JOURNAL OF MEDICAL VIROLOGY 83 8 1383 - 1390 2011年08月 [査読有り][通常論文]
     
    The role of human papillomavirus (HPV) in the development of lung and esophageal cancer remains inconclusive, which is in contrast to the established role HPV plays in the development of uterine cervical cancer. One of the reasons for this is the difference among reported HPV infection rates in these cancers. An analysis of 485 lung and esophageal cancers (176 lung squamous cell carcinoma, 128 lung adenocarcinoma, 181 esophageal carcinoma) in eight institutions in Asia (Tokyo, Kochi, Kagoshima, and Okinawa, Japan; Seoul and Daegu, Korea; Changhua, Republic of China (Taiwan); Singapore, Singapore) was carried out in order to clarify infection rates with HPV. Samples were examined in one laboratory of the Department of Pathology, the University of Tokyo, Japan in order to avoid inter-laboratory variation using a combination of polymerase chain reaction and in situ hybridization (ISH). HPV was found in 6.3%, 7%, and 9.4% of patients with lung squamous cell carcinoma, lung adenocarcinoma, and esophageal cancer, respectively. Among the geographic areas surveyed, Kagoshima exhibited a significantly higher prevalence of HPV infection in cases of esophageal carcinoma (24.1%). There was no geographical difference in the infection rates of HPV in lung carcinomas. Subtype-specific ISH was also performed, which identified the high-risk HPV types 16/18 in the majority (75.7%) of the patients with lung and esophageal cancer positive for HPV. J. Med. Virol. 83:1383-1390, 2011. (C) 2011 Wiley-Liss, Inc.
  • Hiroshi Terasaki, Seiya Kato, Yoshihiro Matsuno, Masahiko Kusumoto, Toshiro Niki, Akihiro Hayashi, Kinuyo Terasaki, Naofumi Hayabuchi
    JOURNAL OF THORACIC IMAGING 26 1 74 - 81 2011年02月 [査読有り][通常論文]
     
    "Adenocarcinoma, mixed subtype'' is the most common histologic subtype of lung adenocarcinomas in the World Health Organization classification of 2004. For small peripheral adenocarcinomas, for example, those measuring 2 cm or less in greatest diameter, invasive areas can present various histologic patterns. The purpose of this study is to present the radiologic features of small peripheral lung adenocarcinomas with or without a bronchioloalveolar component and with or without invasive areas, in comparison with histopathologic findings. For this purpose, a detailed evaluation of the characteristics of solid regions in ground-glass opacity on high-resolution computerized tomographic images of lung adenocarcinoma is useful.
  • Daisuke Matsubara, Shumpei Ishikawa, Oguni Sachiko, Hiroyuki Aburatani, Masashi Fukayama, Toshiro Niki
    AMERICAN JOURNAL OF PATHOLOGY 177 5 2191 - 2204 2010年11月 [査読有り][通常論文]
     
    Epidermal growth factor receptor (EGFR) and MET are molecular targets for lung cancer treatment. The relationships between expression, activation, and gene abnormalities of these two targets are currently unclear. Here, we demonstrate that a panel of 40 lung cancer cell lines could be classified into two groups. Group I was characterized by (1) high phosphorylations of MET and EGFR, (2) frequent mutation or amplification of EGFR, MET, and human epidermal growth factor receptor-2 (HER2), (3) high expressions of bronchial epithelial markers (thyroid transcription factor-1 (TTF-1), MUG!, and Cytokeratin 7 (CK7)); and (4) high expressions of MET, human epidermal growth factor receptor-3, E-cadherin, cyclooxygenase-2, and laminin gamma2. In contrast, Group 11 exhibited little or no phosphorylation of MET and EGFR; no mutation or amplification of EGFR, MET, and HER2; were triple-negative for TTF-1, MUC1, and CK7; and showed high expressions of vimentin, fibroblast growth factor receptor-1, and transcription factor 8. Importantly, Group I was more sensitive to gefitinib and more resistant to cisplatin and paclitaxel than Group II. The clinical relevance was confirmed in publicly available data on 442 primary lung adenocarcinoma patients; survival benefits by postoperative chemotherapy were seen in only patients with tumors corresponding to Group II. Overall, co-activation of EGFR and MET defines a distinct subgroup of lung carcinoma with characteristic genetic abnormalities, gene expression pattern, and response to chemotherapeutic reagents. (Am J Pathol 2010, 177:2191-2204 DOI: 10.2353/ajpath.2010.100217)
  • Matsubara D, Ishikawa S, Oguni S, Aburatani H, Fukayama M, Niki T
    Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer 5 1317 - 1324 9 2010年09月 [査読有り][通常論文]
  • Kazuha Sakamoto, Shinji Sakurai, Tatsuo Kanda, Yuji Sakuma, Tsunekazu Hishima, Mitsugu Hironaka, Takeo Bamba, Yoshiko Keira, Yasuo Takano, Toshiro Niki, Tadashi Hasegawa, Seiichi Hirota
    CANCER SCIENCE 101 5 1270 - 1278 2010年05月 [査読有り][通常論文]
     
    (Cancer Sci 2010; 101: 1270-1278).
  • Yuka Kishaba, Daisuke Matsubara, Toshiro Niki
    PATHOLOGY INTERNATIONAL 60 5 378 - 385 2010年05月 [査読有り][通常論文]
     
    In this study we explored the distribution of nestin-positive cells in extraneural human tissues with special reference to stromal myofibroblasts. Tissue microarrays were constructed from various tissues with normal histology and tissues with fibrosing disorders. Sections were immunostained for nestin, alpha-smooth muscle actin (alpha-SMA), desmin, vimentin, CD34, and other stromal markers. Nestin was expressed in the myoepithelium of the breast, podocytes of the renal glomerulus, and endothelial cells of most organs. Nestin was also expressed in the stroma of several organs, including the intestine, uterine cervix, and endometrium. Nestin-positive fibroblast-like cells appeared in the stroma of the kidney, pancreas, lung, and skin in fibrosing conditions. With the notable exception of endometrial stromal cells, most of these nestin-positive stromal cells were alpha-SMA-positive. Interestingly, we observed a concomitant appearance of nestin- and CD34-positive myofibroblasts under fibrosing conditions. Further investigation showed that nestin was expressed by stromal fibroblasts in cervical squamous cell carcinoma, but not in lung adenocarcinoma, pointing to heterogeneity of cancer stroma. In conclusion, nestin was expressed in variable proportions of stromal myofibroblasts in human tissues. The differential expression of nestin may indicate phenotypic and functional heterogeneity. Nestin-positive myofibroblast may represent a relatively immature subpopulation of cells with multipotentiality.
  • Daisuke Matsubara, Teppei Morikawa, Akiteru Goto, Jun Nakajima, Masashi Fukayama, Toshiro Niki
    MODERN PATHOLOGY 22 6 776 - 785 2009年06月 [査読有り][通常論文]
     
    We report here the presence of subepithelial myofibroblasts in pure bronchioloalveolar carcinoma and a subset of invasive lung adenocarcinoma. The subepithelial myofibroblasts we describe were observed in a peculiar location beneath the cancer cells in the alveolar septa. Immunohistochemically, they were positive for alpha-smooth muscle actin and calponin, but negative for desmin and h-caldesmon. To gain insight into their biological significance, we examined 116 surgically resected lung adenocarcinomas. The resected tumors included 13 bronchioloalveolar carcinomas, 20 mixed type adenocarcinomas with bronchioloalveolar carcinoma components, 57 papillary adenocarcinomas, 22 solid adenocarcinomas with mucin, and 4 acinar adenocarcinomas. All specimens were immunostained for alpha-smooth muscle actin to visualize the myofibroblasts. In all of the pure bronchioloalveolar carcinomas observed, the subepithelial myofibroblasts were completely preserved adjacent to the cancer cells. In mixed adenocarcinomas with bronchioloalveolar carcinoma components, subepithelial myofibroblasts were present in the bronchioloalveolar carcinoma components, but scanty in the invasive areas, where stromal myofibroblasts emerged between the cancer cell nests. Subepithelial myofibroblasts were retained, however, in the invasive areas of a subset of invasive adenocarcinomas. Survival analysis showed that the retention of subepithelial myofibroblasts in these invasive tumors was associated with low rates of lymphatic and vascular invasion, a low rate of lymph node involvement, and an excellent patient survival. These results suggest that subepithelial myofibroblasts increase in bronchioloalveolar carcinomas, but are gradually replaced by typical stromal myofibroblasts during progression into invasive cancer. A subset of invasive adenocarcinomas retains subepithelial myofibroblasts. Analysis of subepithelial myofibroblasts may be helpful in identifying a subset of lung adenocarcinoma with excellent prognosis. Modern Pathology (2009) 22, 776-785; doi: 10.1038/modpathol.2009.27; published online 27 March 2009
  • Hidenori Haruta, Yoshinori Hosoya, Kazuya Sakuma, Hiroyuki Shibusawa, Kiichi Satoh, Hironori Yamamoto, Akira Tanaka, Toshiro Niki, Kentaro Sugano, Yoshikazu Yasuda
    JOURNAL OF DIGESTIVE DISEASES 9 4 213 - 218 2008年11月 [査読有り][通常論文]
     
    BACKGROUND: The endoscopic resection of early gastric cancers (EGC) is a standard technique in Japan and is increasingly used throughout the world. Further experience in the treatment of EGC and a clearer delineation of the factors related to lymph-node metastasis would permit a more accurate assessment of endoscopic resection. METHODS: The study group comprised 1389 patients with EGC who underwent gastrectomy with lymph-node dissection. We evaluated the relations of lymph-node metastasis to clinicopathological factors. RESULTS: Of the 718 patients with intramucosal carcinomas, 14 (1.9%) had lymph-node metastasis. All cases of lymph-node metastasis were associated with ulceration. No lymph-node metastasis was found in patients with intramucosal carcinomas without ulceration, irrespective of tumor size and histological type. Lymph-node metastasis was present in 14 (4.7%) of the 296 patients who had cancer with a submucosal invasion depth of less than 500 mu m (sm 1). Significantly increased rates of lymph-node metastasis were associated with undifferentiated types, ulcerated lesions and lymphatic invasion. No lymph-node metastasis was found in patients with differentiated sm 1 carcinomas 30 mm or less in diameter without ulceration. Lymph-node metastasis occurred in 29% of the patients who had cancer with a submucosal invasion depth of 500 mu m or more (sm 2). CONCLUSION: This large series of patients with EGC provides further evidence supporting the expansion of indications for endoscopic treatment, as well as warns against potential risks.
  • Yu Nakamura, Daisuke Matsubara, Akiteru Goto, Satoshi Ota, Oguni Sachiko, Shumpei Ishikawa, Hiroyuki Aburatani, Keiji Miyazawa, Masashi Fukayama, Toshiro Niki
    CANCER SCIENCE 99 1 14 - 22 2008年01月 [査読有り][通常論文]
     
    In this study we explored the mechanisms of constitutive activation of c-Met in lung adenocarcinoma cell lines. First, we examined levels of c-Met and phospho-c-Met (Y1234/Y1235) in a panel of lung adenocarcinoma cell lines by Western blot analysis. c-Met expression was found in 12 of 14 cell lines and an overall correlation between the expressions of c-Met and phospho-c-Met was noted. c-Met was constitutively activated particularly at high levels in five cell lines (PC3, LC-2/ad, L27, H1648, and H2009). c-Met amplification was identified in L27 and H1648 by single nucleotide polymorphism array analysis, but no mutations were identified in the Sema domain or in any part of the cytoplasmic domain of c-Met. Experiments with neutralizing anti-hepatocyte growth factor (HGF) antibody, scatter assay using Madin-Darby canine kidney cells, and Western blotting on conditioned media of the cell lines revealed that the constitutive phosphorylation of c-Met was largely ligand-independent. The inhibition of cell-matrix adhesion induced the dephosphorylation of c-Met in the five cell lines tested. This was accompanied by downregulation of c-Met in three of the five cell lines. In contrast, the inhibition of cell-cell adhesion by neutralizing E-cadherin antibody had a minimal effect on the expression and phosphorylation of c-Met. These results reveal three features of the constitutive activation of c-Met in our panel of lung adenocarcinoma cell lines: (i) it correlates with c-Met overexpression, either with or without gene amplification; (ii) it is largely ligand-independent; and (iii) it depends on cell-matrix adhesion.
  • Inamura K, Takeuchi K, Togashi Y, Nomura K, Ninomiya H, Okui M, Satoh Y, Okumura S, Nakagawa K, Soda M, Choi YL, Niki T, Mano H, Ishikawa Y
    Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer 3 13 - 17 1 2008年01月 [査読有り][通常論文]
  • Chih-Ping Li, Akiteru Goto, Akira Watanabe, Kengo Murata, Satoshi Ota, Toshiro Niki, Hiroyuki Aburatani, Masashi Fukayama
    PATHOLOGY RESEARCH AND PRACTICE 204 5 295 - 304 2008年 [査読有り][通常論文]
     
    The incidence of lung cancer (LC) is markedly increased among patients with usual interstitial pneumonia (UIP), and tobacco smoking is its superimposed risk factor. AKRIB10 (aldo-keto reductase IB10) is frequently overexpressed in pulmonary squamous cell carcinoma and adenocarcinoma in smokers. To investigate the role of AKRIB10 in the pulmonary carcinogenesis in UIP with correlation to tobacco smoking, we examined 13 UIP cases with LC, 13 UIP cases without LC, and 30 cases of non-UIP LC using AKRIB10 immunohistochemistry. AKRIB10 immunoreactivity was confined to squamous metaplasia in honeycomb lesions of UIP and neoplastic cells of LC. Squamous metaplastic foci showed AKR1B10 immunoreactivity more frequently in UIP with LC (24/36 foci, 67%) than in UIP without LC (16/44 foci, 37%) (P<0.01). AKRIB10 expression in UIP was also more frequent in squamous metaplastic foci in smokers (38/67 foci, 57%) than in non-smokers (2/13 foci, 15%) (P<0.01). AKRIB10 expression was frequently observed in both UIP-associated LC (10/13 foci, 77%) and non-UIP LC (18/30 foci, 60%). Ki-67 labeling index was significantly higher in AKRIB10-positive squamous metaplasia of UIP than in AKRIB10-negative squamous metaplasia of UIP. Our results demonstrate that AKRIB10 is involved in the development of LC in UIP in association with smoking. AKR1B10 might be useful as a new marker for identification of high LC risk patients in UIP. (C) 2008 Elsevier GmbH. All rights reserved.
  • Kengo Murata, Satoshi Ota, Toshiro Niki, Akiteru Goto, Chih-Ping Li, Urbiztondo Maria Rhea Ruriko, Shumpei Ishikawa, Hiroyuki Aburatani, Takayuki Kuriyama, Masashi Fukayama
    EXPERIMENTAL AND MOLECULAR PATHOLOGY 83 3 367 - 376 2007年12月 [査読無し][通常論文]
     
    Idiopathic pulmonary fibrosis (IPF) is the most common lung disease predisposing lung cancer. To clarify the early phase of epithelial abnormalities in IPF, we used an in vitro squamous metaplasia model, transforming growth factor beta 1 (TGF beta 1)-treated airway epithelial cells (BEAS-2B). The model repeated the expression of squamous epithelial character, such as involucrin, and keratin 6 and 14. DNA microarray analysis disclosed a unique expression signature in TGF beta 1-treated airway epithelial cells, 20 specifically up-regulated genes including p63,jagged 1 (jag1) and the genes of structure proteins. Western blotting and RT-PCR analysis revealed that Delta Np63 alpha was the dominant isoform of p63 in our experimental model. Immunohistochemical analysis demonstrated the expression of p63 and jag1 in lung tissues of IPF. Inhibition of p63 with siRNA caused the down-regulation of jag1 expression, but not of involucrin, or keratin 6 and 14. Interestingly, the up-regulation of p63 was totally suppressed by N-acetyl-L-cysteine (NAC), but not by dexamethasone or pirfenidone. Thus, the p63-jag1 pathway may be up-regulated at an early phase of epithelial abnormalities in IPF, which can be overcome by NAC even in the TGF beta 1-rich milieu. (c) 2007 Elsevier Inc. All rights reserved.
  • Kengo Murata, Satoshi Ota, Toshiro Niki, Akiteru Goto, Chih-Ping Li, Urbiztondo Maria Rhea Ruriko, Shumpei Ishikawa, Hiroyuki Aburatani, Takayuki Kuriyama, Masashi Fukayama
    EXPERIMENTAL AND MOLECULAR PATHOLOGY 83 3 367 - 376 2007年12月 [査読有り][通常論文]
     
    Idiopathic pulmonary fibrosis (IPF) is the most common lung disease predisposing lung cancer. To clarify the early phase of epithelial abnormalities in IPF, we used an in vitro squamous metaplasia model, transforming growth factor beta 1 (TGF beta 1)-treated airway epithelial cells (BEAS-2B). The model repeated the expression of squamous epithelial character, such as involucrin, and keratin 6 and 14. DNA microarray analysis disclosed a unique expression signature in TGF beta 1-treated airway epithelial cells, 20 specifically up-regulated genes including p63,jagged 1 (jag1) and the genes of structure proteins. Western blotting and RT-PCR analysis revealed that Delta Np63 alpha was the dominant isoform of p63 in our experimental model. Immunohistochemical analysis demonstrated the expression of p63 and jag1 in lung tissues of IPF. Inhibition of p63 with siRNA caused the down-regulation of jag1 expression, but not of involucrin, or keratin 6 and 14. Interestingly, the up-regulation of p63 was totally suppressed by N-acetyl-L-cysteine (NAC), but not by dexamethasone or pirfenidone. Thus, the p63-jag1 pathway may be up-regulated at an early phase of epithelial abnormalities in IPF, which can be overcome by NAC even in the TGF beta 1-rich milieu. (c) 2007 Elsevier Inc. All rights reserved.
  • Yu Nakamura, Toshiro Niki, Akiteru Goto, Teppei Morikawa, Keiji Miyazawa, Jun Nakajima, Masashi Fukayama
    CANCER SCIENCE 98 7 1006 - 1013 2007年07月 [査読有り][通常論文]
     
    c-Met is often overexpressed in non-small cell lung cancer, but it remains unsolved whether its overexpression leads to its activation. We used an antibody specific to phospho-c-Met (Tyr1235) to investigate c-Met activation immunohistochemically in 130 surgically resected lung adenocarcinomas. The expression of c-Met and hepatocyte growth factor (HGF) was also investigated. Phospho-c-Met was positive in 21.5% (28/130) of cases. c-Met was positive in 74.6% of cases (97/130) and was expressed at high levels in 36.1% of cases (47/130). HGF was expressed at high levels in 31.5% of cases (41/130). Phospho-c-Met was correlated with high levels of HGF (P = 0.0010) and high levels c-Met expression (P = 0.0303), but it was also found to be positive in 12 cases with little to no HGF expression. Phospho-c-Met expression was significantly associated with tumor differentiation (P = 0.0023) and papillary histology (P = 0.0011), but not with pathological stage, lymph node metastasis or survival. High levels of c-Met and HGF were also associated with papillary histology (P = 0.0056 and P = 0.0396, respectively), but not with tumor differentiation. Phospho-c-Met was correlated with phospho-Akt (P = 0.0381), but not with phospho-Erk or phospho-Stat3. Phospho-Akt expression was marginally correlated with the expression of phospho-epidermal growth factor receptor (EGFR) (P = 0.0533) and, importantly, it was strongly correlated with the expression of either phospho-c-Met or phospho-EGFR (P = 0.0013). The data suggest that in lung adenocarcinoma tissue, c-Met activation may take place either ligand-dependently or ligand-independently via c-Met overexpression. c-Met activation may play special roles in the papillary subtype and in well differentiated lung adenocarcinomas.
  • Tao Wang, Toshiro Niki, Akiteru Goto, Satoshi Ota, Teppei Morikawa, Yu Nakamura, Etsuko Ohara, Shumpei Ishikawa, Hiroyuki Aburatani, Jun Nakajima, Masashi Fukayama
    CANCER SCIENCE 98 4 506 - 511 2007年04月 [査読有り][通常論文]
     
    Tumor hypoxia is associated with a malignant phenotype of cancer cells and poor patient prognosis. To investigate the role of hypoxia in tumor progression, we studied the effects of hypoxia in the A549 lung adenocarcinoma cell line. First, we showed that hypoxic treatment decreased cell-cell adhesion and induced a scattering of cancer cells. Concomitant with these morphological changes, the motility of cancer cells was increased, as demonstrated by the Boyden chamber assay. Then, we used oligonucleotide array analyses to identify the genes causally related to the hypoxia-induced motile phenotype. The results showed that the expression of approximately 100 genes was induced more than 5-fold by hypoxia. These included (among others) epidermal growth factor receptor (EGFR), as well as other well-known hypoxia-induced genes, such as vascular endothelial growth factor. Immunohistochemical analyses of primary lung adenocarcinomas confirmed the induction of EGFR in tumor cells in the vicinity of necrotic areas, a histological indicator of tumor hypoxia. Remarkably, the EGFR inhibitor AG1478 (10 mu M) completely blocked the increased cell motility induced by hypoxia. Thus, the present study demonstrates the importance of the EGFR pathway in the increased motility of cancer cells that occurs in a hypoxic tumor environment.
  • N Yamauchi, A Watanabe, M Hishinuma, K Ohashi, Y Midorikawa, Y Morishita, T Niki, J Shibahara, M Mori, M Makuuchi, Y Hippo, T Kodama, H Iwanari, H Aburatani, M Fukayama
    MODERN PATHOLOGY 18 12 1591 - 1598 2005年12月 [査読有り][通常論文]
     
    Expression profiling of hepatocellular carcinoma has demonstrated that glypican 3 (GPC3), a heparan sulfate proteoglycan anchored to the membrane, is expressed at a markedly elevated level in hepatocellular carcinoma. In this paper, two monoclonal antibodies against GPC3, GPC3-C02 and A1836A, were confirmed to specifically recognize GPC3molecule in cells from hepatocellular carcinoma and hepatoblastoma cell lines by immunoblotting, and both were confirmed to recognize different epitopes of the GPC3 molecule by epitope mapping. Then, we evaluated the feasibility of GPC3-immunohistochemistry in the pathological diagnosis of benign and malignant hepatocellular lesions by applying these monoclonal antibodies to formalin-fixed and paraffin-embedded specimens. The immunoreactivity turned out to be identical in the two monoclonal antibodies and was thus confirmed to represent the actual expression of the GPC3 molecule. The expression was observed in the fetal liver, but not in normal adult liver, liver cirrhosis or hepatitis except for a tiny focus of a regenerative nodule of fulminant hepatitis. Diffusely positive staining of GPC3 was observed in malignant hepatocytes in hepatoblastomas and in hepatocellular carcinomas (47/56, 84%). GPC3 expression was independent of the differentiation and size of the hepatocellular carcinoma. On the other hand, there was only weak and focal staining in low-grade (2/8) and high-grade dysplastic nodules (6/8). GPC3 immunoreactivity was detected in only one of 23 metastatic lesions of colorectal carcinoma, and its expression was entirely absent in the liver cell adenoma (0/7), carcinoid tumor (0/1), and cholangiocellular carcinoma (0/16). When compared with immunohistochemistry of hepatocyte antigen and alpha-fetoprotein, GPC3-immunohistochemistry was siginificantly much more specific and sensitive for hepatocellular carcinomas. Thus, GPC3 was confirmed to be one of the oncofetal proteins now attracting attention for their promise both as markers of hepatocellular carcinoma in routine histological examination and as targets in monoclonal antibody-based hepatocellular carcinoma therapy.
  • D Matsubara, T Niki, S Ishikawa, A Goto, E Ohara, T Yokomizo, CW Heizmann, H Aburatani, S Moriyama, H Moriyama, Y Nishimura, N Funata, M Fukayama
    CANCER SCIENCE 96 12 844 - 857 2005年12月 [査読有り][通常論文]
     
    Previous studies suggest that some S100 proteins are involved in the progression of certain types of cancer. However, no comprehensive data is currently available on the expression of S100 family genes in lung adenocarcinomas. Oligonucleotide array, quantitative reverse transcription-polymerase chain reaction and western blot analyses of lung adenocarcinoma cell lines and bronchiolar epithelial cells (SAEC and NHBE) revealed that S100A2 and S100A4 were the most strikingly downregulated and upregulated members of the S100 family, respectively. Immunohistochemical analyses of 94 primary lung adenocarcinomas showed that positive S100A2 expression (33/94, 35.1%) was significantly associated with lymphatic invasion (P = 0.0233) and positive S100A4 expression (19/94, 20.2%) with vascular invasion (P=0.0454). Interestingly, a strong inverse relationship was found between S100A4 and p53 expression (P = 0.0008). Survival analyses showed that S100A4 positivity was associated with poor patient prognosis (P = 0.042). S100A2 positivity was not associated with patient survival when the whole patient group was analyzed; however, S100A2 positivity was a favorable prognostic indicator in patients with p53-negative tumors (P = 0.0448). Finally, we used oligonucleotide array analyses and identified potential S100A2 and S100A4 target genes involved in cancer progression: S100A2 induced RUNX3 and REPRIMO; S100A4 induced EZRIN, RUNX1 and WISP1; S100A2 repressed EGFR, NFKB2 and RELA2; and S100A4 repressed ANXA10 and IL1RN. Thus, the present study demonstrates involvement of S100A2 and S100A4 in the progression of lung adenocarcinomas and an inverse association between S100A4 and p53 expression, and provides a list of targets regulated by S100A2 and S100A4.
  • H Uozaki, JM Chong, E Fujimoto, M Itoh, M Saito, K Sakuma, M Sudo, T Ushiku, T Niki, H Nagai, K Takada, M Fukayama
    ANTICANCER RESEARCH 25 5 3183 - 3190 2005年09月 [査読有り][通常論文]
     
    Background: Epstein-Barr-virus-associated gastric carcinoma (EBVaGC) is a distinct subset of gastric carcinoma (GC). The expressions of cytokeratins (CK) 7, 8, 18, 19 and 20 and truncated basic hair keratin 1 (hHb1-AN) were investigated in GC to clarify the characteristics of EBVaGC. Materials and Methods: For immunohistochemical evaluation, 173 GC tissues were examined and 31 GC tissues and 5 GC cell lines were used for quantitative real-time RT-PCR (qrt-RT-PCR) and RT-PCR. Results: EBVaGC showed significantly lower immunohistochemical positivity of CK7 (-/+/-/++/+++; 27/15/4/1/1) compared to EBV-negative GC (12129127144113), even after stratification by histological types. The qrt-RT-PCR test demonstrated decreased amounts of CK7, 18 and 19 mRNAs in EBVaGC. Two among 5 GC cell lines showed a decrease of CK7 mRNA level after recombinant EBV infection. hHb1-Delta N expression was not specific to EBVaGC. Conclusion: Abnormalities of CK7, 18 and 19 expressions, especially a decreased amount of CK7 expression, are characteristics of EBV-associated epithelial malignancies and might be important in carcinogenesis.
  • A Goto, T Niki, CP Li, D Matsubara, Y Murakami, N Funata, M Fukayama
    CANCER SCIENCE 96 8 480 - 486 2005年08月 [査読有り][通常論文]
     
    The TSLC1 (tumor suppressor in lung cancer 1) gene is a tumor suppressor recently identified through functional complementation in a lung adenocarcinoma cell line A549. In this study we immunohistochemically examined the loss of TSLC1 expression in 93 cases of surgically resected lung adenocarcinoma, and investigated its correlation with clinicopathological parameters, including histological subtypes of tumors. The prognostic significance of loss of TSLC1 expression was analyzed by univariate and multivariate analyses, in parallel with other prognostic markers such as p53, p27, and Ki-67. In non-cancerous lung tissue, TSLC1 was weakly positive in bronchial and bronchiolar epithelial cells, type II pneumocytes and bronchial glands. Overall, TSLC1 was negative in 60 of 93 lung adenocarcinomas. TSLC1 was mainly localized in the cytoplasm of the cells, but cell membrane staining was also observed, especially at sites of cell-cell adhesion. TSLC1-negative tumors were more frequently observed in male cases (41/54 cases, 70.0%) than in female cases (19/39 cases, 48.7%) (P<0.01). Notably, TSLC1 expression was preserved in a non-invasive, bronchiolo-alveolar histological pattern of tumor cells (P<0.0001). Survival analyses showed that loss of TSLC1 expression was associated with lower patient survival in univariate and multivariate analyses (P<0.05 and P=0.059, respectively). Subset analyses further showed that the prognostic impact of loss of TSLC1 was significant for male patients (P=0.0089), but not for female patients. We conclude that TSLC1 is expressed in a subset of lung adenocarcinomas, especially in those with bronchiolo-alveolar spread pattern. Loss of TSLC1 is associated with lower patient survival, supporting its role as a tumor suppressor.
  • S Fukumoto, N Yamauchi, H Moriguchi, Y Hippo, A Watanabe, J Shibahara, H Taniguchi, S Ishikawa, H Ito, S Yamamoto, H Iwanari, M Hironaka, Y Ishikawa, T Niki, Y Sohara, T Kodama, M Nishimura, M Fukayama, H Dosaka-Akita, H Aburatani
    CLINICAL CANCER RESEARCH 11 5 1776 - 1785 2005年03月 [査読有り][通常論文]
     
    Purpose: Squamous cell carcinoma (SCC) and adenocarcinoma of the lung are currently subject to similar treatment regimens despite distinct differences in histology and epidemiology. The aim of this study is to identify a molecular target with diagnostic and therapeutic values for SCC. Experimental Design: Genes specifically up-regulated in SCC were explored through microarray analysis of 5 SCCs, 5 adenocarcinomas, 10 small cell lung carcinomas, 27 normal tissues, and 40 cancer cell lines. Clinical usefulness of these genes was subsequently examined mainly by immunohistochemical analysis. Results: Seven genes, including aldo-keto, reductase family 1, member B10 (AKR1B10), were identified as SCC-specific genes. AKR1B10 was further examined by immunohistochemical analysis of 101 non-small cell lung carcinomas (NSCLC) and its overexpression was observed in 27 of 32 (84.4%) SCCs and 19 of 65 (29.2%) adenocarcinomas. Multiple regression analysis showed that smoking was an independent variable responsible for AKR1B10 overexpression in NSCLCs (P < 0.01) and adenocarcinomas (P < 0.01). AKR1B10 staining was occasionally observed even in squamous metaplasia, a precancerous lesion of SCC. Conclusion: AKR1B10 was overexpressed in most cases with SCC, which is closely associated with smoking, and many adenocarcinoma cases of smokers. These results suggest that AKR1B10 is a potential diagnostic marker specific to smokers' NSCLCs and might be involved in tobacco-related carcinogenesis.
  • A Goto, J Nakajima, K Hara, T Niki, M Fukayama
    VIRCHOWS ARCHIV 446 1 73 - 77 2005年01月 [査読有り][通常論文]
     
    A 46-year-old man presented with a lung tumor 17 years after a subtotal colectomy and 13 years after a partial duodenectomy for familial adenomatous polyposis (FAP). There had been no malignant transformation in the specimens from his colectomy and duodenectomy, and a current gastrointestinal investigation revealed no evidence of malignancy. Pathological analysis of the lung tumor demonstrated adenocarcinoma with clear cells and a papillary structure, accompanied by tiny tumorous nodules in the background lung parenchyma. Many of the nodules were multifocal adenocarcinoma; however, some of the nodules demonstrated atypical adenomatous hyperplasia (AAH). This is the first case report of a lung adenocarcinoma accompanied by AAH in a FAP patient. Immunohistochemical and loss of heterozygosity studies revealed unique features of the lesions reflecting a disruption of the adenomatous poliposis coli - beta- catenin pathway.
  • A Goto, T Niki, S Moriyama, N Funata, H Moriyama, Y Nishimura, R Tsuchida, J Kato, M Fukayama
    PATHOLOGY INTERNATIONAL 54 9 675 - 681 2004年09月 [査読有り][通常論文]
     
    To clarify the association of the P27 degradation pathway proteins, Skp2 and Jab1, with the development and progression of lung adenocarcinoma (AD), we immunohistochemically investigated Skp2 and Jab1 expression together with P27- and Ki-67-labeling in 110 lung AD and 11 atypical adenomatous hyperplasia (AAH) and analyzed the relationship between the expression of these proteins and the clinicopathological factors. High Skp2 or Jab1 expression was frequent in lung AD (52/110, 47%, and 59/110, 54%, respectively), and high expression of Jab1 was also frequent in AAH (4/11, 36%), while it was not observed in normal bronchiolar epithelium. The P27 labeling index (LI) was reciprocally correlated with high Skp2 and Jab1 expression, and a higher Ki-67 LI was significantly correlated with high Skp2 and Jab1 expression. However, low P27 expression did not correlate with a higher Ki-67 LI. High Skp2 lung AD showed significant correlation with blood and lymphatic vessel invasion, which low P27 expression did not correlate with. Furthermore, high Skp2 expression in lung AD was significantly correlated with a poor outcome for patients. Thus, Skp2 and Jab1 regulate P27 degradation, and might contribute to the development and progression of lung AD through P27-mediated and -unmediated mechanisms.
  • J Shibahara, A Goto, T Niki, M Tanaka, A Nakajima, M Fukayama
    AMERICAN JOURNAL OF SURGICAL PATHOLOGY 28 6 825 - 829 2004年06月 [査読有り][通常論文]
     
    We describe a case of primary pulmonary paraganglioma, a tumor that has not been reported in sufficient detail in previous literature. The patient was a 55-year-old woman with hypertension accompanied by an elevated serum norepinephrine level (2651 pg/mL; normal 100-450 pg/mL). Computed tomography revealed a well-circumscribed solid mass, 3.5 cm in diameter, located in the lower lobe of the left hung. In the lobectomy specimen, the tumor had invaded the B8 bronchus and hilar lymph nodes with microscopic metastasis to the mediastinal nodes. The tumor showed histologic, immunohistochemical, and ultrastructural features of paraganglioma: argyrophilic cells arranged in a nesting (Zellballen) or anastomosing trabecular pattern within an arcuate vascular network. Neoplastic chief cells positive for neuroendocrine markers (CD56, synaptophysin, chromogranin A) were surrounded by sustentacular cells positive for S-100 protein. Neurofilament protein was positively stained, but cytokeratins were totally negative. On electron microscopy, chief cells possessed abundant dense core granules with an eccentric halo ("norepinephrine-type" granules). The patient's blood pressure began decline soon after the resection, and her serum norepinephrine promptly returned to almost normal. On the basis of our experience, our case is a bona fide primary pulmonary paraganglioma, a tumor heretofore subject to considerable skepticism.
  • H Kitagawa, A Goto, T Niki, M Hironaka, J Nakajima, M Fukayama
    PATHOLOGY INTERNATIONAL 53 12 823 - 827 2003年12月 [査読有り][通常論文]
     
    Atypical adenomatous hyperplasia (AAH) of the lung has been proposed as a possible precursor lesion of adenocarcinoma of the lung. In the present study, we sought to clarify the clinicopathological characteristics of lung adenocarcinoma cases associated with AAH, with special reference to tobacco smoking and the presence of multiple primary carcinomas of pulmonary and extrapulmonary organs. We examined 123 surgically resected lung adenocarcinomas and conducted histopathological diagnoses for AAH and multiple primary pulmonary carcinomas. Clinicopathological characteristics such as age, sex, smoking index, survival, and the presence of extrapulmonary primary carcinomas were obtained from clinical records, and the associations among these factors were examined statistically. Sixteen lung adenocarcinoma patients had accompanying AAH (the AAH group) and 107 cases did not (the NAAH group). The incidence of primary carcinomas in extrapulmonary organs was higher in the AAH group (37.5%; 6/16) than in the NAAH group (12.5%; 13/107) (P = 0.01). Multiple primary lung cancers tended to be more frequent in the AAH group, but the difference was not statistically significant (P = 0.07). Although there was no difference in tobacco smoking between the two groups, all eight cases with multiple primary lung carcinomas were smokers. Furthermore, multiple primary lung carcinomas were found more frequently in smokers of the AAH group (37.5%; 3/8) than in the smokers of the NAAH group (7.2%; 5/69) (P = 0.04). The results suggested that constitutional or genetic factors might predispose patients to the development of AAH together with extrapulmonary primary carcinomas, and that smoking might contribute to the development of multiple primary lung adenocarcinomas, especially in patients with pre-existing AAH.
  • H Terasaki, T Niki, Y Matsuno, T Yamada, A Maeshima, H Asamura, N Hayabuchi, S Hirohashi
    AMERICAN JOURNAL OF SURGICAL PATHOLOGY 27 7 937 - 951 2003年07月 [査読有り][通常論文]
     
    A significant proportion of small lung adenocarcinomas consists of two components: bronchioloalveolar carcinoma (BAC) and invasive carcinoma. The purpose of this study was to compare their clinicopathologic features with those of BAC and those of invasive cancer without BAC, and to define "early invasive" lesions based on the extent of invasive foci. We reviewed 484 lesions of resected lung adenocarcinoma and classified them into three groups according to tumor growth pattern: group 1 (n = 102, BAC), group 2 (n = 216, adenocarcinoma consisting of BAC and invasive carcinoma), and group 3 (n = 166, invasive adenocarcinoma without BAC component). Group 2 was further subdivided according to the extent of the invasive area: group 2a (n = 54), BAC with invasive foci less than or equal to5 mm; group 2b (n = 162), BAC with invasive foci >5 mm. These groups were compared with regard to their clinicopathologic features, expression of Ki-67 and p53, and expression of laminin-5, a putative marker for tumor invasion. The positivity rates of vascular, lymphatic, and pleural invasion in each group were as follows: 0%, 0%, and 0% in group 1; 5.5%, 14.8%, and 1.9% in group 2a; 45.7%, 41.4%, and 25.9% in group 2b; and 84.9%, 61.4%, and 60.8% in group 3. Notably, no lymph node metastasis occurred in either group 2a or group 1, but it was observed in 24.1% of group 2b and 47.0% of group 3. The mean Ki-67 labeling index, the frequency of p53 overexpression, and the frequency of laminin-5 overexpression increased from group 1 (11%, 4%, and 0%) to group 2a (16%, 20%, and 7%) to group 2b (24%, 41%, and 23%) to group 3 (35%, 38%, and 38%). In contrast, no clear differences were observed when lesions were subdivided according to size. Based on the distribution pattern of Ki-67-positive tumor cells, lesions were classified into two groups: marginal type (63%) and nonmarginal type (37%). The latter showed a significantly higher labeling index than the former. Moreover, the proportion of the marginal type clearly decreased from group 1 (85%) and group 2a (87%) to group 2b (55%) to group 3 (19%). Group 2 lesions showed characteristics intermediate between the BAC and invasive adenocarcinoma. According to the extent of the invasive area, we were able to define a subgroup of mixed-type adenocarcinomas (group 2a) that could be regarded as early invasive cancer because they showed low rates of vascular, lymphatic, and pleural invasion, and no nodal involvement.
  • H Kitagawa, A Goto, M Minami, J Nakajima, T Niki, M Fukayama
    JAPANESE JOURNAL OF CLINICAL ONCOLOGY 33 7 360 - 363 2003年07月 [査読有り][通常論文]
     
    A case of cystic sclerosing hemangioma of the lung is reported. A chest X-ray of a 55-year-old woman who had been suffering from a cough with sputum for several months revealed an abnormal nodular shadow. A chest CT scan revealed a solitary tumor with cystic appearance occupying S7 of the right lung and the inferior pulmonary ligament. Radiological differential diagnosis for the lesion included bronchogenic cyst, cystic Schwannoma, pulmonary necrotic carcinoid, and lung carcinoma. Right lower lobectomy was carried out and the lesion was pathologically diagnosed as sclerosing hemangioma of the lung with cystic features, expanding into the pulmonary ligament. Differential diagnosis of the cystic lesion of the lung should include cystic sclerosing hemangioma as observed in this case.
  • AM Maeshima, T Niki, A Maeshima, T Yamada, H Kondo, Y Matsuno
    CANCER 95 12 2546 - 2554 2002年12月 [査読無し][通常論文]
     
    BACKGROUND. Several studies have shown the prognostic value of desmoplasia for lung adenocarcinomas. The authors evaluated the density and extent of desmoplasia by modifying the scar grade, as well as the prognostic impact on patient survival. METHODS. Modified scar grade was defined as follows: Grade 1, no desmoplasia; Grade 2, sparse desmoplastic reaction; Grade 3, dense desmoplastic reaction with diameter of 10 mm or less; Grade 4, dense desmoplastic reaction with diameter exceeding 10 turn. In addition, the prognostic impact of conventional histologic factors and modified scar grade was analyzed in 239 cases of small peripheral lung adenocarcinoma (maximum dimension, ! 30 mm) for which long-term follow-up data were available. RESULTS. The 5 and 10-year survival rates according to the modified scar grade were 100% and 100% for Grade I lung adenocarcinoma (n = 29); 91.7% and 83.7% for Grade 2 (n 61); 67.6% and 52.7% for Grade 3 (n = 78); and 50.0% and 37.5% for Grade 4 (n 71), respectively. A significant difference in patient survival was found between Grade I or 2 versus Grade 3 or 4 (P < 0.0001, by log rank test). Multivariate analysis showed that modified scar grade was an independent prognostic factor (P = 0.0176), as were pathologic stage (P = 0.0293), lymph node metastasis (P = 0.0191), lymphatic permeation (P = 0.0022), and pleural involvement (P = 0.0452). Modified scar grade also had a significant impact on survival in various subsets of patients, including those with pathologic Stage IA disease, patients with tumors of diameter 20 mm or less, or patients with mixed subtype tumors with a bronchioloalveolar component. CONCLUSIONS. Modified scar grade is a useful prognostic factor in patients with small lung adenocarcinomas. Tumors with a sparse fibroblastic reaction (modified scar Grade 2) may represent early invasive cancers or invasive cancers with low malignant potential, which should be distinguished from frankly invasive cancers (modified scar Grade 3 or 4). Cancer 2002;95:2546-54. (C) 2002 American Cancer Society.
  • AM Maeshima, T Niki, A Maeshima, T Yamada, H Kondo, Y Matsuno
    CANCER 95 12 2546 - 2554 2002年12月 [査読有り][通常論文]
     
    BACKGROUND. Several studies have shown the prognostic value of desmoplasia for lung adenocarcinomas. The authors evaluated the density and extent of desmoplasia by modifying the scar grade, as well as the prognostic impact on patient survival. METHODS. Modified scar grade was defined as follows: Grade 1, no desmoplasia; Grade 2, sparse desmoplastic reaction; Grade 3, dense desmoplastic reaction with diameter of 10 mm or less; Grade 4, dense desmoplastic reaction with diameter exceeding 10 turn. In addition, the prognostic impact of conventional histologic factors and modified scar grade was analyzed in 239 cases of small peripheral lung adenocarcinoma (maximum dimension, ! 30 mm) for which long-term follow-up data were available. RESULTS. The 5 and 10-year survival rates according to the modified scar grade were 100% and 100% for Grade I lung adenocarcinoma (n = 29); 91.7% and 83.7% for Grade 2 (n 61); 67.6% and 52.7% for Grade 3 (n = 78); and 50.0% and 37.5% for Grade 4 (n 71), respectively. A significant difference in patient survival was found between Grade I or 2 versus Grade 3 or 4 (P < 0.0001, by log rank test). Multivariate analysis showed that modified scar grade was an independent prognostic factor (P = 0.0176), as were pathologic stage (P = 0.0293), lymph node metastasis (P = 0.0191), lymphatic permeation (P = 0.0022), and pleural involvement (P = 0.0452). Modified scar grade also had a significant impact on survival in various subsets of patients, including those with pathologic Stage IA disease, patients with tumors of diameter 20 mm or less, or patients with mixed subtype tumors with a bronchioloalveolar component. CONCLUSIONS. Modified scar grade is a useful prognostic factor in patients with small lung adenocarcinomas. Tumors with a sparse fibroblastic reaction (modified scar Grade 2) may represent early invasive cancers or invasive cancers with low malignant potential, which should be distinguished from frankly invasive cancers (modified scar Grade 3 or 4). Cancer 2002;95:2546-54. (C) 2002 American Cancer Society.
  • M Nishioka, T Kohno, M Tani, N Yanaihara, Y Tomizawa, A Otsuka, S Sasaki, K Kobayashi, T Niki, A Maeshima, Y Sekido, JD Minna, S Sone, J Yokota
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 99 19 12269 - 12274 2002年09月 [査読有り][通常論文]
     
    Loss of heterozygosity on chromosome 22q has been detected in approximately 60% of advanced nonsmall cell lung carcinoma (NSCLC) as well as small cell lung carcinoma (SCLC), suggesting the presence of a tumor suppressor gene on 22q that is involved in lung cancer progression. Here, we isolated a myosin family gene, MYO18B, located at chromosome 22q12.1 and found that it is frequently deleted, mutated, and hypermethylated in lung cancers. Somatic MYO18B mutations were detected in 19% (14/75) of lung cancer cell lines and 13% (6/46) of primary lung cancers of both SCLC and NSCLC types. MYO18B expression was reduced in 88% (30/34) of NSCLC and 47% (8/17) of SCLC cell lines. Its expression was restored by treatment with 5-aza-2'-deoxycytidine in 11 of 14 cell lines with reduced MYO18B expression, and the promoter CpG island of the MYO18B gene was methylated in 17% (8/47) of lung cancer cell lines and 35% (14/40) of primary lung cancers. Furthermore, restoration of MYO18B expression in lung carcinoma cells suppressed anchorage-independent growth. These results indicate that the MYO18B gene is a strong candidate for a novel tumor suppressor gene whose inactivation is involved in lung cancer progression.
  • K Rombouts, T Niki, P Greenwel, A Vandermonde, A Wielant, K Hellemans, P De Bleser, M Yoshida, D Schuppan, M Rojkind, A Geerts
    EXPERIMENTAL CELL RESEARCH 278 2 184 - 197 2002年08月 [査読無し][通常論文]
     
    Excessive production of collagens by a-smooth muscle actin (alpha-SMA)-positive myofibroblasts leads to fibrotic skin diseases, such as hypertrophic scarring. This process is characterized by an imbalance between extracellular matrix (ECM) synthesis and degradation, while transforming growth factor beta (TGF-beta(1)), known to be a key mediator of fibrogenesis, is up-regulated. In this study we have investigated the possible antifibrogenic effect of Trichostatin A (TSA), a histone deacetylase inhibitor, on rat skin fibroblasts in culture. mRNA steady-state levels and de novo protein synthesis of procollagen types I and III and alpha-SMA were inhibited when skin fibroblasts were treated with 100 nM TSA with or without TGF-beta(1). While the transcription rate of the procollagen alpha1(I) gene was increased following TSA or TGF-beta(1) treatment, TSA abrogated the stimulatory effect of TGF-beta(1) on procollagen alpha1(I) transcription when both compounds were added simultaneously. The reduction of procollagen alpha1(I) and alpha1(III) mRNA steady-state levels by TSA did not require de novo protein synthesis; while the effect of TSA on alpha-SMA mRNA steady-state levels was cycloheximide-sensitive. Interestingly, TSA affected TGF-beta(1) and its downstream mediators, i.e., the Smad family proteins. TSA strongly induced in a biphasic way the expression of 5'TG3' interacting factor (TGIF), a known endogenous corepressor molecule of the TGF-beta(1) signaling pathway. Addition of exogenous TGF-beta(1) did not interfere with the effect of TSA on the TGIF mRNA level. Our study shows that inhibition of histone deacetylases by TSA reduces expression of fibrosis-related genes in skin fibroblasts and this coincides by alterations in the TGF-beta(1) signaling pathway. (C) 2002 Elsevier Science (USA).
  • H Takei, H Asamura, A Maeshima, K Suzuki, H Kondo, T Niki, T Yamada, R Tsuchiya, Y Matsuno
    JOURNAL OF THORACIC AND CARDIOVASCULAR SURGERY 124 2 285 - 292 2002年08月 [査読有り][通常論文]
     
    Objective: Large cell neuroendocrine carcinoma of the lung is a newly recognized clinicopathologic entity. The clinical characteristics and optimal treatment of patients with large cell carcinomas are not yet established. The aim of this study was to define the clinicopathologic characteristics of large cell neuroendocrine carcinoma. Methods: The histologic characteristics of the patients receiving an initial diagnosis of poorly differentiated non-small cell lung carcinoma (n = 484), small cell carcinoma (n = 55), carcinoid (n = 31), and large cell neuroendocrine carcinoma (n = 12) were retrospectively reviewed according to World Health Organization criteria. Immunohistochemistry was performed to confirm the neuroendocrine phenotype. The outcomes and other clinical characteristics of those patients with large cell neuroendocrine carcinoma were retrospectively analyzed and compared with those of patients with poorly differentiated carcinoma of other histologic types. Results: A total of 87 patients were given a diagnosis of large cell neuroendocrine carcinoma after the histologic review. These patients comprised 3.1% of all patients undergoing resection for primary lung cancer during the same period. The overall 5-year survival was 57%. The 5-year survivals of patients with stage I, II, III, and IV disease were 67%. 75%, 45%, and 0%, respectively. There was no statistically significant difference between the overall survival of patients with large cell neuroendocrine carcinoma and those with other non-small cell lung cancers. There was a significant difference between the survival of patients with stage I large cell neuroendocrine carcinoma and that of patients with the same stage of other non-small cell lung carcinomas. The site of the first documented recurrence was locoregional in 12 patients (34%), distant metastases in 20 patients (57%), and both simultaneously in 3 patients. Locoregional lymph node recurrences were observed frequently. More than 80%. of recurrences were found within 1 year after the operation. Conclusion: in terms of prognosis, large cell neuroendocrine carcinoma is distinctly different from other non-small cell lung cancers. The prognosis of large cell neuroendocrine carcinoma was poor, even for early stage disease; the prognosis of the stage I disease of large cell neuroendocrine carcinoma was poorer than that of the same stage of other non-small cell lung cancers. Because of its aggressive clinical behavior and poor prognosis, large cell neuroendocrine carcinoma should be recognized as one of the poorest prognostic subgroups among primary lung cancers, and therefore novel therapeutic approaches should be established.
  • K Rombouts, T Niki, P Greenwel, A Vandermonde, A Wielant, K Hellemans, P De Bleser, M Yoshida, D Schuppan, M Rojkind, A Geerts
    EXPERIMENTAL CELL RESEARCH 278 2 184 - 197 2002年08月 [査読有り][通常論文]
     
    Excessive production of collagens by a-smooth muscle actin (alpha-SMA)-positive myofibroblasts leads to fibrotic skin diseases, such as hypertrophic scarring. This process is characterized by an imbalance between extracellular matrix (ECM) synthesis and degradation, while transforming growth factor beta (TGF-beta(1)), known to be a key mediator of fibrogenesis, is up-regulated. In this study we have investigated the possible antifibrogenic effect of Trichostatin A (TSA), a histone deacetylase inhibitor, on rat skin fibroblasts in culture. mRNA steady-state levels and de novo protein synthesis of procollagen types I and III and alpha-SMA were inhibited when skin fibroblasts were treated with 100 nM TSA with or without TGF-beta(1). While the transcription rate of the procollagen alpha1(I) gene was increased following TSA or TGF-beta(1) treatment, TSA abrogated the stimulatory effect of TGF-beta(1) on procollagen alpha1(I) transcription when both compounds were added simultaneously. The reduction of procollagen alpha1(I) and alpha1(III) mRNA steady-state levels by TSA did not require de novo protein synthesis; while the effect of TSA on alpha-SMA mRNA steady-state levels was cycloheximide-sensitive. Interestingly, TSA affected TGF-beta(1) and its downstream mediators, i.e., the Smad family proteins. TSA strongly induced in a biphasic way the expression of 5'TG3' interacting factor (TGIF), a known endogenous corepressor molecule of the TGF-beta(1) signaling pathway. Addition of exogenous TGF-beta(1) did not interfere with the effect of TSA on the TGIF mRNA level. Our study shows that inhibition of histone deacetylases by TSA reduces expression of fibrosis-related genes in skin fibroblasts and this coincides by alterations in the TGF-beta(1) signaling pathway. (C) 2002 Elsevier Science (USA).
  • Y Tomizawa, T Kohno, H Kondo, A Otsuka, M Nishioka, T Niki, T Yamada, A Maeshima, K Yoshimura, R Saito, JD Minna, J Yokota
    CLINICAL CANCER RESEARCH 8 7 2362 - 2368 2002年07月 [査読有り][通常論文]
     
    Purpose: Chromosome 3p is deleted frequently in various types of human cancers, including lung cancer. Recently, the RASSF1A gene was isolated from the 3p21.3 region homozygously deleted in lung and breast cancer cell lines, and it was shown to be inactivated by hypermethylation of the promoter region in lung cancers. In this study, we investigated the pathogenetic and clinicopathological significances of RASSF1A methylation in the development and/or progression of lung adenocarcinoma. Experimental Design: Association of RASSF1A methylation with clinicopathological features, allelic imbalance at 3p21.3, p53 mutations, and K-ras mutations was examined in 110 stage 1 lung adenocarcinomas. Results: Thirty-five of 110 (32%) tumors showed RASSF1A methylation. RASSF1A methylation was dominantly detected in tumors with vascular invasion (P = 0.0242) or pleural involvement (P = 0.0305), and was observed more frequently in poorly differentiated tumors than in well (P = 0.0005) or moderately (P = 0.0835) differentiated tumors. Furthermore, RASSF1A methylation correlated with adverse survival by univariate analysis (P = 0.0368; log-rank test) as well as multivariate analysis (P = 0.032,; risk ratio 2.357; 95% confidence interval, 1.075-5.169). The correlation between RASSF1A methylation and allelic imbalance at 3p21.3 was significant (P = 0.0005), whereas the correlation between RASSF1A methylation and p53 mutation was borderline (P = 0.0842). However, there was no correlation or inverse correlation between RASSF1A methylation and K-ras mutation (P = 0.2193). Conclusions: These results indicated that epigenetic inactivation of RASSF1A plays an important role in the progression of lung adenocarcinoma, and that RASSF1A hypermethylation appears to be a useful molecular marker for the prognosis of patients with stage I lung adenocarcinoma.
  • T Niki, T Kohno, S Iba, Y Moriya, Y Takahashi, M Saito, A Maeshima, T Yamada, Y Matsuno, M Fukayama, J Yokota, S Hirohashi
    AMERICAN JOURNAL OF PATHOLOGY 160 3 1129 - 1141 2002年03月 [査読有り][通常論文]
     
    Laminin-5 is an extracellular matrix protein that plays a key role in cell migration and tumor invasion. Cox-2 is an induced isoform of cyclooxygenases that plays an important role in carcinogenesis, suppression of apoptosis, angiogenesis, and metastasis of colon cancer. We report frequent co-expression of cox-2 and laminin-5 at the invasive front of earlystage lung adenocarcinomas. We investigated the expression of cox-2 and laminin-5 immunohistochemically in 102 cases of small-sized lung adenocarcinoma (maximum dimension, 2 cm or less). Cox-2 and laminin-5 were expressed in 97 (95.1%) and 82 (80.4%) cases, respectively. Both were preferentially localized in cancer cells at the cancer-stroma interface, although cox-2 tended to show a diffuse staining pattern in some cases. A comparison of their staining patterns revealed a striking similarity in their distribution in 24 cases, and a partial overlap between their localization in another 20 cases. Moreover, an overall correlation was found between the expression levels of cox-2 and laminin-5 (P = 0.018). To gain insight into the mechanisms that regulate the expression of these proteins, we additionally studied their expression in 58 cases of stage I lung adenocarcinoma, in which P53 status was determined by immunohistochemistry, polymerase chain reaction-single strand conformation polymorphism analysis, and direct sequencing. The results showed that tumors with mutant p53 tended to express more cox-2 than those with wild-type p53 (P = 0.080). Also, tumors that overexpressed P53 had higher levels of cox-2 and laminin-5 than those without P53 overexpression (P = 0.032 and 0.047, respectively). Further immunohistochemical analysis showed that tumors that over-expressed both epidermal growth factor receptor (EGFR) and erbB-2 had higher levels of cox-2 and laminin-5 than those without concomitant overexpression of these proteins (P = 0.014 and P = 0.018, respectively). To see whether EGFR signaling is involved in cox-2 and laminin-5 expression, we further conducted in vitro analyses using six lung adenocarcinoma cell lines (A549, HLC-1, ABC-1, LC-2/ad, VMRC-LCD, and L27). Western blot analyses showed that cox-2 mRNA levels, and to a lesser extent laminin-5 gamma(2) mRNA levels, correlated with the expression levels of erbB-2 and the phosphorylated form of MAPK/ERK-1/2 protein. The addition of transforming growth factor-alpha increased both cox-2 and laminin-5 gamma2 n3RNA levels in A549, ABC-1, and L27 with different kinetics; the induction of cox-2 occurred earlier th an that of laminin-5 gamma2. Finally, the migration of ABC-1 cells was inhibited by MAP kinase kinase inhibitor PD98059 and a selective cox-2 inhibitor NS-398. In contrast, the migration of A549 cells was inhibited by PD98059, but much less effectively by NS-398. These results suggest that co-stimulatory mechanisms may exist that increase the expression of cox-2 and laminin-5 at the invasive front of lung adenocarcinomas and that EGFR signaling could be one of the mechanisms. Further investigations are warranted concerning the role of cox-2 and laminin-5 in cancer cell invasion and the significance of p53 and EGFR signaling in the regulation of cox-2 and laminin-5 expression.
  • Frequent Co-Localization of Cox-2 and Laminin-s gamma2 Chain at the Invasive Front of Early-Stage Lung Adenocarcinomas.
    Niki T, Kohno T, Iba S, Moriya Y, Takahashi Y, Saito M, Maeshima A, Yamada T, Matsuno Y, Fukayama M, Yokota J, Hirohashi S
    Am J Pathol 160 1129 - 1141 2002年 [査読無し][通常論文]
  • Clinicopathological significance of epigenetic inactivation of RASSFIA at 3p2l.3 in stage I lung adenocarcinoma.
    Tomizawa Y, Kohno T, Kondo H, Otsuka A, Nishioka M, Niki T, Yamada T, Maeshima A, Yoshimura K, Saito R, Minna JD, Yokota J
    Crin Cancer Res. 8 2362 - 2368 2002年 [査読無し][通常論文]
  • Nishioka M, Kohno T, Tani M, Yanaihara N, Tomizawa Y, Otsuka A, Sasaki S, Kobayashi K, Niki T, Maeshima A, Sekido Y, Minna JD, Sone S, Yokota J
    Proc Natl Acad Sci U S A. 99 12269 - 12274 2002年 [査読無し][通常論文]
  • M Tokunou, T Niki, K Eguchi, S Iba, H Tsuda, T Yamada, Y Matsuno, H Kondo, Y Saitoh, H Imamura, S Hirohashi
    AMERICAN JOURNAL OF PATHOLOGY 158 4 1451 - 1463 2001年04月 [査読無し][通常論文]
     
    Hepatocyte growth factor (HGF) plays important rots in tumor development and progression. It is currently thought that the main action of HGF is of a paracrine nature: HGF produced by mesenchymal cells acts on epithelial cells that express its receptor c-MET. In this investigation, we explored the significance of c-MET expression in myofibroblasts, both in culture and in patients with lung adenocarcinoma. We first showed that human myofibroblasts derived from primary lung cancer expressed c-MET mRNA and protein by reverse transcription-polymerase chain reaction and Western blot analysis. Proliferation of myofibroblasts was stimulated in a dose-dependent manner by exogenously added recombinant human HGF whereas it was inhibited in a dose-dependent manner by neutralizing antibody to HGF. The addition of HGF in the culture medium stimulated tyrosine phosphorylation of c-MET. The c-MET protein was immunohistochemically detected in myofibroblasts in the invasive area of lung adenocarcinoma. Finally, the prognostic significance of c-MET expression in stromal myofibroblasts was explored in patients with small-sized lung adenocarcinomas. c-MET-positive myofibroblasts were observed in 69 of 131 cases (53%). A significant relationship between myofibroblast c-MET expression and shortened patient survival was observed in a whole cohort of patients including ah pathological stages (two-sided P = 0.0089 by log-rank test) and in patients with stage IA disease (two-sided P = 0.0019 by log-rank test). These data suggest that the HGF/c-MET system constitutes an autocrine activation loop in cancer-stromal myofibroblasts. This autocrine system may play a role in invasion and metastasis of lung adenocarcinoma.
  • T Niki, S Iba, T Yamada, Y Matsuno, B Enholm, S Hirohashi
    JOURNAL OF PATHOLOGY 193 4 450 - 457 2001年04月 [査読無し][通常論文]
     
    Vascular endothelial growth factor receptor 3 (VEGFR-3) has been proposed as a marker for lymphatic endothelial cells. This study investigated the expression of VEGFR-3 in the tumour vessels of lung adenocarcinoma and evaluated whether VEGFR-3 staining was useful for identifying lymphatic vessels within the tumo. It also explored whether active growth of lymphatic vessels occurred in lung adenocarcinoma. Formalin-fixed, paraffin-embedded specimens obtained from 60 cases of lung adenocarcinoma, including five cases of pure bronchiolo-alveolar carcinoma (BAC) without stromal, vascular, and pleural invasion, were examined. No VEGFR-3-positive vessels were observed in pure BAC, but varying numbers of VEGFR-3-positive vessels were found in 39 of 55 (70.9%) invasive adenocarcinomas. A comparison of serial sections stained for VEGFR-3, CD31, and laminin-1 showed that most of the VEGFR-3-positive vessels appeared to be blood vessels (CD31-positive, laminin-1-positive), but some had the characteristics of lymphatic vessels (variable staining for CD31, little or no staining for laminin-1). VEGFR-3 staining highlighted lymphatic invasion by cancer tells; this invasion could not be detected by CD31 or haematoxylin and eosin (H&E) staining. Active growth of lymphatic vessels (as indicated by nuclear Ki-67 labelling of the endothelium) was observed in five tumours, four of which showed a high level of lymphatic invasion by cancer cells. It was concluded that VEGFR-3 immunostaining did not discriminate clearly between vascular and lymphatic endothelial cells, since expression of VEGFR-3 can be up-regulated in tumour blood vessels. However, VEGFR-3 staining combined with laminin-1 and CD31 staining would be useful for identifying lymphatic vessels and their invasion by tumour cells in a more objective way. Finally, proliferation of lymphatic endothelial cells may occur in association with lymphatic invasion by cancer cells. Copyright (C) 2001 John Wiley & Sons, Ltd.
  • Rombouts K, Niki T, Wielant A, Hellemans K, Schuppan D, Kormoss N, Geerts A
    J Hepatol 34 230 - 8 2001年 [査読無し][通常論文]
  • A Geerts, C Eliasson, T Niki, A Wielant, F Vaeyens, M Pekny
    HEPATOLOGY 33 1 177 - 188 2001年01月 [査読無し][通常論文]
     
    Increased desmin synthesis and formation of desmin-containing intermediate filaments (IFs) is one of the hallmarks of transdifferentiation of hepatic stellate cells into myofibroblast-like cells. These desmin-enriched myofibroblast-like cells are the major sources of fibrotic extracellular matrix in chronically diseased liver. Myofibroblast-like cells are also involved in the contraction of sinusoids, which leads to increased intrahepatic pressure and portal hypertension. To address the requirements for the formation of desmin-containing IFs both in quiescent and in transdifferentiated stellate cells, we used mice deficient for glial fibrillary acidic protein (GFAP) and/or vimentin, which are additional IF proteins present in stellate cells. In this study, we show that desmin cannot form full-length bundles of IFs in the absence of both GFAP and vimentin. Quiescent and transdifferentiated GFAP(-/-)vim(-/-) stellate cells are devoid of normal bundles of IFs. Instead, they exhibit only residual IF bundles restricted to subcortical cytoplasm, although these cells contain equal desmin mRNA and protein levels as wild-type cells. The absence of vimentin alone restricts formation of desmin-containing IF bundles to the perinuclear region, while both the distal processes in quiescent stellate cells and the subcortical zone in myofibroblast-like cells remain free of desmin-containing IF bundles. The absence of GFAP alone does not interfere with the formation of desmin-containing IFs. Thus, to form normal IFs in stellate cells, desmin is required to partnerize with vimentin. In addition, these mouse models will prove to be instrumental in addressing the role of IFs in the process of stellate cell transdifferentiation.
  • Moriya Y, Niki T, Yamada T, Matsuno Y, Kondo H, Hirohashi S
    Cancer 91 1129 - 1141 2001年 [査読無し][通常論文]
  • M Nishioka, T Kohno, M Takahashi, T Niki, T Yamada, S Sone, J Yokota
    ONCOGENE 19 54 6251 - 6260 2000年12月 [査読無し][通常論文]
     
    Frequent allelic losses on chromosome 22q in small cell lung carcinomas (SCLCs) and advanced non-small cell lung carcinomas indicate the presence of tumor suppressor gene(s) on this chromosome arm. We detected a homozygous deletion at 22q12.1 in a SCLC cell line, Lu24. Cloning of the breakpoints of the Lu24 deletion revealed that the deletion was interstitial and 428, 131 bp in size. The deleted region contained the SEZ6L (Seizure 6-like) gene, whose structure had been partially determined by the chromosome 22 sequencing project, We determined the full length cDNA sequence for the SEZ6L gene based on the genomic sequence for the SEZ6L locus using the GENSCAN program and the RT-PCR method. The deduced SEZ6L protein was a transmembrane protein of 1024 amino acids with multiple domains involved in protein-protein interaction and signal transduction, SEZ6L expression was detected in a variety of human tissues, including lung, while its expression was detected in 14 (30%) of 46 lung cancer cell lines examined, Missense mutations were detected in three (7%) of the 46 cell lines, and a 1 bp deletion in the polypyrimidine tract preceding exon 4 was detected in one (2%) of 46 primary lung cancers. Therefore, it is possible that genetic and/or epigenetic SEZ6L alterations are involved in the development and/or progression in a subset of lung cancer, although functional analysis of the SEZ6L gene as well as molecular analysis of other genes in the homozygously deleted region is necessary to understand the pathogenetic significance of 220 deletions in human lung carcinogenesis.
  • M Tokunou, T Niki, Y Saitoh, H Imamura, M Sakamoto, S Hirohashi
    LABORATORY INVESTIGATION 80 11 1643 - 1650 2000年11月 [査読無し][通常論文]
     
    Radixin is a member of the ERM (ezrin/radixin/moesin) protein family that is proposed to function as a membrane-cytoskeletal linker. Using differential display analysis, we have identified radixin as a gene down-regulated in primary lung adenocarcinoma. Real-time quantitative reverse transcription polymerase chain reaction confirmed that radixin mRNA was decreased, both in 10 early-stage bronchioloalveolar carcinomas and in 16 invasive lung adenocarcinomas, by 69% (p = 0.0002) and 82% (p < 0.0001), respectively, compared with 9 nontumor lung tissues. Similarly, moesin and ezrin mRNA levels were reduced in lung adenocarcinoma. Immunohistochemistry confirmed that cancer cells expressed very little radixin and moesin, whereas non-neoplastic alveolar and bronchiolar epithelial cells, and endothelial cells, including those within the tumor stroma, were consistently positive for these two proteins. Ezrin was localized in the apical surface of non-neoplastic bronchiolar and alveolar epithelial cells and, in contrast to radixin and moesin, the majority of tumor cells retained expression of ezrin. Localization of ezrin was altered in a significant proportion of tumor cells: whereas tumor cells forming lumina displayed membranous staining on the apical side, tumor cells with disorganized structures were either negative or diffusely positive for ezrin in the cytoplasm. Furthermore, a fraction of tumor cells invading the stroma in a scattered manner were strongly positive for ezrin. In conclusion, expression of radixin and moesin is down-regulated in lung adenocarcinoma, including early-stage bronchioloalveolar carcinoma. An intriguing implication of this finding is that these two genes may function as tumor suppressors in lung adenocarcinoma oncogenesis. Although structurally related to radixin and moesin, ezrin may have a distinct function in tumor-cell invasion.
  • R Manda, T Kohno, T Niki, T Yamada, S Takenoshita, H Kuwano, J Yokota
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 275 2 440 - 445 2000年08月 [査読無し][通常論文]
     
    To identify genes differentially expressed between small cell lung carcinoma (SCLC) cells and non-SCLC cells, mRNA differential display was applied to 3 SCLC cell lines and 6 non-SCLC cell lines. The LAMBS gene was identified as being expressed only in non-SCLC cells and not in SCLC cells. The LAMB3 gene encodes the laminin beta 3 chain, which is a unique component of laminin-5. Laminin-5 is a heterotrimer protein consisting of the alpha 3, beta 3, and gamma 2 chains, and another unique component of laminin-5 is the gamma 2 chain encoded by the LAMC2 gene. RT-PCR analysis of the LAMBS and LAMC2 genes in 45 lung cancer cell lines revealed that both the LAMB3 and LAMC2 genes were co-expressed in 21 of 32 non-SCLC cell lines (66%) but only in one of 13 SCLC cell lines (8%). Coexpression of the LAMB3 and LAMC2 genes was also observed in all 4 cases of primary non-SCLC cells examined but not in the corresponding non-cancerous lung cells. Since alpha 6 beta 4 integrin, the specific laminin-5 binding receptor, is known to be expressed only in non-SCLC cells and not in SCLC cells, it was indicated that laminin-5 is a critical microenvironmental factor for the growth of non-SCLC cells but not of SCLC cells, The differences in the expression of integrins and laminins would be critical factors to distinguish SCLC and non-SCLC cells, and such differences might be associated with the unique biological properties of SCLC cells, including metastatic potential and drug sensitivity. (C) 2000 Academic Press.
  • T Niki, S Iba, M Tokunou, T Yamada, Y Matsuno, S Hirohashi
    CLINICAL CANCER RESEARCH 6 6 2431 - 2439 2000年06月 [査読無し][通常論文]
     
    Vascular endothelial growth factors (VEGFs) C and D are novel members of the VEGF family that show some selectivity toward lymphatic endothelial cells. Recent studies suggest that VEGF-C may be involved in lymphangiogenesis and spread of cancer cells via lymphatic vessels. However, whether other VEGF family members play a role in lymph node metastasis is largely unknown. The aim of the present study was to explore whether expressions of VEGF-A, VEGF-B, VF,GF-C, and VEGF-D are correlated with lymph node status in lung adenocarcinoma. Total RNA was isolated from 60 surgical specimens of lung adenocarcinoma with (n = 27) or without (n = 33) lymph node metastasis. The relative mRNA abundance of VEGF-A, VEGF-B, VEGF-C, and VEGF-D was measured by real-time reverse transcription-PCR analysis based on TaqMan fluorescence methodology. We found that, as single factors, expression of none of the four VEGF family members clearly correlated with the presence of lymph node metastasis. The only tendency noted was for higher VEGF-B and VEGF-C and lower VEGF-D levels in the node-positive group. However, two-way scatterplot analysis revealed that tumors with lymph node metastasis were associated with a pattern of low VEGF-D and high VEGF A, VEGF-B, or VEGF-C, such that the ratios of VEGF-D:VEGF-A, VEGP-D:VEGF-B, or VEGF-D:VEGF-C were significantly lower in the node-positive group. Strikingly, none of the 11 tumors with high VEGP-D levels metastasized to lymph nodes. Furthermore, a low VEGF-D:VEGF-C ratio correlated with the presence of lymphatic invasion, and six of seven tumors with a pattern of very high expression of VEGF-C and low expression of VEGF-D displayed lymph vessel invasion that extended along the bronchovascular tree beyond the main tumor. Finally, levels of VEGF-A, but not VEGF-B or VEGF-C, were higher in tumors with large nodal metastasis (greater than or equal to 1 cm) than in those with small (<1 cm) nodal metastasis. These results support the hypothesis that two VEGF family members are involved in lymph node metastasis at two distinct steps; VEGF-C facilitates entry of cancer cells into the lymph vasculature, whereas VEGF-A promotes the growth of metastatic tumor through angiogenesis. The results also suggest that the balance between VEGF-C and VEGF-D could be important rather than the level of VEGF-C alone. Whether a low VEGF-D level plays a causative role in lymph node metastasis requires further investigation.
  • Y Ono, Y Nakanishi, Y Ino, T Niki, T Yamada, K Yoshimura, M Saikawa, T Nakajima, S Hirohashi
    CANCER 85 11 2315 - 2321 1999年06月 [査読無し][通常論文]
     
    BACKGROUND, The laminin-5 gamma 2 chain plays an important role in cell migration during tumor invasion and tissue remodeling. METHODS. Laminin-5 gamma 2 chain expression in squamous cell carcinomas of the tongue in 67 patients with Stage II, III, or IVA,B (excluding the cases with distant metastasis) was examined immunohistochemically to determine its associations with the clinicopathologic features of each tumor. The predominant staining patterns were categorized as follows: A, few or no tumor cells were positive; B, part of the tumor nest periphery was positive; C, the tumor nest periphery was circumferentially positive; or D, almost all the tumor cells were positive. RESULTS. Laminin-5 gamma 2 chain expression was observed clearly in tumor cell cytoplasm. Of the 67 tumors examined, 6 (9%), 31 (46%), 19 (28%), and 11 (17%) showed staining patterns A, B, C, and D, respectively. With progression from staining pattern A to D, the number of immunopositive tumor cells increased significantly (P < 0.0001), and the tumor histology showed significantly more infiltrative growth (P < 0.0001) and poorer differentiation (P = 0.0021). Furthermore, both univariate (P = 0.0019) and multivariate (P = 0.0003; hazard ratio = 3.132) analysis of the patients' survival revealed that the prognosis became significantly poorer with progression from staining pattern A to D. CONCLUSIONS. Increased laminin-5 gamma 2 chain immunoreactivity, which may reflect a high invasive potential of cancer cells, is a factor indicative of a poor prognosis for patients with squamous cell carcinoma of the tongue. Cancer 1999;85:2315-21. (C) 1999 American Cancer Society.
  • T Niki, K Rombouts, P De Bleser, K De Smet, Rogiers, V, D Schuppan, M Yoshida, G Gabbiani, A Geerts
    HEPATOLOGY 29 3 858 - 867 1999年03月 [査読無し][通常論文]
     
    Hepatic stellate cells are the major cellular sources of extracellular matrix in chronic liver diseases leading to fibrosis, We explored the antifibrogenic effect of two histone deacetylase inhibitors, sodium butyrate and trichostatin A (TSA), on this cell type in vitro, Primary hepatic stellate cells as well as culture activated cells were exposed to butyrate (0.01-1 mmol/L) or TSA (1-100 nmol/L); their effect on collagen types I and III and smooth muscle ol-actin was examined by quantitative immunoprecipitation and by Northern analysis. Their antiproliferative effect was examined by H-3-thymidine incorporation and cell counting, Hyperacetylation of histones was demonstrated by acid urea/Triton-X-100 (AUT) polyacrylamide gel electrophoresis. Possible cytotoxic effects were judged on stellate cells by evaluating de novo total protein synthesis, and on hepatocytes by measuring lactate dehydrogenase (LDH) leakage, albumin secretion, and epoxide hydrolase and ethoxycoumarin O-deethylase activity, TSA at 100 nmol/L and butyrate at 1 mmol/L retarded the morphological changes characteristic for activation of primary stellate cells. TSA at 100 nmol/L inhibited synthesis of collagen types I and III and smooth muscle alpha-actin by 62%, 70%, and 88%, Butyrate at 1 mmol/L showed a modest inhibitory effect on collagen type III and smooth muscle alpha-actin, but had no effect on collagen type I. Northern analysis suggested that these inhibitory effects on collagen type III and smooth muscle alpha-actin were transcriptional, while the effect on collagen type I was largely posttranscriptional, At 100 nmol/L, TSA strongly suppressed proliferation of primary hepatic stellate cells. Inhibition of activation of stellate cells was preceded by hyperacetylation of histone H4, When tested on cells at day 14 in culture, butyrate had no inhibitory effects on the synthesis of collagens or smooth muscle alpha-actin. One hundred or 10 nmol/L TSA modestly inhibited the synthesis of collagens type I (-24%,-22%) and III (-34%,-22%), and smooth muscle alpha-actin (-27%,-12%). We conclude that TSA inhibits transdifferentiation of stellate cells into myofibroblasts by interfering with the level of acetylation of histone H4.
  • T Niki, M Pekny, K Hellemans, P De Bleser, K Van den Berg, F Vaeyens, E Quartier, F Schuit, A Geerts
    HEPATOLOGY 29 2 520 - 527 1999年02月 [査読無し][通常論文]
     
    Hepatic stellate cells are considered to be liver-specific pericytes that play a key role in liver fibrosis. Because these cells express desmin and smooth muscle alpha-actin, they were assumed to be of myogenic origin. This hypothesis became doubtful when it was reported that stellate cells also express glial fibrillary acidic protein and neural cell adhesion molecule. In the present study, we show that activated stellate cells express nestin, a class VI intermediate filament protein originally identified as a marker for neural stem cells. Expression of nestin was first studied during spontaneous activation of stellate cells in culture. Immunohistochemistry showed that nestin-positive stellate cells already appeared at day 3, and nearly all the cells became positive for nestin at day 6 and 15. The immunoreaction was present in filaments except in dividing cells. The presence of messenger RNA transcript for nestin was shown by reverse transcription polymerase chain reaction and sequencing of amplified complementary DNA. We then compared the presence of nestin with that of other intermediate filament proteins and smooth muscle alpha-actin, Immunoblotting showed that the relative concentrations of nestin, desmin, and vimentin increased between day 2 and 6 in primary culture. After the initial increase vimentin leveled off, while nestin and desmin showed a tendency to decrease. This pattern was quite different from that of glial fibrillary acidic protein, which kept declining, and smooth muscle alpha-actin, which increased continuously up to day 13 in culture. We then studied the presence of nestin in normal and CCl4-injured rat liver. In normal liver, minimal immunoreaction for nestin was observed within the liver parenchyma. During induction of fibrosis by carbon tetrachloride, nestin-positive stellate cells appeared at 6 weeks, which was late in comparison with the induction of desmin and smooth muscle alpha-actin. We conclude that nestin is induced in stellate cells during transition from the quiescent to the activated phenotype; culture activation is a stronger stimulus than in vivo activation by injection of CCl4. Taken together with reports on expression of glial fibrillary acidic protein and neural cell adhesion molecule by stellate cells, new experimental studies on the embryonic origin of these cells are required.
  • A Geerts, T Niki, K Hellemans, D De Craemer, K Van Den Berg, JM Lazou, G Stange, M Van De Winkel, P De Bleser
    HEPATOLOGY 27 2 590 - 598 1998年02月 [査読無し][通常論文]
     
    In this study, we present a new method to obtain pure, viable, freshly isolated hepatic stellate cells. Stellate cells were purified by cell sorting using their high side scatter (SSC) of incident light. Purity of the cells was established by light and transmission electron microscopy (TEM). Starting from stellate cells that were 50% to 70% enriched by centrifugation in 11% Nycodenz, the cell purity after sorting was found to be 96.6% +/- 2.9%. Viability of the sorted cells was 90.8% +/- 2.2% as measured by the Trypan blue exclusion test and was confirmed by cell culturing. Per hour of sorting, 1.4 +/- 0.4 million stellate cells were obtained. Sorting runs of up to 4 hours were practically feasible, resulting in yields of 5 to 6 million cells per rat liver. Cells attached to plastic substratum within 24 hours. Subsequently, they spread and underwent spontaneous transition into myofibroblast-like cells. The purity of sorted cells was documented by reverse-transcriptase polymerase chain reaction (RT-PCR) experiments using specific primer pairs for messenger RNA (mRNA) species that were only present in parenchymal (preproalbumin), endothelial (endothelial cell nitric oxide synthase [eNOS]), stellate (desmin), or Kupffer cells (77- to 88-kd fucose receptor). Contaminating mRNA species were absent in sorted stellate cells. Next, we examined freshly sorted stellate cells by Western blotting to confirm the presence of relevant cytoskeletal proteins. Cells were positive for vimentin, desmin, and glial fibrillary acidic protein (GFAP), but negative for alpha-smooth muscle actin (alpha-SMA). Sorted and cultured cells were immunophenotyped for the presence of collagen types I, III, and IV, laminin, and the cytoskeletal proteins, alpha-SMA, desmin, vimentin, and GFAP. At 90 hours in culture, cells expressed all the investigated extracellular matrix proteins. Desmin was present in 82% +/- 1%, vimentin in 96% +/- 2.5%, and GFAP in 91% +/- 4.5% of cells. alpha-SMA was present in 91% +/- 2% of cultured cells. We conclude that cell sorting based on SSC of incident light is a convenient method to obtain virtually pure stellate cells that can be used for direct analysis or for culturing. Although the yields obtained with this method are lower than with standard methods, and additional equipment is required, SSC-activated sorting offers the possibility of very pure cells when essential for analyses based on sensitive detection methods such as RT-PCR.
  • PJ DeBleser, T Niki, GX Xu, Rogiers, V, A Geerts
    HEPATOLOGY 26 4 905 - 912 1997年10月 [査読無し][通常論文]
     
    Activins are dimeric proteins, members of the transforming growth factor beta (TGF-beta) gene superfamily, consisting of the beta-subunits of inhibin (beta(A) and beta(B)). Recently, it was shown that activin A (beta(A):beta(A)) inhibits DNA replication and induces apoptosis in rat parenchymal cells in vitro and in vivo. Cryostat sections of normal livers and livers of rats treated with intraperitoneal injections of CCl4 were stained for the different inhibin subunits and desmin, a marker for stellate cells (SC). Staining for inhibin-alpha mas invariably negative, both in normal and fibrotic rat liver. In normal liver, inhibin-beta(A) subunit immunoreactivity was localized in parenchymal cells (PC). Staining for inhibin-beta(B) was weaker but similarly distributed. In fibrotic livers, connective tissue septa were strongly immunoreactive for inhibin-beta A. Desmin-positive stellate cells (SC) accumulated in areas strongly immunoreactive for inhibin-beta A and several cells were positive for both desmin and inhibin-beta(A). staining for inhibin-beta was weaker but similarly distributed. As above data pointed to a possible role for PC and SC, we examined by Northern blot analysis, the expression of inhibin-alpha, -beta(A), and -beta(B) messenger RNA (mRNA) in total RNA extracted from freshly isolated SC and PC of normal and CCl4-treated liver and in cultured SC. Inhibin-beta(A) mRNA was predominantly expressed in PC of normal liver. Expression was lost in PC of CCL4-treated liver. Inhibin-beta(B) mRNA expression was induced in SC of CCl4-treated liver. Inhibin-beta(A) mRNA, and to a lesser extent, inhibin-beta(B) mRNA expression was rapidly induced in cultured SC. The presence of activin A in conditioned media of cultured SC was shown by Western blotting. Apoptotic cells, identified by terminal deoxy-transferase mediated X-dUTP nick end labeling (TUNEL)-staining, were found predominantly in and near the fibrotic septa. In conclusion: 1) while activin A was constitutively expressed in PC of normal liver, its expression was lost in PC of fibrotic liver; 2) expression of activins was induced in activated SC; and 3) apoptotic cells were found predominantly near the septa, in support of the hypothesis that activin A, derived from activated SC in the septa, contributes to the induction of cell death in neighboring PC.
  • GX Xu, T Niki, Virtanen, I, Rogiers, V, P DeBleser, A Geerts
    JOURNAL OF PATHOLOGY 183 1 90 - 98 1997年09月 [査読無し][通常論文]
     
    Fibronectins are multifunctional glycoproteins that are important components of the extracellular matrix in normal and fibrotic liver. Multiple fibronectin isoforms are generated from a single gene by alternative splicing of the primary transcript at the domains EIIIA, EIIIB, and V. The aim of this study was to investigate the fibronectin isoforms expressed by activated hepatic stellate cells, the most important connective tissue-producing cells in injured liver, Hepatocytes and skin fibroblasts were also studied for comparison, Activation of hepatic stellate cells in ville was induced by injecting CCl4 twice weekly for 3 weeks. Activation in vitro was achieved by culturing cells on plastic. The level of activation was evaluated by alpha-smooth muscle actin immunocytochemistry. Steady-state levels of fibronectin isoform messenger RNA were examined by Northern hybridization analysis using specific cDNA probes for the EIIIA, EIIIB, and V domains. The de novo synthesis of fibronectin isoforms was examined by metabolic labelling and immunoprecipitation using domain-specific monoclonal antibodies. Fibronectin transcripts were not detectable in freshly isolated hepatic stellate cells from normal liver. Cultured hepatic stellate cells, as web as skin fibroblasts, expressed EIIIA(+), EIIIB+, and V95(+) transcripts. They were detectable as early as day 3 and increased with time in culture, At 3 days in culture, more than 90 per cent of stellate cells were alpha-smooth muscle actin-positive, In vivo activated hepatic stellate cells expressed EIIIA(+) and V95(+) transcripts; EIIIB+ fibronectin mRNA was absent, Less than 20 per cent of in vice activated stellate cells expressed alpha-smooth muscle actin. Freshly isolated parenchymal cells from normal liver as web as from CCl4-treated liver expressed V95(+) transcripts, but were negative for EIIIA or EIIIB fibronectin mRNA, Immunoprecipitation results were in accordance with Northern hybridization analysis, Hepatic stellate cells in culture synthesized and secreted fibronectin molecules that contained EIIIA, EIIIB, and V fragments, Our results indicate that hepatic stellate cells synthesize and secrete fibronectin isoforms that are distinct from those of parenchymal cells. (C) 1997 by John Wiley & Sons, Ltd.
  • T Niki, D Schuppan, PJ deBleser, R Vrijsen, M PipeleersMarichal, R Beyaert, E Wisse, A Geerts
    HEPATOLOGY 23 6 1673 - 1681 1996年06月 [査読無し][通常論文]
     
    Glucocorticoids have been shown to suppress collagen synthesis and gene expression by fibroblasts. However, little is Known about their effects on fat-storing cells, the major matrix-producing cells in liver fibrosis. In this study we investigated the effect of dexamethasone on the extracellular matrix expression by cultured rat fat-storing cells. Fat-storing cells were isolated from male Wistar rats by collagenase/pronase digestion and purified by density gradient centrifugation. Pat-storing cells in early primary culture (3-day-old, representing a relatively quiescent phenotype) and in subculture (one passage, about a-week-old, representing an activated phenotype) were treated with 10(-6) mol/L dexamethasone for messenger RNA (mRNA) study or with 10(-8) to 10(-6) mol/L dexamethasone for protein study. Expression of collagen type I, III, IV, fibronectin, and laminin was analyzed at the mRNA level by Northern hybridization, and at the protein level by metabolic labeling and immunoprecipitation. Dexamethasone had a variable effect on the expression of collagen alpha 1(I) mRNA level. While a tendency for modest suppression was observed (5%-50%) in primary cells, the difference was not statistically significant, Variable response was observed in subcultured cells. Collagen alpha 1(III) mRNA level showed a tendency for stimulation, Dexamethasone stimulated the expression of collagen alpha 1(IV), fibronectin, and laminin B1 mRNA levels by 1.4-, 2.4-, and 1.6-fold respectively, in primary fat-storing cells. Subcultured cells showed a similar response, but the magnitude of stimulation was more variable than that of primary cells. Unexpectedly, at the protein level dexamethasone had no effect on the expression of these proteins. Our results indicate that glucocorticoids do not possess a net suppressive effect on extracellular matrix synthesis by fat-storing cells. Beneficial effects of glucocorticoids may be attributable to other mechanisms of action, such as their anti-inflammatory effect.
  • T Niki, PJ DeBleser, GX Xu, K VanDenBerg, E Wisse, A Geerts
    HEPATOLOGY 23 6 1538 - 1545 1996年06月 [査読無し][通常論文]
     
    Fat-storing cells are the major producers of extracellular matrix in the liver. A good immunocytochemical marker is, however, still lacking for this cell type. Desmin, frequently used by most investigators, fails to stain many pericentral fat-storing cells in normal rat liver. The aim of the present study is to evaluate glial fibrillary acidic protein (GFAP) as an alternative marker of fat-storing cells. In normal rat liver, immunostaining of GFAP revealed numerous fat-storing cells with characteristic cytoplasmic extensions. Unlike desmin, which was preferentially expressed in periportal fat-storing cells, GFAP-positive fat-storing cells were distributed more evenly in the lobules, In a narrow periportal zone, however, GFAP-positive cells were occasionally absent. Dual GFAP/desmin staining revealed colocalization of these markers, but fat-storing cells positive only for GFAP or desmin were also present, Chronic carbon tetrachloride exposure induced a spatial change in the expression of GFAP and desmin. At 3 weeks, accumulation of GFAP/desmin double-positive cells was observed in developing fibrotic septa. At 8 weeks, the GFAP positivity in the septa persisted but became weak while desmin expression became stronger. In contrast, the expression of GFAP within the lobule was gradually decreased as fibrosis progressed. We conclude that GFAP is expressed by a subpopulation of fat-storing cells, which differs partially from the population that expresses desmin. Because in normal rat liver desmin-negative fat-storing cells can be identified by GFAP staining and vice versa, dual GFAP/desmin staining allows more complete identification of fat-storing cells. In chronically injured liver, GFAP may not be as useful as in normal rat liver. The coexpression of GFAP/desmin in developing septa and the subsequent downregulation of GFAP in an advanced stage of fibrosis may reflect different stages of fat-storing cell activation. Further investigation is required to determine the functional significance of alteration of GFAP expression in fat-storing cells.
  • PJ DEBLESER, P JANNES, SC VANBUULOFFERS, CM HOOGERBRUGGE, CFH VANSCHRAVENDIJK, T NIKI, ROGIERS, V, JL VANDENBRANDE, E WISSE, A GEERTS
    HEPATOLOGY 21 5 1429 - 1437 1995年05月 [査読無し][通常論文]
     
    Transforming growth factor beta (TGF-beta), a potent fibrogenic cytokine, is secreted in latent form. We examined which cell type in both normal and carbon tetrachloride (CCl4)-induced fibrotic rat liver bears surface type IIIGF/mannose 6-phosphate (IGF-II/M6P) receptor, known to facilitate activation of TGF-beta. In addition, the role of the IGF-II/M6P receptor in activation of latent TGF-beta was investigated in a coculture system with sinusoidal endothelial cells. Northern hybridization analysis for IGF-II/M6P receptor messenger RNA (mRNA) was performed on total RNA of different isolated and purified liver cell types. In normal liver, cells expressed little IGF-II/M6P receptor mRNA. In fibrotic liver, we found significant expression only in fat-storing cells. The presence of IGF-II/M6P receptors was established by [I-125]IGF-II binding assays on freshly isolated fat-storing cells from normal and CCl4-exposed rat livers. We found specific binding of [I-125]IGF-II only on CCl4 exposed fat-storing cells. As determined by polyacrylamide gel electrophoresis arter affinity labeling, the specific binding involved 220 kD type II IGF receptors. Scatchard analysis revealed the presence of 3,043 +/- 1,378 IGF-II/M6P high-affinity receptors/fat-storing cell, with a K-d of 387 +/- 165 pmol/L. With a mink lung epithelial cell (Mv(1)Lu) proliferation inhibition assay, inhibition of proliferation (a measure of active TGF-beta function) was determined using conditioned media of activated ht storing cells, cocultures of fat-storing cells, and endothelial cells and pure endothelial cell cultures. We found that conditioned media of cocultures of fat storing cells and endothelial cells inhibits the growth of Mv(1)Lu cells more strongly than conditioned media of homotypic cultures. Addition of neutralizing anti-TGF-beta antibodies neutralizes this effect. The contribution of IGF-II/M6P receptors on the cell membrane of activated fat-storing cells to the activation of latent TGF-beta was demonstrated by incubating the cells with M6P, or antibodies directed to the IGF-II/M6P receptor, both of which diminishes activation of TGF-beta. We conclude that IGF-II/M6P receptors are present on activated fat-storing cells and that the presence of this receptor at the cell surface is necessary, but not sufficient, for activation of latent TGF-beta. Other factors, derived from endothelial cells, are involved.

書籍

  • 病理学・病理検査学
    仁木 利郎 (担当:共編者(共編著者))
    医学書院 2012年03月
  • 標準病理学 第4版
    仁木 利郎 (担当:共編者(共編著者)範囲:細胞傷害の機序とその修復)
    医学書院 2010年08月

講演・口頭発表等

  • 肺癌病理診断の基礎  [通常講演]
    仁木 利郎
    第54回日本肺癌学会総会 2013年11月 公開講演,セミナー,チュートリアル,講習,講義等
  • 現代の医療・医学研究と病理学  [招待講演]
    仁木 利郎
    医の原点 シリーズⅩ 2010年11月 公開講演,セミナー,チュートリアル,講習,講義等
  • 癌と創傷治癒―浸潤先進部の研究から考えたこと  [招待講演]
    仁木 利郎
    第7回日本病理学会カンファランス 2010年08月 口頭発表(招待・特別)
  • 肺癌の病理診断  [通常講演]
    仁木 利郎
    第34回肺癌診断会及び画像診断セミナー 2008年06月 公開講演,セミナー,チュートリアル,講習,講義等

MISC

  • T. Ui, H. Fujii, Y. Hosoya, M. Nagase, M. N. Mieno, M. Mori, T. Zuiki, S. Saito, K. Kurashina, H. Haruta, S. Matsumoto, T. Niki, A. Lefor, Y. Yasuda DISEASES OF THE ESOPHAGUS 28 (2) 180 -187 2015年02月 [査読無し][通常論文]
     
    We retrospectively compared preoperative docetaxel, cisplatin, and fluorouracil (DCF) with cisplatin and fluorouracil (CF) in patients with esophageal cancer. The study included patients with advanced thoracic esophageal carcinoma (excluding T4 tumors) receiving preoperative chemotherapy. In the DCF group, five patients received two courses of treatment every 4 weeks, and 33 patients received three courses every 3 weeks. In the CF group, 38 patients received two courses of treatment every 4 weeks. Patients underwent curative surgery 4-5 weeks after completing chemotherapy. Patient demographic characteristics did not differ between the two study groups. The incidence of a grade 3 or 4 hematologic toxicity was significantly higher in the DCF group (33 patients) than in the CF group (five patients; P < 0.001). Curative resection was accomplished in 79% of patients in the DCF group and 66% in the CF group (P = 0.305). There were no in-hospital deaths. The incidence of perioperative complications did not differ between the groups. A grade 2 or 3 histological response was attained in a significantly higher proportion of patients in the DCF group (63%) than in the CF group (5%; P < 0.001). Progression-free survival and overall survival were significantly higher in the DCF group (P = 0.013, hazard ratio 0.473; P = 0.001, hazard ratio 0.344). In conclusion, a grade 3 or 4 hematologic toxicity was common in the DCF group but was managed by supportive therapy. Histological response rate, progression-free survival, and overall survival were significantly higher in the DCF group compared with the CF group.
  • Daisuke Matsubara, Teppei Morikawa, Akiteru Goto, Jun Nakajima, Masashi Fukayama, Toshiro Niki MODERN PATHOLOGY 22 (6) 776 -785 2009年06月 [査読無し][通常論文]
     
    We report here the presence of subepithelial myofibroblasts in pure bronchioloalveolar carcinoma and a subset of invasive lung adenocarcinoma. The subepithelial myofibroblasts we describe were observed in a peculiar location beneath the cancer cells in the alveolar septa. Immunohistochemically, they were positive for alpha-smooth muscle actin and calponin, but negative for desmin and h-caldesmon. To gain insight into their biological significance, we examined 116 surgically resected lung adenocarcinomas. The resected tumors included 13 bronchioloalveolar carcinomas, 20 mixed type adenocarcinomas with bronchioloalveolar carcinoma components, 57 papillary adenocarcinomas, 22 solid adenocarcinomas with mucin, and 4 acinar adenocarcinomas. All specimens were immunostained for alpha-smooth muscle actin to visualize the myofibroblasts. In all of the pure bronchioloalveolar carcinomas observed, the subepithelial myofibroblasts were completely preserved adjacent to the cancer cells. In mixed adenocarcinomas with bronchioloalveolar carcinoma components, subepithelial myofibroblasts were present in the bronchioloalveolar carcinoma components, but scanty in the invasive areas, where stromal myofibroblasts emerged between the cancer cell nests. Subepithelial myofibroblasts were retained, however, in the invasive areas of a subset of invasive adenocarcinomas. Survival analysis showed that the retention of subepithelial myofibroblasts in these invasive tumors was associated with low rates of lymphatic and vascular invasion, a low rate of lymph node involvement, and an excellent patient survival. These results suggest that subepithelial myofibroblasts increase in bronchioloalveolar carcinomas, but are gradually replaced by typical stromal myofibroblasts during progression into invasive cancer. A subset of invasive adenocarcinomas retains subepithelial myofibroblasts. Analysis of subepithelial myofibroblasts may be helpful in identifying a subset of lung adenocarcinoma with excellent prognosis. Modern Pathology (2009) 22, 776-785; doi: 10.1038/modpathol.2009.27; published online 27 March 2009
  • Hidenori Haruta, Yoshinori Hosoya, Kazuya Sakuma, Hiroyuki Shibusawa, Kiichi Satoh, Hironori Yamamoto, Akira Tanaka, Toshiro Niki, Kentaro Sugano, Yoshikazu Yasuda JOURNAL OF DIGESTIVE DISEASES 9 (4) 213 -218 2008年11月 [査読無し][通常論文]
     
    BACKGROUND: The endoscopic resection of early gastric cancers (EGC) is a standard technique in Japan and is increasingly used throughout the world. Further experience in the treatment of EGC and a clearer delineation of the factors related to lymph-node metastasis would permit a more accurate assessment of endoscopic resection. METHODS: The study group comprised 1389 patients with EGC who underwent gastrectomy with lymph-node dissection. We evaluated the relations of lymph-node metastasis to clinicopathological factors. RESULTS: Of the 718 patients with intramucosal carcinomas, 14 (1.9%) had lymph-node metastasis. All cases of lymph-node metastasis were associated with ulceration. No lymph-node metastasis was found in patients with intramucosal carcinomas without ulceration, irrespective of tumor size and histological type. Lymph-node metastasis was present in 14 (4.7%) of the 296 patients who had cancer with a submucosal invasion depth of less than 500 mu m (sm 1). Significantly increased rates of lymph-node metastasis were associated with undifferentiated types, ulcerated lesions and lymphatic invasion. No lymph-node metastasis was found in patients with differentiated sm 1 carcinomas 30 mm or less in diameter without ulceration. Lymph-node metastasis occurred in 29% of the patients who had cancer with a submucosal invasion depth of 500 mu m or more (sm 2). CONCLUSION: This large series of patients with EGC provides further evidence supporting the expansion of indications for endoscopic treatment, as well as warns against potential risks.
  • Li CP, Goto A, Watanabe A, Murata K, Ota S, Niki T, Aburatani H, Fukayama M Pathol Res Pract. 2004 295-304. 2008年 [査読無し][通常論文]
  • Inamura K, Takeuchi K, Togashi Y, Nomura K, Ninomiya H, Okui M, Satoh Y, Okumura S, Nakagawa K, Soda M, Lim CY, Niki T, Mano H, Ishikawa J Thorac Oncol. 3 13-7. 2008年 [査読無し][通常論文]
  • Yu Nakamura, Daisuke Matsubara, Akiteru Goto, Satoshi Ota, Oguni Sachiko, Shumpei Ishikawa, Hiroyuki Aburatani, Keiji Miyazawa, Masashi Fukayama, Toshiro Niki CANCER SCIENCE 99 (1) 14 -22 2008年01月 [査読無し][通常論文]
     
    In this study we explored the mechanisms of constitutive activation of c-Met in lung adenocarcinoma cell lines. First, we examined levels of c-Met and phospho-c-Met (Y1234/Y1235) in a panel of lung adenocarcinoma cell lines by Western blot analysis. c-Met expression was found in 12 of 14 cell lines and an overall correlation between the expressions of c-Met and phospho-c-Met was noted. c-Met was constitutively activated particularly at high levels in five cell lines (PC3, LC-2/ad, L27, H1648, and H2009). c-Met amplification was identified in L27 and H1648 by single nucleotide polymorphism array analysis, but no mutations were identified in the Sema domain or in any part of the cytoplasmic domain of c-Met. Experiments with neutralizing anti-hepatocyte growth factor (HGF) antibody, scatter assay using Madin-Darby canine kidney cells, and Western blotting on conditioned media of the cell lines revealed that the constitutive phosphorylation of c-Met was largely ligand-independent. The inhibition of cell-matrix adhesion induced the dephosphorylation of c-Met in the five cell lines tested. This was accompanied by downregulation of c-Met in three of the five cell lines. In contrast, the inhibition of cell-cell adhesion by neutralizing E-cadherin antibody had a minimal effect on the expression and phosphorylation of c-Met. These results reveal three features of the constitutive activation of c-Met in our panel of lung adenocarcinoma cell lines: (i) it correlates with c-Met overexpression, either with or without gene amplification; (ii) it is largely ligand-independent; and (iii) it depends on cell-matrix adhesion.
  • Kengo Murata, Satoshi Ota, Toshiro Niki, Akiteru Goto, Chih-Ping Li, Urbiztondo Maria Rhea Ruriko, Shumpei Ishikawa, Hiroyuki Aburatani, Takayuki Kuriyama, Masashi Fukayama EXPERIMENTAL AND MOLECULAR PATHOLOGY 83 (3) 367 -376 2007年12月 [査読無し][通常論文]
     
    Idiopathic pulmonary fibrosis (IPF) is the most common lung disease predisposing lung cancer. To clarify the early phase of epithelial abnormalities in IPF, we used an in vitro squamous metaplasia model, transforming growth factor beta 1 (TGF beta 1)-treated airway epithelial cells (BEAS-2B). The model repeated the expression of squamous epithelial character, such as involucrin, and keratin 6 and 14. DNA microarray analysis disclosed a unique expression signature in TGF beta 1-treated airway epithelial cells, 20 specifically up-regulated genes including p63,jagged 1 (jag1) and the genes of structure proteins. Western blotting and RT-PCR analysis revealed that Delta Np63 alpha was the dominant isoform of p63 in our experimental model. Immunohistochemical analysis demonstrated the expression of p63 and jag1 in lung tissues of IPF. Inhibition of p63 with siRNA caused the down-regulation of jag1 expression, but not of involucrin, or keratin 6 and 14. Interestingly, the up-regulation of p63 was totally suppressed by N-acetyl-L-cysteine (NAC), but not by dexamethasone or pirfenidone. Thus, the p63-jag1 pathway may be up-regulated at an early phase of epithelial abnormalities in IPF, which can be overcome by NAC even in the TGF beta 1-rich milieu. (c) 2007 Elsevier Inc. All rights reserved.
  • Manabu Soda, Young Lim Choi, Munehiro Enomoto, Shuji Takada, Yoshihiro Yamashita, Shunpei Ishikawa, Shin-ichiro Fujiwara, Hideki Watanabe, Kentaro Kurashina, Hisashi Hatanaka, Masashi Bando, Shoji Ohno, Yuichi Ishikawa, Hiroyuki Aburatani, Toshiro Niki, Yasunori Sohara, Yukihiko Sugiyama, Hiroyuki Mano NATURE 448 (7153) 561 -U3 2007年08月 [査読無し][通常論文]
     
    Improvement in the clinical outcome of lung cancer is likely to be achieved by identification of the molecular events that underlie its pathogenesis. Here we show that a small inversion within chromosome 2p results in the formation of a fusion gene comprising portions of the echinoderm microtubule-associated protein-like 4 (EML4) gene and the anaplastic lymphoma kinase (ALK) gene in non-small-cell lung cancer (NSCLC) cells. Mouse 3T3 fibroblasts forced to express this human fusion tyrosine kinase generated transformed foci in culture and subcutaneous tumours in nude mice. The EML4-ALK fusion transcript was detected in 6.7% (5 out of 75) of NSCLC patients examined; these individuals were distinct from those harbouring mutations in the epidermal growth factor receptor gene. Our data demonstrate that a subset of NSCLC patients may express a transforming fusion kinase that is a promising candidate for a therapeutic target as well as for a diagnostic molecular marker in NSCLC.
  • Yu Nakamura, Toshiro Niki, Akiteru Goto, Teppei Morikawa, Keiji Miyazawa, Jun Nakajima, Masashi Fukayama CANCER SCIENCE 98 (7) 1006 -1013 2007年07月 [査読無し][通常論文]
     
    c-Met is often overexpressed in non-small cell lung cancer, but it remains unsolved whether its overexpression leads to its activation. We used an antibody specific to phospho-c-Met (Tyr1235) to investigate c-Met activation immunohistochemically in 130 surgically resected lung adenocarcinomas. The expression of c-Met and hepatocyte growth factor (HGF) was also investigated. Phospho-c-Met was positive in 21.5% (28/130) of cases. c-Met was positive in 74.6% of cases (97/130) and was expressed at high levels in 36.1% of cases (47/130). HGF was expressed at high levels in 31.5% of cases (41/130). Phospho-c-Met was correlated with high levels of HGF (P = 0.0010) and high levels c-Met expression (P = 0.0303), but it was also found to be positive in 12 cases with little to no HGF expression. Phospho-c-Met expression was significantly associated with tumor differentiation (P = 0.0023) and papillary histology (P = 0.0011), but not with pathological stage, lymph node metastasis or survival. High levels of c-Met and HGF were also associated with papillary histology (P = 0.0056 and P = 0.0396, respectively), but not with tumor differentiation. Phospho-c-Met was correlated with phospho-Akt (P = 0.0381), but not with phospho-Erk or phospho-Stat3. Phospho-Akt expression was marginally correlated with the expression of phospho-epidermal growth factor receptor (EGFR) (P = 0.0533) and, importantly, it was strongly correlated with the expression of either phospho-c-Met or phospho-EGFR (P = 0.0013). The data suggest that in lung adenocarcinoma tissue, c-Met activation may take place either ligand-dependently or ligand-independently via c-Met overexpression. c-Met activation may play special roles in the papillary subtype and in well differentiated lung adenocarcinomas.
  • Tao Wang, Toshiro Niki, Akiteru Goto, Satoshi Ota, Teppei Morikawa, Yu Nakamura, Etsuko Ohara, Shumpei Ishikawa, Hiroyuki Aburatani, Jun Nakajima, Masashi Fukayama CANCER SCIENCE 98 (4) 506 -511 2007年04月 [査読無し][通常論文]
     
    Tumor hypoxia is associated with a malignant phenotype of cancer cells and poor patient prognosis. To investigate the role of hypoxia in tumor progression, we studied the effects of hypoxia in the A549 lung adenocarcinoma cell line. First, we showed that hypoxic treatment decreased cell-cell adhesion and induced a scattering of cancer cells. Concomitant with these morphological changes, the motility of cancer cells was increased, as demonstrated by the Boyden chamber assay. Then, we used oligonucleotide array analyses to identify the genes causally related to the hypoxia-induced motile phenotype. The results showed that the expression of approximately 100 genes was induced more than 5-fold by hypoxia. These included (among others) epidermal growth factor receptor (EGFR), as well as other well-known hypoxia-induced genes, such as vascular endothelial growth factor. Immunohistochemical analyses of primary lung adenocarcinomas confirmed the induction of EGFR in tumor cells in the vicinity of necrotic areas, a histological indicator of tumor hypoxia. Remarkably, the EGFR inhibitor AG1478 (10 mu M) completely blocked the increased cell motility induced by hypoxia. Thus, the present study demonstrates the importance of the EGFR pathway in the increased motility of cancer cells that occurs in a hypoxic tumor environment.
  • T. Narahashi, T. Niki, T. Wang, A. Goto, D. Matsubara, N. Funata, M. Fukayama HISTOPATHOLOGY 49 (4) 349 -357 2006年10月 [査読無し][通常論文]
     
    Aims: To determine the significance of p63 protein expression in the development and progression of lung adenocarcinoma. Methods and results: The expression of p63 was immunohistochemically investigated in 92 cases of lung adenocarcinoma with a maximum diameter of 30 mm or less. p63 expression was observed not only in the nuclei (46/92 cases, 50%), but also in the cytoplasm of neoplastic cells (47/92, 51%). Nuclear localization of p63 was correlated with nuclear accumulation of p53 (P = 0.0120), whereas the presence of nuclear p63 had no apparent effect on patient survival. Cytoplasmic localization of p63 was found to be correlated with shorter survival periods by univariate and multivariate analyses (P = 0.0486 and P = 0.0488, respectively) and the relation was independent of clinicopathological factors. Cytoplasmic localization of p63 was further confirmed by immunoblots of the cytoplasmic fraction of HLC-1, a lung adenocarcinoma cell line which predominately expressed Delta Np63 alpha transcript relative to TAp63 transcript by quantitative reverse transcriptase-polymerase chain reaction. Conclusions: Cytoplasmic expression of p63 is an adverse prognostic factor in patients with adenocarcinoma of the lung.
  • D Matsubara, T Niki, S Ishikawa, A Goto, E Ohara, T Yokomizo, CW Heizmann, H Aburatani, S Moriyama, H Moriyama, Y Nishimura, N Funata, M Fukayama CANCER SCIENCE 96 (12) 844 -857 2005年12月 [査読無し][通常論文]
     
    Previous studies suggest that some S100 proteins are involved in the progression of certain types of cancer. However, no comprehensive data is currently available on the expression of S100 family genes in lung adenocarcinomas. Oligonucleotide array, quantitative reverse transcription-polymerase chain reaction and western blot analyses of lung adenocarcinoma cell lines and bronchiolar epithelial cells (SAEC and NHBE) revealed that S100A2 and S100A4 were the most strikingly downregulated and upregulated members of the S100 family, respectively. Immunohistochemical analyses of 94 primary lung adenocarcinomas showed that positive S100A2 expression (33/94, 35.1%) was significantly associated with lymphatic invasion (P = 0.0233) and positive S100A4 expression (19/94, 20.2%) with vascular invasion (P=0.0454). Interestingly, a strong inverse relationship was found between S100A4 and p53 expression (P = 0.0008). Survival analyses showed that S100A4 positivity was associated with poor patient prognosis (P = 0.042). S100A2 positivity was not associated with patient survival when the whole patient group was analyzed; however, S100A2 positivity was a favorable prognostic indicator in patients with p53-negative tumors (P = 0.0448). Finally, we used oligonucleotide array analyses and identified potential S100A2 and S100A4 target genes involved in cancer progression: S100A2 induced RUNX3 and REPRIMO; S100A4 induced EZRIN, RUNX1 and WISP1; S100A2 repressed EGFR, NFKB2 and RELA2; and S100A4 repressed ANXA10 and IL1RN. Thus, the present study demonstrates involvement of S100A2 and S100A4 in the progression of lung adenocarcinomas and an inverse association between S100A4 and p53 expression, and provides a list of targets regulated by S100A2 and S100A4.
  • N Yamauchi, A Watanabe, M Hishinuma, K Ohashi, Y Midorikawa, Y Morishita, T Niki, J Shibahara, M Mori, M Makuuchi, Y Hippo, T Kodama, H Iwanari, H Aburatani, M Fukayama MODERN PATHOLOGY 18 (12) 1591 -1598 2005年12月 [査読無し][通常論文]
     
    Expression profiling of hepatocellular carcinoma has demonstrated that glypican 3 (GPC3), a heparan sulfate proteoglycan anchored to the membrane, is expressed at a markedly elevated level in hepatocellular carcinoma. In this paper, two monoclonal antibodies against GPC3, GPC3-C02 and A1836A, were confirmed to specifically recognize GPC3molecule in cells from hepatocellular carcinoma and hepatoblastoma cell lines by immunoblotting, and both were confirmed to recognize different epitopes of the GPC3 molecule by epitope mapping. Then, we evaluated the feasibility of GPC3-immunohistochemistry in the pathological diagnosis of benign and malignant hepatocellular lesions by applying these monoclonal antibodies to formalin-fixed and paraffin-embedded specimens. The immunoreactivity turned out to be identical in the two monoclonal antibodies and was thus confirmed to represent the actual expression of the GPC3 molecule. The expression was observed in the fetal liver, but not in normal adult liver, liver cirrhosis or hepatitis except for a tiny focus of a regenerative nodule of fulminant hepatitis. Diffusely positive staining of GPC3 was observed in malignant hepatocytes in hepatoblastomas and in hepatocellular carcinomas (47/56, 84%). GPC3 expression was independent of the differentiation and size of the hepatocellular carcinoma. On the other hand, there was only weak and focal staining in low-grade (2/8) and high-grade dysplastic nodules (6/8). GPC3 immunoreactivity was detected in only one of 23 metastatic lesions of colorectal carcinoma, and its expression was entirely absent in the liver cell adenoma (0/7), carcinoid tumor (0/1), and cholangiocellular carcinoma (0/16). When compared with immunohistochemistry of hepatocyte antigen and alpha-fetoprotein, GPC3-immunohistochemistry was siginificantly much more specific and sensitive for hepatocellular carcinomas. Thus, GPC3 was confirmed to be one of the oncofetal proteins now attracting attention for their promise both as markers of hepatocellular carcinoma in routine histological examination and as targets in monoclonal antibody-based hepatocellular carcinoma therapy.
  • A Goto, T Niki, CP Li, D Matsubara, Y Murakami, N Funata, M Fukayama CANCER SCIENCE 96 (8) 480 -486 2005年08月 [査読無し][通常論文]
     
    The TSLC1 (tumor suppressor in lung cancer 1) gene is a tumor suppressor recently identified through functional complementation in a lung adenocarcinoma cell line A549. In this study we immunohistochemically examined the loss of TSLC1 expression in 93 cases of surgically resected lung adenocarcinoma, and investigated its correlation with clinicopathological parameters, including histological subtypes of tumors. The prognostic significance of loss of TSLC1 expression was analyzed by univariate and multivariate analyses, in parallel with other prognostic markers such as p53, p27, and Ki-67. In non-cancerous lung tissue, TSLC1 was weakly positive in bronchial and bronchiolar epithelial cells, type II pneumocytes and bronchial glands. Overall, TSLC1 was negative in 60 of 93 lung adenocarcinomas. TSLC1 was mainly localized in the cytoplasm of the cells, but cell membrane staining was also observed, especially at sites of cell-cell adhesion. TSLC1-negative tumors were more frequently observed in male cases (41/54 cases, 70.0%) than in female cases (19/39 cases, 48.7%) (P<0.01). Notably, TSLC1 expression was preserved in a non-invasive, bronchiolo-alveolar histological pattern of tumor cells (P<0.0001). Survival analyses showed that loss of TSLC1 expression was associated with lower patient survival in univariate and multivariate analyses (P<0.05 and P=0.059, respectively). Subset analyses further showed that the prognostic impact of loss of TSLC1 was significant for male patients (P=0.0089), but not for female patients. We conclude that TSLC1 is expressed in a subset of lung adenocarcinomas, especially in those with bronchiolo-alveolar spread pattern. Loss of TSLC1 is associated with lower patient survival, supporting its role as a tumor suppressor.
  • Soft and hard keratin expression in Epstein-Barr-virus-associated gastric carcinoma.
    Uozaki H, Chong JM, Fujimoto E, Itoh M, Saito M, Sakuma K, Sudo M, Ushiku T, Niki T, Nagai H, Takada K, Fukayama M Anticancer Res. 25 3183-3190. 2005年 [査読無し][通常論文]
  • A Goto, J Nakajima, K Hara, T Niki, M Fukayama VIRCHOWS ARCHIV 446 (1) 73 -77 2005年01月 [査読無し][通常論文]
     
    A 46-year-old man presented with a lung tumor 17 years after a subtotal colectomy and 13 years after a partial duodenectomy for familial adenomatous polyposis (FAP). There had been no malignant transformation in the specimens from his colectomy and duodenectomy, and a current gastrointestinal investigation revealed no evidence of malignancy. Pathological analysis of the lung tumor demonstrated adenocarcinoma with clear cells and a papillary structure, accompanied by tiny tumorous nodules in the background lung parenchyma. Many of the nodules were multifocal adenocarcinoma; however, some of the nodules demonstrated atypical adenomatous hyperplasia (AAH). This is the first case report of a lung adenocarcinoma accompanied by AAH in a FAP patient. Immunohistochemical and loss of heterozygosity studies revealed unique features of the lesions reflecting a disruption of the adenomatous poliposis coli - beta- catenin pathway.
  • Fukumoto S, Yamauchi N, Moriguchi H, Hippo Y, Watanabe A, Shibahara J, Taniguchi H, Ishikawa S, Ito H, Yamamoto S, Iwanari H, Hironaka M, Ishikawa Y, Niki T, Sohara Y, Kodama T, Nishimura M, Fukayama M, Dosaka-Akita H, Aburatani H Clin Cancer Res 11 1776-1785. 2005年 [査読無し][通常論文]
  • Goto A, Niki T, Moriyama S, Funata N, Moriyama H, Nishimura Y, Tsuchida R, Kato JY, Fukayama M Pathol Int. 54 (9) 675 -681 2004年 [査読無し][通常論文]
  • Goto A, Niki T, Terado Y, Fukushima J, Fukayama M Histopathology. 45 384-392. 2004年 [査読無し][通常論文]
  • Shibahara J, Goto A, Niki T, Tanaka M, Nakajima J, Fukayama M Am J Surg Pathol. 28 825-829. 2004年 [査読無し][通常論文]
  • Kitagawa H, Goto A, Niki T, Hironaka M, Nakajima J, Fukayama M Pathol Int. 53 (12) 823 -827 2003年 [査読無し][通常論文]
  • Kitagawa H, Goto A, Minami M, Nakajima J, Niki T, Fukayama M Jpn J Clin Oncol. 33 (7) 360 -363 2003年 [査読無し][通常論文]
  • Terasaki H, Niki T, Matsuno Y, Yamada T, Maeshima A, Asamura H, Hayabuchi N, Hirohashi S Am J Surg Pathol 27 937-951. 2003年 [査読無し][通常論文]
  • H Takei, H Asamura, A Maeshima, K Suzuki, H Kondo, T Niki, T Yamada, R Tsuchiya, Y Matsuno JOURNAL OF THORACIC AND CARDIOVASCULAR SURGERY 124 (2) 285 -292 2002年08月 [査読無し][通常論文]
     
    Objective: Large cell neuroendocrine carcinoma of the lung is a newly recognized clinicopathologic entity. The clinical characteristics and optimal treatment of patients with large cell carcinomas are not yet established. The aim of this study was to define the clinicopathologic characteristics of large cell neuroendocrine carcinoma. Methods: The histologic characteristics of the patients receiving an initial diagnosis of poorly differentiated non-small cell lung carcinoma (n = 484), small cell carcinoma (n = 55), carcinoid (n = 31), and large cell neuroendocrine carcinoma (n = 12) were retrospectively reviewed according to World Health Organization criteria. Immunohistochemistry was performed to confirm the neuroendocrine phenotype. The outcomes and other clinical characteristics of those patients with large cell neuroendocrine carcinoma were retrospectively analyzed and compared with those of patients with poorly differentiated carcinoma of other histologic types. Results: A total of 87 patients were given a diagnosis of large cell neuroendocrine carcinoma after the histologic review. These patients comprised 3.1% of all patients undergoing resection for primary lung cancer during the same period. The overall 5-year survival was 57%. The 5-year survivals of patients with stage I, II, III, and IV disease were 67%. 75%, 45%, and 0%, respectively. There was no statistically significant difference between the overall survival of patients with large cell neuroendocrine carcinoma and those with other non-small cell lung cancers. There was a significant difference between the survival of patients with stage I large cell neuroendocrine carcinoma and that of patients with the same stage of other non-small cell lung carcinomas. The site of the first documented recurrence was locoregional in 12 patients (34%), distant metastases in 20 patients (57%), and both simultaneously in 3 patients. Locoregional lymph node recurrences were observed frequently. More than 80%. of recurrences were found within 1 year after the operation. Conclusion: in terms of prognosis, large cell neuroendocrine carcinoma is distinctly different from other non-small cell lung cancers. The prognosis of large cell neuroendocrine carcinoma was poor, even for early stage disease; the prognosis of the stage I disease of large cell neuroendocrine carcinoma was poorer than that of the same stage of other non-small cell lung cancers. Because of its aggressive clinical behavior and poor prognosis, large cell neuroendocrine carcinoma should be recognized as one of the poorest prognostic subgroups among primary lung cancers, and therefore novel therapeutic approaches should be established.
  • K Rombouts, T Niki, P Greenwel, A Vandermonde, A Wielant, K Hellemans, P De Bleser, M Yoshida, D Schuppan, M Rojkind, A Geerts EXPERIMENTAL CELL RESEARCH 278 (2) 184 -197 2002年08月 [査読無し][通常論文]
     
    Excessive production of collagens by a-smooth muscle actin (alpha-SMA)-positive myofibroblasts leads to fibrotic skin diseases, such as hypertrophic scarring. This process is characterized by an imbalance between extracellular matrix (ECM) synthesis and degradation, while transforming growth factor beta (TGF-beta(1)), known to be a key mediator of fibrogenesis, is up-regulated. In this study we have investigated the possible antifibrogenic effect of Trichostatin A (TSA), a histone deacetylase inhibitor, on rat skin fibroblasts in culture. mRNA steady-state levels and de novo protein synthesis of procollagen types I and III and alpha-SMA were inhibited when skin fibroblasts were treated with 100 nM TSA with or without TGF-beta(1). While the transcription rate of the procollagen alpha1(I) gene was increased following TSA or TGF-beta(1) treatment, TSA abrogated the stimulatory effect of TGF-beta(1) on procollagen alpha1(I) transcription when both compounds were added simultaneously. The reduction of procollagen alpha1(I) and alpha1(III) mRNA steady-state levels by TSA did not require de novo protein synthesis; while the effect of TSA on alpha-SMA mRNA steady-state levels was cycloheximide-sensitive. Interestingly, TSA affected TGF-beta(1) and its downstream mediators, i.e., the Smad family proteins. TSA strongly induced in a biphasic way the expression of 5'TG3' interacting factor (TGIF), a known endogenous corepressor molecule of the TGF-beta(1) signaling pathway. Addition of exogenous TGF-beta(1) did not interfere with the effect of TSA on the TGIF mRNA level. Our study shows that inhibition of histone deacetylases by TSA reduces expression of fibrosis-related genes in skin fibroblasts and this coincides by alterations in the TGF-beta(1) signaling pathway. (C) 2002 Elsevier Science (USA).
  • Frequent Co-Localization of Cox-2 and Laminin-s gamma2 Chain at the Invasive Front of Early-Stage Lung Adenocarcinomas.
    Am J Pathol 160,1129-1141 2002年 [査読無し][通常論文]
  • Clinicopathological significance of epigenetic inactivation of RASSFIA at 3p2l.3 in stage I lung adenocarcinoma.
    Crin Cancer Res. 8, 2362-2368 2002年 [査読無し][通常論文]
  • Proc Natl Acad Sci U S A. 99, 12269-12274 2002年 [査読無し][通常論文]
  • c-MET expression in myofibroblasts: role in autocrine activation and prognostic significance in lung adenocarcinoma.
    Am J Pathol 158, 1451-1463 2001年 [査読無し][通常論文]
  • Expression of vascular endothelial growth factors A, B, C, and D and their relationships to lymph node status in lung adenocarcinoma.
    Clin Cancer Res 6, 2431 -2439 2000年 [査読無し][通常論文]
  • Biochem Biophys Res Commun 275, 440-445 2000年 [査読無し][通常論文]
  • Altered expression of the ERM proteins in lung adenocarcinoma.
    Lab Invest 80, 1643-1650 2000年 [査読無し][通常論文]
  • Clinicopathologic significance of laminin-5 gamma2 chain expression in squamous cell carcinoma of the tongue. Immunohistochemical analysis of 67 lesions.
    Cancer 85, 2315-2321 1999年 [査読無し][通常論文]
  • J Hepatol 26, 886-893 1997年 [査読無し][通常論文]
  • Hepatology 26, 905-912 1997年 [査読無し][通常論文]

共同研究・競争的資金等の研究課題

  • Microenvironment
    Basic Science Research Program
    研究期間 : 1997年 -2020年
  • Epithelial-Stromal Interaction
    Basic Science Research Program
    研究期間 : 1997年 -2020年
  • Cancer and Wound Healing
    Basic Science Research Program
    研究期間 : 1997年 -2020年
  • 肺腺癌の上皮間葉転換におけるエピジェネティックス
    日本学術振興会:科学研究費補助金基盤B一般
    研究期間 : 2014年04月 -2018年03月 
    代表者 : 仁木利郎
  • 肺腺癌の系統的転移モデルの作製
    日本学術振興会:科学研究費補助金挑戦的萌芽研究
    研究期間 : 2013年04月 -2015年03月 
    代表者 : 仁木利郎
  • 微小乳頭癌における分子標的の同定と治療への応用
    日本学術振興会:科学研究費補助金挑戦的萌芽研究
    研究期間 : 2011年04月 -2013年03月 
    代表者 : 仁木利郎
  • 上皮間葉転換形質を示す肺癌における分子標的の同定と
    日本学術振興会:科学研究費補助金基盤B一般
    研究期間 : 2008年04月 -2012年03月 
    代表者 : 仁木 利郎
  • 肺腺癌の浸潤部で誘導される創傷治癒関連分子の同定とその制御機構の破綻について:遺伝子異常との関連性に焦点をあてた検討
    日本学術振興会:日本学術振興会基盤研究C(2)
    研究期間 : 2005年04月 -2007年03月 
    代表者 : 仁木 利郎
  • 肺腺癌の浸潤先端部に発現する炎症・組織修復に関連
    日本学術振興会:日本学術振興会基盤研究C(2)
    研究期間 : 2002年04月 -2004年03月 
    代表者 : 仁木 利郎
  • ヒト肺腺がんにおけるがん間質相互作用について
    日本学術振興会:日本学術振興会基盤研究C(2)
    研究期間 : 1998年04月 -2001年03月 
    代表者 : 仁木 利郎

委員歴

  • 2013年04月 - 現在   日本肺癌学会   国際委員会委員
  • 2013年04月 - 現在   日本病理学会   研究推進委員会委員
  • 2009年02月 - 現在   日本肺癌学会   病理診断委員会委員
  • 2009年04月 - 2014年03月   日本病理学会   病理診断講習会委員会
  • 2008年 - 2009年   日本病理学会   第26回、27回病理専門医試験委員会委員長
  • 2006年 - 2006年   日本病理学会   第24回病理専門医試験実施委員会委員長

担当経験のある科目

  • 病理学実習自治医科大学, 東京大学
  • 病理学総論自治医科大学, 東京大学


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