研究者総覧

武藤 弘行 (ムトウ ヒロユキ)

  • 情報センター 教授
Last Updated :2021/11/23

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

  • 胃癌   

研究分野

  • ライフサイエンス / 消化器内科学

研究活動情報

論文

  • Miho Sashikawa Kimura, Hiroyuki Mutoh, Kentaro Sugano
    JOURNAL OF GASTROENTEROLOGY 46 11 1292 - 1299 2011年11月 [査読有り][通常論文]
     
    Background SOX9 is a marker for stem cells in the intestine and overexpression of SOX9 is found in some types of cancer. However, the expression of SOX9 in normal stomach, precancerous intestinal metaplasia, and gastric carcinoma has not yet been clarified. This study aimed to investigate SOX9 expression in the corpus and pyloric regions of the normal human stomach, premalignant intestinal metaplasia, and gastric carcinoma by using immunohistochemistry. Methods We evaluated SOX9 expression in 46 clinical samples (early gastric well-differentiated adenocarcinoma including surrounding intestinal metaplasia) resected under esophagogastroduodenoscopy. Results A small amount of SOX9 was expressed in the neck/isthmus of the corpus region and SOX9 expression was predominantly restricted to the neck/isthmus of the pyloric region in normal human stomach. In the intestinal metaplastic mucosa, SOX9- and PCNA-positive cells were located at the base of the intestinal metaplastic mucosa. Almost all of the gastric carcinoma cells expressed SOX9. Conclusion SOX9 is expressed in intestinal metaplasia and gastric carcinoma in humans.
  • Hirotsugu Sakamoto, Hiroyuki Mutoh, Hiroko Hayakawa, Miho Sashikawa, Kentaro Sugano
    JOURNAL OF GASTROENTEROLOGY 46 5 620 - 628 2011年05月 [査読有り][通常論文]
     
    Gene expression in the early stage of the transition to intestinal metaplasia in human gastric mucosa has not been determined. In this study, we investigated the temporal relationship between cell lineage changes and intestine-specific gene expression in the process leading to intestinal metaplasia, using Cdx2-transgenic mice. Cellular phenotypes were analyzed by immunohistochemistry and were compared with the gene expression profiles of cell lineage markers by real-time polymerase chain reaction. Up to postnatal day (PD) 20, the gastric mucosae of Cdx2-transgenic mice were histologically similar to those of their normal littermates. However, at approximately PD 20, we observed the sporadic appearance of glands in which all the epithelial cells expressed Cdx2 (Cdx2-diffuse positive glands). In the Cdx2-diffuse positive glands, parietal cells had disappeared, the proliferating zone had moved from the isthmus to the base, and absorptive cells and goblet cells were recognized. In contrast, the surrounding mucosa retained the phenotype of the gastric gland in which only some of the epithelial cells expressed Cdx2. During PDs 30 and 40, the entire fundic mucosa changed to transdifferentiated mucosa that was a composite of intestinal metaplasia and spasmolytic polypeptide-expressing metaplasia. An increase in the expression of intestine-specific genes, with a reciprocal decrease in gastric-specific gene expression, began much earlier than the emergence of Cdx2-diffuse positive glands. A dramatic increase in intestine-specific gene expression precedes the morphological appearance of intestinal metaplasia and spasmolytic polypeptide-expressing metaplasia.
  • Hiroyuki Mutoh, Miho Sashikawa, Kentaro Sugano
    DIFFERENTIATION 81 2 92 - 98 2011年02月 [査読有り][通常論文]
     
    Sox2 is closely related to the gastric phenotype. Sox2 plays a pivotal role in gastric epithelial differentiation in the adult. Sox2 expression is reduced in Helicobacter pylori-associated intestinal metaplastic change of the gastric epithelium. The gastric mucosa is replaced by intestinal metaplastic mucosa in the stomach of caudal type homeobox 2 (Cdx2)-transgenic mice. The aim of this study was to use Cdx2-transgenic mice to investigate: (i) Sox2 expression in the intestinal metaplastic mucosa of the Cdx2-transgenic mouse stomach; and (ii) the relationship between Sox2 and Cdx2. Quantitative real-time PCR was performed to determine Sox2, Cdx2, Muc5Ac, and alkaline phosphatase mRNA expression levels and single- or double-label immunohistochemistry was used to evaluate the localization of Sox2, Cdx2, gastric mucin and alkaline phosphatase activity. We determined that Sox2 mRNA in the intestinal metaplastic mucosa of the Cdx2-transgenic mouse stomach was expressed 3.5-fold compared to the normal mouse stomach. Immunohistochemical analysis showed that the same cells in the intestinal metaplastic mucosa expressed both Cdx2 and Sox2. Gastric mucin was not expressed while alkaline phosphatase activity was recognized in the intestinal metaplastic mucosa in spite of the Sox2 expression. Cdx2 increased the transcriptional activity of the Sox2 gene, and Sox2 increased the transcriptional activity of the Muc5Ac gene, which was reduced by cotransfecion of Cdx2 together with Sox2 in the human gastric carcinoma cell line AGS. In conclusion, Sox2 expression is maintained while gastric phenotype is completely lost in the intestinal metaplastic mucosa of Cdx2-transgenic mouse stomach. (C) 2010 International Society of Differentiation. Published by Elsevier Ltd. All rights reserved.
  • Hirotsugu Sakamoto, Hiroyuki Mutoh, Kentaro Sugano
    SCANDINAVIAN JOURNAL OF GASTROENTEROLOGY 45 11 1273 - 1280 2010年11月 [査読有り][通常論文]
     
    Objective. Cdx2 is expressed in human intestinal metaplastic mucosa and induces intestinal metaplastic mucosa in Cdx2-transgenic mouse stomach. Claudin-2 is a structural component of tight junctions in the intestine and Cdx2 activates the Claudin-2 promoter in the human intestinal epithelial cell line Caco-2. Our aim is to evaluate the expression of Claudin-2 in intestinal metaplastic mucosa of Cdx2-transgenic mouse stomach. Material and methods. The Claudin-2 expression in the normal gastric mucosa and normal intestinal mucosa of wild type mice and the intestinal metaplastic mucosa of Cdx2-transgenic mice was analyzed by immunohistochemistry, Western blotting and quantitative real-time PCR (qRT-PCR). Results. Claudin-2 was expressed in the base of the glands in intestine and intestinal metaplasia while it was not expressed in the body of stomach. Claudin-2 expression was found in the antrum of stomach, while it was weaker than that in the intestine and the intestinal metaplasia. Claudin-2 was also detected in intestinal metaplasia, colon and ileum by both Western blotting and qRT-PCR while it was not detected in gastric body. Conclusion. These results suggest that Cdx2 plays an important role in the expression of Claudin-2 in vivo.
  • Hiroyuki Mutoh, Miho Sashikawa, Hiroko Hayakawa, Kentaro Sugano
    CANCER SCIENCE 101 8 1783 - 1789 2010年08月 [査読有り][通常論文]
     
    Helicobacter pylori (H. pylori) stimulates secretion of monocyte chemoattractant protein 1 (MCP-1) from gastric mucosa. Monocyte chemoattractant protein-1 (MCP-1) expression and macrophage infiltration are recognized in human gastric carcinoma. We have previously generated Cdx2-transgenic mice as model mice for intestinal metaplasia. Both chronic H. pylori-associated gastritis and Cdx2-transgenic mouse stomach develop intestinal metaplasia and finally gastric carcinoma. In this study we have directed our attention to MCP-1 expression in the intestinal metaplastic mucosa and the gastric carcinoma of Cdx2-transgenic mouse stomach. Quantitative real-time PCR was performed to determine MCP-1 and transforming growth factor-beta 1 (TGF-beta 1) mRNA expression levels and single- or double-label immunohistochemistry was used to evaluate the localization of MCP-1, TGF-beta type I receptor, and alpha-smooth muscle actin (alpha SMA). We determined that MCP-1 mRNA dramatically increased in the intestinal metaplastic mucosa and the gastric carcinoma of Cdx2-transgenic mouse stomach, compared with normal mouse stomach. Both MCP-1 and TGF-beta type I receptor were co-expressed in the alpha SMA-positive myofibroblasts of intestinal metaplastic mucosa and gastric carcinoma. Exogenous application of TGF-beta 1 increased MCP-1 mRNA expression levels in the intestinal metaplastic tissue. Furthermore, TGF-beta 1 was overexpressed and macrophage was strongly infiltrated in the gastric carcinoma. In conclusion, MCP-1 expression, which was stimulated by TGF-beta 1, was recognized in the TGF-beta type I receptor-expressing myofibroblasts of the intestinal metaplastic mucosa and the gastric carcinoma of Cdx2-transgenic mouse stomach. The present results suggest that intestinal metaplasia and gastric carcinoma themselves induce MCP-1 expression independently of H. pylori infection. (Cancer Sci 2010).
  • Hiroyuki Mutoh, Hiroko Hayakawa, Miho Sashikawa, Hirotsugu Sakamoto, Kentaro Sugano
    BIOCHEMICAL JOURNAL 427 3 423 - 434 2010年05月 [査読有り][通常論文]
     
    Shh (Sonic Hedgehog) is a morphogen involved in gastric fundic gland differentiation in the adult. Shh expression is reduced in Helicobacter pylori-associated intestinal metaplastic change of the gastric epithelium and mice that lack Shh show intestinal transformation of the gastric mucosa. Similarly, in the stomach of Cdx2 (caudal-type homeobox 2)-transgenic mice, the gastric mucosa is replaced by intestinal metaplastic mucosa. The aim of the present study was to use Cdx2-transgenic mice to investigate: (i) Shh expression in the intestinal metaplastic mucosa of the Cdx2-transgenic mouse stomach; and (ii) the relationship between Shh and Cdx2. We determined that Shh mRNA levels were dramatically reduced in the intestinal metaplastic mucosa of the Cdx2-transgenic mouse stomach compared with the normal (wild-type) mouse stomach. This was not due to hypermethylation of the Shh promoter, but instead we showed that Cdx2 directly bound to the TATA box region of the Shh promoter. Cdx2 also down-regulated transcription of the Shh gene in the human gastric carcinoma cell lines AGS, MKN45 and MKN74. In conclusion, Cdx2 reduced Shh expression by binding to the unmethylated Shh promoter in the intestinal metaplastic mucosa of Cdx2-transgenic mouse stomach.
  • Hirotsugu Sakamoto, Hiroyuki Mutoh, Kenichi Ido, Shin Satoh, Machio Kumagai, Hiroko Hayakawa, Kiichi Tamada, Kentaro Sugano
    HUMAN PATHOLOGY 40 12 1762 - 1767 2009年12月 [査読有り][通常論文]
     
    We reported previously that intestinal metaplasia in the gallbladder is strongly associated with expression of caudal-related homeobox transcription factor Cdx2. It has been documented that occult pancreatobiliary reflux, even in the absence of pancreaticobiliary maljunction, is associated with elevated risk of biliary malignancy. We ascertained the correlation between intestinal metaplasia in the gallbladder and occult pancreatobiliary reflux. In 196 patients with a normal pancreaticobiliary ductal arrangement who had undergone laparoscopic cholecystectomy, we performed intraoperative cholangiography and measured amylase levels in bile sampled from the gallbladder. The cutoff value for high cystic amylase was defined as a biliary amylase level higher than the normal upper limit of serum amylase (215 IU/L). We also retrospectively reviewed the cholecystectomized tissue specimens to investigate the presence of intestinal metaplasia and expression of Cdx2. Then, we explored the relationship between intestinal metaplasia in the gallbladder and occult choledocho-pancreatic reflux. Intestinal metaplasia was found in 16.8% (33/196) of the gallbladders. The prevalence of choledochopancreatic reflux revealed by intraoperative cholangiography was not significantly different between cases with intestinal metaplasia (5/33, 15.2%) and those without (25/163, 15.3%; P = .81). However, in cases with intestinal metaplasia, the rate of high cystic amylase (13/33, 39.4%) was significantly higher compared with cases without intestinal metaplasia (26/163, 16.0%, P = .005). In conclusion, intestinal metaplasia in the gallbladder is significantly correlated with high amylase levels in bile in patients with a morphologically normal pancreaticobiliary ductal arrangement. (C) 2009 Published by Elsevier Inc.
  • Hiroyuki Mutoh, Hiroko Hayakawa, Hirotsugu Sakamoto, Miho Sashikawa, Kentaro Sugano
    FEBS JOURNAL 276 20 5821 - 5831 2009年10月 [査読有り][通常論文]
     
    Cdx1 and Cdx2, which are transcription factors regulating normal intestinal development, have been studied as potential key molecules in the pathogenesis of the precancerous intestinal metaplasia of the human stomach. However, the regulation of Cdx1 expression in the intestinal metaplasia is poorly understood. Cdx2-expressing gastric mucosa of Cdx2-transgenic mouse stomach was replaced by intestinal metaplastic mucosa. The aim of this study was to investigate the following: (a) Cdx1 expression in the intestinal metaplastic mucosa of the Cdx2-transgenic mouse stomach; and (b) the relationship between Cdx1 and Cdx2. A mouse model of intestinal metaplasia, the Cdx2-transgenic mouse, was used to investigate Cdx1 gene expression by RT-PCR. DNA methylation pro. le analysis was performed by bisulfite sequencing, and the interaction of Cdx2 with the Cdx1 promoter was examined by chromatin immunoprecipitation assay, electrophoretic mobility shift assay, and luciferase reporter assays. Cdx2 mRNA was expressed in the Cdx2-transgenic mouse stomach. However, endogenous Cdx2 mRNA was not expressed in the intestinal metaplasia of the Cdx2-transgenic mouse stomach. On the other hand, endogenous Cdx1 mRNA and protein were expressed in the intestinal metaplasia of the Cdx2-transgenic mouse stomach. The Cdx1 promoter was unmethylated in the intestinal metaplasia of the Cdx2-transgenic mouse stomach. Chromatin immunoprecipitation assay and electrophoretic mobility shift assay showed that Cdx2 was bound to the Cdx1 promoter region in the intestinal metaplasia and the normal intestine. Cdx2 upregulated and siRNA-Cdx2 downregulated the transcriptional activity of the Cdx1 gene in the human gastric carcinoma cell lines AGS, MKN45, and MKN74. In conclusion, transgenic Cdx2 induced endogenous Cdx1 through the binding of Cdx2 to the unmethylated Cdx1 promoter region in the intestinal metaplasia of the Cdx2-transgenic mouse stomach.
  • Hiroyuki Mutoh, Hiroko Hayakawa, Hirotsugu Sakamoto, Kentaro Sugano
    JOURNAL OF GASTROENTEROLOGY 42 9 719 - 729 2007年09月 [査読有り][通常論文]
     
    Background. While cyclooxygenase-2 (COX-2) is not normally expressed by epithelial cells lining the human colon, COX-2 protein is aberrantly overexpressed in premalignant adenomatous polyps and carcinomas of the human colon. On the other hand, Cdx2 has been identified as a colonic tumor-suppressor gene, besides its role in cell differentiation. However, the relationship between CDX2 attenuation and COX-2 overexpression in colorectal carcinoma has not been established. Here, we investigated the mechanistic link between CDX2 downregulation and COX-2 upregulation. Methods. Gene expression was examined by immunoblotting, reverse transcription-polymerase chain reaction, and promoter analysis. Promoter transactivation was quantified by using a luciferase construct. DNA binding of nuclear factor-kappa B (NF-kappa B) was examined by electromobility shift analysis. Results. CDX2 decreased expression of COX-2 mRNA and protein at the transcriptional level in the human colon cancer Caco-2 cell line. Though p50/p65 NF-kappa B translocated into nucleus in the presence of CDX2, CDX2 interacted with p50/p65 NF-kappa B and impeded the formation of an NF-kappa B-DNA complex, required for promotion of Cox-2 transcription. Conclusion. The results indicate that CDX2 inhibits transcription of Cox-2 by interfering with the binding of NF-kappa B on the NF-kappa B binding site.
  • Hirotsugu Sakamoto, Hiroyuki Mutoh, Kenichi Ido, Kiichi Satoh, Hiroko Hayakawa, Kentaro Sugano
    HUMAN PATHOLOGY 38 1 66 - 71 2007年01月 [査読有り][通常論文]
     
    We previously reported a case of a human gallbladder with cholelithiasis consisting of intestinal metaplasia with the expression of caudal-related homeobox transcription factor (Cdx2). However, it is unclear how often intestinal metaplasia and Cdx2 expression occur in human, nontumorous gallbladders with cholelithiasis. We studied the incidence of intestinal metaplasia and Cdx2 expression in human gallbladders with cholelithiasis. Gallbladders were resected under laparoscopy from 103 patients with cholelithiasis between September 2003 and March 2005. The mean age of the patients was 59.6 +/- 15.0 years (range, 22-92 years). We retrospectively reviewed these cases to look for the presence of intestinal metaplasia and the expression of Cdx2. In addition, the characteristics of intestinal metaplasia were examined by immunostaining for Muc2, chromogranin A, and serotonin. Intestinal metaplasia was found in 11.7% (12/103) of the gallbladders with cholelithiasis. The mean ages of patients with and without intestinal metaplasia were 60.8 +/- 15.4 and 59.4 +/- 14.9 years, respectively. Cdx2, Muc2, chromogranin A, and serotonin were expressed in 91.7% (11/12), 91.7% (11/12), 83.3% (10/12), and 50.0% (6/12) in intestinal metaplastic mucosa, respectively. Only one case (1.1%) that expressed Cdx2 without intestinal metaplasia did not express Muc2, chromogranin A, and serotonin. We found that 10.7% (11/103) of nontumorous gallbladders resected because of cholelithiasis under laparoscopy revealed intestinal metaplasia with Cdx2 expression. (c) 2007 Elsevier Inc. All rights reserved.
  • Hiroto Kita, Yuichi Hikichi, Kouki Hikami, Koichi Tsuneyama, Zheng-Guo Cui, Hiroyuki Osawa, Hirohide Ohnishi, Hiroyuki Mutoh, Hiroko Hoshino, Christopher L. Bowlus, Hironori Yamamoto, Kentaro Sugano
    JOURNAL OF GASTROENTEROLOGY 41 11 1053 - 1063 2006年11月 [査読有り][通常論文]
     
    Background: Flat adenomas in the colon are associated with a relatively higher potential for malignancy. Distinct genes may be involved in the development of flat adenoma. The aim of this study was to profile gene expression changes in flat adenomas in the colon. Methods: A genomewide expression analysis was carried out by using flat adenoma and adjacent normal mucosa in the colon to detect differences in gene expression. Because the right and left colon have different embryonic origins, each sample was classified according to its location, and the gene expression levels between flat adenoma and adjacent normal mucosa were also compared among samples derived from the right or left colon. Results :A total of 180 genes were differentially expressed between flat adenoma and normal mucosa in the colon, including matrix metalloproteinase 7 (MMP7), cadherin 3 (CDH3), S100P, and dual oxidase 2 (DUOX2). In addition, a total of 89 and 49 genes were differentially expressed between flat adenoma and normal mucosa among the samples from the right and left colon, respectively. Subsequent quantitative real-time reverse transcriptase-polymerase chain reaction supported the reliability of the expression analysis. Immunohistochemical analysis confirmed differential CDH3 and MMP7 protein expression. Conclusions :This is the first report characterizing the genes differentially expressed in flat adenomas using a microarray analysis. Considerable differences in the gene expression profiles of flat adenomas also exist between the right and left colon. These data should lead to new insights into the pathogenesis of flat adenomas in the colon as well as to new therapeutic strategies.
  • H Mutoh, H Sakamoto, H Hayakawa, Y Arao, K Satoh, M Nokubi, K Sugano
    DIFFERENTIATION 74 6 313 - 321 2006年07月 [査読有り][通常論文]
     
    The basic helix-loop-helix transcription factor Math1, which is transiently expressed in proliferating neural precursors in multiple domains of the developing nervous system, is also related to the cell fate decision of enteroendocrine, goblet, and Paneth cells in the intestine. On the other hand, the transcription factor Cdx2, which is normally confined to intestinal epithelial cells, is related to the differentiation of these cells. Therefore, we investigated the relationship between Math1 and Cdx2 in intestinal epithelial cells. The Math1 and Cdx2 expressions in normal intestinal mucosa and intestinal metaplastic mucosa from mouse and human stomachs, as well as an intestinal crypt-derived cell line, were analyzed by immunohistochemistry, reverse transcription-polymerase chain reaction and Northern blotting, and Math1 enhancer element was analyzed by luciferase reporter assays. Math1-positive epithelial cells co-expressing Cdx2 were found in normal intestinal mucosa from humans and mice. Furthermore, Math1-producing epithelial cells that showed positive immunostaining for Cdx2 were also observed in intestinal metaplastic mucosa from human and Cdx2 transgenic mouse stomachs, although they were not detected in normal gastric mucosa of humans and mice. Expression of Cdx2 stimulated endogenous Math1 mRNA expression in the intestinal crypt-derived cell line IEC-6, corroborating observations in Cdx2-expressing intestinal metaplastic mucosa. Furthermore, expression of Cdx2 in IEC-6 cells conferred the ability to express a Math1 reporter gene containing a Math1 enhancer. Based on these results, we hypothesize that Cdx2 is involved in activating Math1 expression in intestinal epithelial cells.
  • H Osawa, H Kita, H Ohnishi, H Hoshino, H Mutoh, Y Ishino, E Watanabe, K Satoh, K Sugano
    GUT 55 2 152 - 157 2006年02月 [査読有り][通常論文]
     
    Background and aims: Gastric acid secretion is downregulated by Helicobacter pylori infection and upregulated after its eradication, but the mechanisms are still unclear. We examined the effects of H pylori eradication on the number of parietal cells and on expression of molecules functioning in acid secretion in the human gastric mucosa. Methods: We enrolled 111 consecutive men with chronic gastritis induced by H pylori. Biopsy specimens were endoscopically obtained before and 12 weeks after successful eradication of H pylori and parietal cell numbers were counted. mRNA expression levels of H+/K+-adenosine triphosphatase ( H+/K+-ATPase), anion exchanger 2, M3 muscarinic receptor, intrinsic factor, and interleukin 1 beta were determined with a real time reverse transcriptase-polymerase chain reaction method. The severity of gastric atrophy was evaluated using the serum pepsinogen I/II ratio. Results: No significant difference was observed in parietal cell numbers before and after H pylori eradication. Median mRNA expression levels of H+/K+-ATPase in the gastric mucosa increased 250-fold after H pylori eradication accompanied by attenuation of interleukin 1 beta. A large increase in H+/K+-ATPase expression was observed even in patients with severe atrophic gastritis. In contrast, fold increases in mRNA expression levels, including intrinsic factor, anion exchanger 2, and M3 muscarinic receptor, after eradication therapy, were limited to 1.4, 2.3, and 2.5 times, respectively. Conclusions: In the absence of alteration of parietal cell number, gastric H+/K+-ATPase mRNA expression was markedly restored after successful H pylori eradication, suggesting a central role for the restoration of H+/K+-ATPase expression in gastric acid secretion recovery after H pylori eradication.
  • Mutoh H
    Nihon rinsho. Japanese journal of clinical medicine 63 1367 - 1371 8 2005年08月 [査読有り][通常論文]
  • Mutoh H
    Nihon Shokakibyo Gakkai zasshi = The Japanese journal of gastro-enterology 102 986 - 993 8 2005年08月 [査読有り][通常論文]
  • H Osawa, H Kita, H Ohnishi, H Mutoh, Y Ishino, K Satoh, K Sugano
    ALIMENTARY PHARMACOLOGY & THERAPEUTICS 21 92 - 98 2005年06月 [査読有り][通常論文]
     
    Background: Helicobacter pylori infection prevents the occurrence of the tolerance phenomenon of Histamine-2 (H2) receptor antagonists. Gastro-esophageal reflux disease develops in some cases with the restoration of acid secretion after H. pylori eradication therapy. Aim: To clarify the mechanisms of H2 receptor restoration after the eradication of H. pylori on parietal cells. Methods: We enrolled 80 consecutive asymptomatic male patients with H. pylori infection, having chronic gastritis with or without the presence of peptic ulcers. Biopsy specimens from the greater curvatures at the mid-corpus of the stomach were obtained endoscopically from all subjects before and 12 weeks after the eradication of H. pylori. Degrees of gastric atrophy were evaluated by serum pepsinogen levels. The amounts of mRNA expression of H2 receptor were evaluated in each subject's gastric mucosa by real time reverse transcriptase-polymerase chain reaction (RT-PCR). Results: H2 receptor mRNA expression levels significantly correlated with serum pepsinogens I and 11 ratios. The expression level of H2 receptor mRNA was lower in subjects with hypergastrinemia. The median expression level of H2 receptor after H. pylori eradication was threefold greater than prior to treatment. In addition, its restoration became more pronounced in subjects with severe gastric atrophy. However, a comparatively low restoration of H2 receptor mRNA was found in subjects with hypergastrinemia. Conclusions: H2 receptor mRNA levels decrease with the progression of gastric atrophy induced by H. pylori infection, and arc restored after H. pylori eradication. Such expression levels of H2 receptor may explain a part of the tolerance phenomenon to H2 receptor antagonists.
  • H Mutoh, S Sakurai, K Satoh, H Osawa, T Tomiyama, H Kita, T Yoshida, K Tamada, H Yamamoto, N Isoda, K Ido, K Sugano
    GUT 54 1 33 - 39 2005年01月 [査読有り][通常論文]
     
    Background and aims: In the progression of chronic gastritis, gastric mucosal cells deviate from the normal pathway of gastric differentiation to an intestinal phenotype which is closely related to gastric carcinoma. However, to date, it has not been elucidated whether the intestinal metaplasia is merely a change in the epithelium or whether the underlying mesenchyme also changes from gastric type to intestinal type. We have investigated the relationship between intestinal metaplasia and the pericryptal fibroblast sheath (PCFS) in the mesenchyme. In addition, we also examined PCFS in gastric carcinoma. Methods: We determined the existence of PCFS in the intestinal metaplastic mucosa and carcinoma of both human and Cdx2 transgenic mouse stomach. PCFS was determined using the antibody against alpha-smooth muscle actin and electron microscopic observations. Results: PCFS formed an almost complete layer around the small and large intestinal crypts while it did not exist around the normal gastric glands in both mice and humans. PCFS was seen around the glands of intestinal metaplastic mucosa in both Cdx2 transgenic mouse and human stomachs. However, PCFS was virtually absent in the intestinal-type gastric adenocarcinoma area. Conclusion: We successfully demonstrated that the epithelium as well as the mesenchyme changed from the gastric type to the intestinal type in intestinal metaplasia and that PCFS disappeared in intestinal-type gastric carcinoma.
  • H Mutoh, K Satoh, H Kita, H Sakamoto, H Hayakawa, H Yamamoto, N Isoda, K Tamada, K Ido, K Sugano
    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY 49 7 867 - 871 2005年 [査読有り][通常論文]
     
    Many transcription factors are involved in the molecular control of intestinal epithelial cell differentiation. We report in this study that the transcription factor Cdx2 functions to define absorptive enterocytes during intestinal epithelial differentiation. Cdx2 is expressed in the villi of the normal small intestine. Intestinal metaplasia, which expresses Cdx2, occurs as a pathological condition in gastric mucosa. We have previously established Cdx2transgenic mice expressing Cdx2 exclusively in the gastric epithelium. In this study using Cdx2 transgenic mice, we show that Cdx2 plays a key role in the differentiation of intestinal absorptive enterocytes. The gastric mucosa of Cdx2 transgenic mice was morphologically completely changed into intestinal metaplastic mucosa. Absorptive enterocytes had microvilli which were observed by electron microscope. The intestinal metaplastic mucosa of Cdx2 transgenic mice expressed sucrase and peptide transporter PepT1. Disaccharidase and leucine aminopeptidase activities were observed in the intestinal metaplastic mucosa. Glucose and amino acids were absorbed from Cdx2 transgenic mouse stomach with intestinal metaplasia. Finally we generated mice whose intestine was extensively excised. Cdx2 transgenic mice with intestinal metaplasia survived even after extensive intestinal excision. We successfully demonstrated that Cdx2 induced not only morphological but also functional absorptive enterocytes in the intestinal metaplastic mucosa in vivo. Our results suggest that Cdx2 is necessary and sufficient by itself to specify the development of intestinal absorptive enterocytes, whereas other factors which are expressed in the small intestine are not always necessary for the differentiation of functional absorptive enterocytes.
  • Y Satoh, H Kita, K Kihira, H Mutoh, H Osawa, K Satoh, K Ido, Y Sakata, K Sugano
    AMERICAN JOURNAL OF GASTROENTEROLOGY 99 12 2495 - 2498 2004年12月 [査読有り][通常論文]
     
    Although the association between gastrointestinal angiodysplasia and von Willebrand's disease has been suggested, molecular mechanisms involved in the formation of angiodysplasia in patients with von Willebrand's disease remained undetermined. We examined exon 28 of the von Willebrand factor gene in a patient with both von Willebrand's disease and recurrent bleeding from angiodysplasia in the duodenum as well as his father's, and found a point mutation, C 39:16-->T (amino acid substitution; Arg 543-->Trp), in the A1 domain of the von Willebrand factor gene. This mutation was identical with a previously reported mutation in a patient with von Willebrand's disease complicated with gastrointestinal angiodysplasia.
  • H Mutoh, S Sakurai, K Satoh, K Tamada, H Kita, H Osawa, T Tomiyama, Y Sato, H Yamamoto, N Isoda, T Yoshida, K Ido, K Sugano
    CANCER RESEARCH 64 21 7740 - 7747 2004年11月 [査読有り][通常論文]
     
    In the progression of chronic gastritis, gastric mucosal cells deviate from the normal pathway of gastric differentiation to an intestinal phenotype. Many epidemiologic studies have found an association between the formation of intestinal metaplasia and the development of gastric carcinoma. However, there is no direct evidence that shows intestinal metaplasia is a precursor lesion of gastric carcinoma, to date. We periodically examined the intestinal metaplastic mucosa of Cdx2-transgenic mice we have previously generated. Gastric polyps developed from intestinal metaplastic mucosa in all stomachs of Cdx2-transgenic mice examined. These gastric polyps consisted of intestinal-type adenocarcinoma that invaded the submucosa and muscularis propria and occasionally spread into the subserosa. p53 and APC gene mutations were recognized in the adenocarcinomas. The participation of APC and p53 gene mutations in gastric carcinogenesis from the intestinal metaplasia was verified by the Cdx2-transgenic mice, carrying Apc(Min) mutation or p53 deficiency, that developed gastric polyps much earlier than Cdx2 alone. We successfully showed that long-term intestinal metaplasia induces invasive gastric carcinoma. These results indicate that intestinal metaplasia itself plays a significant role in the genesis and progression of gastric carcinoma.
  • H Mutoh, S Sakurai, K Satoh, H Osawa, Y Hakamata, T Takeuchi, K Sugano
    GUT 53 10 1416 - 1423 2004年10月 [査読有り][通常論文]
     
    Background and aims: Gastric intestinal metaplasia, which is mainly induced by Helicobacter pylori infection, is thought to be a precancerous lesion of gastric adenocarcinoma. Intestinal metaplastic mucosa expresses intestine specific homeobox genes, Cdx1 and Cdx2, in the human gastric mucosa. We and others have reported that ectopic expression of Cdx2 in the gastric epithelium generates intestinal metaplasia in the transgenic mouse model. Methods: To clarify the differences in the roles of Cdx1 and Cdx2 in intestinal metaplasia, we generated transgenic mice expressing Cdx1 in the gastric mucosa and compared Cdx1 induced gastric mucosal morphological changes with Cdx2 induced intestinal metaplasia. Results: The gastric mucosa in Cdx1 transgenic mice was completely replaced by intestinal metaplastic mucosa, consisting of all four intestinal epithelial cell types: absorptive enterocytes, goblet, enteroendocrine, and Paneth cells. Paneth cells, which were not recognised in Cdx2 transgenic mice, were in the upper portion of the intestinal metaplastic mucosa. Pseudopyloric gland metaplasia, which was induced in Cdx2 transgenic mice, was not recognised in Cdx1 transgenic mice. Proliferating cell nuclear antigen ( PCNA) positive cells were diffusely scattered in Cdx1 induced intestinal metaplastic mucosa while PCNA positive cells in Cdx2 induced intestinal metaplastic mucosa were in the base of the metaplastic mucosa. Intestinal metaplastic mucosa of Cdx1 transgenic mouse stomach was significantly thicker than that of wildtype or Cdx2 transgenic mouse stomach. Conclusions: We have confirmed that Cdx1 induced gastric intestinal metaplasia but that it differed from Cdx2 induced intestinal metaplasia in differentiation, structure, and proliferation.
  • H Osawa, H Kita, K Satoh, H Ohnishi, Y Kaneko, H Mutoh, K Tamada, K Ido, K Sugano
    AMERICAN JOURNAL OF SURGICAL PATHOLOGY 28 9 1253 - 1254 2004年09月 [査読有り][通常論文]
  • K Satoh, H Kawata, K Tokumaru, Y Kumakura, Y Ishino, S Kawakami, K Inoue, T Kojima, Y Satoh, H Mutoh, K Kihira, K Sugano
    DIGESTIVE AND LIVER DISEASE 35 2 78 - 84 2003年02月 [査読有り][通常論文]
     
    Background. Change in apoptosis in gastric glands after eradication of Helicobacter pylori has never been reported. Aims. The purpose of this paper is to investigate the change in apoptosis in gastric glands after eradication of Heliobacter pylori. Patients and methods. We studied 23 Heliobacter pylori-positive patients with duodenal and gastric ulcers, who were monitored for 6-12 months after eradication, and eight controls. Biopsies were taken from the antrum and body. Apoptosis was evaluated immunohistochemically using anti-single stranded DNA antibody. Apoptotic index was calculated by counting immunostained cells in surface epithelial and glandular cells. Results. In the surface epithelium, Apoptotic indexes were significantly higher in patients than in controls. In the upper portion of fundic glands, apoptotic indexes were significantly higher in patients with gastric ulcers (14.2% (9.3, 17.8)) (median (1st quartile, 3rd quartile)) than in controls (8.0% (2.0, 9.0), p<0.01) and decreased significantly after eradication (3.4% (2.0, 5.3)), p<0.01). In pyloric glands, apoptotic indexes were no different between patients and controls. In the lower portion of fundic glands, apoptotic indexes were very low, both in patients and in controls. Conclusions. Our results showed that apoptosis, not only of surface epithelial cells but also of glandular cells in the upper portion of fundic glands, increased in Heliobacter pylori-positive patients with gastric ulcers and decreased to normal levels after eradication of Heliobacter pylori. (C) 2003 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Science Ireland Ltd. All rights reserved.
  • A Eda, H Osawa, K Satoh, Yanaka, I, K Kihira, Y Ishino, H Mutoh, K Sugano
    JOURNAL OF GASTROENTEROLOGY 38 1 14 - 22 2003年01月 [査読有り][通常論文]
     
    Background. There have been no detailed reports directly comparing the expression of CDX1 with that of CDX2 in the inflammatory esophageal mucosa and Barrett's epithelium. The present study was designed to examine the expression of CDX 1/2 in inflammatory esophageal mucosa with or without Barrett's epithelium. Methods. The expression of CDX1/2 genes was analyzed using the reverse transcriptase-polymerase chain reaction (RT-PCR) in 34 human esophageal biopsy specimens, and CDX2 expression was also evaluated immunohistochemically, using anti-human CDX2 monoclonal antibody. The biopsy specimens for RNA extraction were taken endoscopically from esophageal mucosa with mucosal break due to gastroesophageal reflux disease (GERD), Barrett's epithelium, and normal epithelium. The expressions of mucin markers (MUC2) and intestine-specific genes (sucrase-isomaltase, human defensin-5, alkaline phosphatase) were also comparatively analyzed. Results. CDX1/2 expression was not found in the normal esophageal mucosa. The prevalence of CDX1/2 mRNA expression was significantly higher in the mucosa with Barrett's epithelium than in the mucosa without Barrett's epithelium. It is noteworthy, however, that the CDX2 mRNA expression was initiated at the stage of esophagitis, when neither CDX1 nor intestine-specific genes had emerged yet. In contrast to CDX2, CDX1 was expressed only in Barrett's epithelium. Immunohistochemical study demonstrated strong and extensive nuclear immunoreactivity for CDX2 in Barrett's epithelium. Furthermore, fine granular cytoplasmic staining was also observed in the cytoplasm in Barrett's epithelium, as well as in inflammatory esophageal mucosa. Conclusions. We report here, for the first time, that CDX2 is expressed in patients with Barrett's epithelium and inflammatory esophageal mucosa. These findings imply that the expression of CDX2 may be an early event leading to the development of Barrett's esophagus.
  • K Satoh, H Mutoh, A Eda, Yanaka, I, H Osawa, S Honda, H Kawata, K Kihira, K Sugano
    HELICOBACTER 7 3 192 - 198 2002年06月 [査読有り][通常論文]
     
    Background, The intestine-specific transcription factor CDX2 plays an important role in differentiation and maintenance of intestinal epithelial cells. Development and progression of intestinal metaplasia (IM) in the stomach is closely associated with Helicobacter pylori -gastritis. We investigated expression of CDX2 protein in the gastric mucosa with and without IM before and after eradication of H. pylori . Materials and Methods. The subjects comprised five normal controls and 29 H. pylori -positive patients (15 with antral IM and 14 without IM), who were followed for 12 months after eradication of H. pylori . Biopsies were taken from the greater curvatures of the antrum and middle body. Expression of CDX2 was evaluated immunohistochemically using anti-CDX2 antibody. Results. CDX2 expression was not found in controls. Strong nuclear staining was observed extensively in IM, but rarely in the gastric epithelium, except for the focal area in only four antral biopsies (three with and one without IM). Fine granular cytoplasmic staining was also observed in the perinuclear regions of IM and the gastric epithelial cells in half of the patients. In 13 of the 15 patients with IM, IM did not regress after eradication of H. pylori , and the extent of nuclear staining in IM did not change. The extent of cytoplasmic staining did not change either. Conclusions. Our results indicate that CDX2 expression in the gastric mucosa is found in patients with chronic gastritis and is closely associated with IM. CDX2 expression in IM or the gastric epithelial cells did not disappear after eradication of H. pylori .
  • H Mutoh, Y Hakamata, K Sato, A Eda, Yanaka, I, S Honda, H Osawa, Y Kaneko, K Sugano
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 294 2 470 - 479 2002年06月 [査読有り][通常論文]
     
    Gastric intestinal metaplasia occurs as a pathological condition in the gastric mucosa. To clarify how an intestine-specific homeobox gene, Cdx2, affects the morphogenesis of gastric mucosa, we generated transgenic mice expressing Cdx2 in parietal cells. Until Day 18 after birth, the number of parietal cells inthegastric mucosa of transgenic mice was the same as for their normal littermates. However, at Day 19, we detected several glands in which parietal cells disappeared and the proliferating zone moved from the isthmus to the base of the glands. Thereafter, parietal cells decreased gradually and disappeared at Day 37. All of the gastric mucosal cells, except for enterochromaffin-like (ECL) cells, were completely replaced by intestinal metaplasia, consisting of goblet cells, enteroendocrine cells, and absorptive cells expressing alkaline phosphatase. Pseudopyloric gland metaplasia was also formed, The transgenic mouse is a very useful model for clarifying physiological differentiation of gastric and intestinal cell lineages and analyzing the molecular events from intestinal metaplasia. to adenocarcinoma. (C) 2002 Elsevier Science (USA). All rights reserved.
  • C Ratineau, MW Petry, H Mutoh, AB Leiter
    JOURNAL OF BIOLOGICAL CHEMISTRY 277 11 8847 - 8853 2002年03月 [査読有り][通常論文]
     
    Expression of the hormone secretin in enteroendocrine cells is restricted to the nondividing villus compartment of the intestine, implying that terminal differentiation is linked to cell cycle arrest and that differentiation is repressed in actively proliferating cells. We have shown previously that the basic helix-loop-helix protein, BETA2/NeuroD, induces cell cycle withdrawal in addition to increasing secretin gene expression. A number of transcription factors important for differentiation are repressed by D cyclins. Repression by D cyclins appears to be independent of its effects on the cell cycle. We show that cyclin D1 represses BETA2/NeuroD-dependent transcription of the secretin gene. Examination of cyclin box mutants shows that repression is unrelated to Cdk4 activation. Although cyclin D1 and BETA2 associate in vivo, they do not directly interact. Cyclin D1 may be recruited to BETA2 by binding to the C-terminal domain of the p300 coactivator, downstream from the BETA2-binding site. In the small intestine, cyclin D1 expression occurs only in the actively proliferating crypts of Lieberkuhn but not in villi. Thus repression by cyclin D I may serve to prevent secretin gene transcription from occurring in relatively immature epithelial progenitor cells.
  • A Eda, H Osawa, Yanaka, I, K Satoh, H Mutoh, K Kihira, K Sugano
    JOURNAL OF GASTROENTEROLOGY 37 2 94 - 100 2002年02月 [査読有り][通常論文]
     
    The CDX1 and CDX2 genes are intestinal transcription factors that may be involved in the regulation of proliferation and differentiation of intestinal epithelial cells. There have been no detailed reports directly comparing the expression of CDX1 with that of CDX2 in chronic gastritis and intestinal metaplasia. Accordingly, we examined the expression of CDX1/2 and its association with the expression of other intestinal metaplasia-associated genes during the development of intestinal metaplasia. Methods. The expression of CDX1/2 genes was analyzed, using the reverse transcriptase-polymerase chain reaction, in 44 human gastric tissue samples obtained endoscopically. The expressions of mucin markers (MUC2, MUC5AC), intestine-specific genes (sucrase-isomaltase, human defensin-5, alkaline phosphatase). a gene marker for fundic gland area (H+/K(+)ATPase beta subunit), and a gene for entire gastric glands (pepsinogen C) were also comparatively analyzed. Results. There was no expression of CDX1/2 in gastric mucosa not infected by Helicobacter pylori. The prevalence of CDX1 mRNA expression was significantly higher in mucosa with intestinal metaplasia than in mucosa without intestinal metaplasia. It is noteworthy that CDX2 was expressed in the antral and fundic mucosa in the absence of the expression of CDX1 and gene markers for intestinal metaplasia. Conclusions. The expression of CDX2 precedes those of CDX1, sucrase-isomaltase, other intestine-specific genes (human defensin-5, alkaline phosphatase), and MUC2 during the progression of intestinal metaplasia. These findings imply that the expression of CDX2 may trigger the initiation and development of intestinal metaplasia.
  • Mutoh H, Sugano K
    Nihon rinsho. Japanese journal of clinical medicine 60 Suppl 2 252 - 255 2002年02月 [査読有り][通常論文]
  • Mutoh H, Sugano K
    Nihon rinsho. Japanese journal of clinical medicine 60 Suppl 1 151 - 160 2002年01月 [査読有り][通常論文]
  • J Wang, K Tahara, Y Hakamata, H Mutoh, T Murakami, M Takahashi, M Kusama, E Kobayashi
    MICROSURGERY 22 8 371 - 377 2002年 [査読有り][通常論文]
     
    Auxiliary partial liver transplantation (APLT) is beneficial for fulminant liver failure when there is potential for recovery of the diseased liver. However, the impact of host hepatectomy on regeneration of the grafted liver is unclear. In this study, we modified a previous rat model of auxiliary whole liver transplantation without portal vein reconstruction, and studied the effect of host hepatectomy on regeneration of the cut liver graft. Thirty percent of the liver was heterotopically transplanted, to connect the recipient's left renal artery and vein with the graft's aortic cuff of the hepatic artery and inferior vena cava, respectively, using a cuff technique; 30% of the recipient liver then was cut. The control group was left intact. The liver grafts were weighed preoperatively and 2 weeks postoperatively. This procedure prevented congestion of the graft liver and achieved a high success rate, even when performed by a surgeon who was relatively inexperienced with the technique. The weight of the grafted liver in the host hepatectomized group significantly increased (P < 0.05) compared with that of the control group. We developed an experimental model of APLT and reviewed the literature on rat heterotopic liver transplantation, and compared the surgical techniques. (C) 2002 Wiley-Liss, Inc.
  • K Satoh, K Kihira, H Kawata, K Tokumaru, Y Kumakura, Y Ishino, S Kawakami, K Inoue, T Kojima, Y Satoh, H Mutoh, K Sugano
    HELICOBACTER 6 1 31 - 36 2001年03月 [査読有り][通常論文]
     
    Background. Accumulation of p53 has been recognized in the gastric mucosa infected with Helicobacter pylori. We investigated the prevalence of p53-positive cells in the gastric mucosa before and one month after eradication of H. pylori and the relationship between p53 positivity and inflammation and cell proliferation. Methods. The subjects included 24 H. pylori-posit ive patients. They achieved eradication one month after anti-H. pylori therapy. Biopsies were taken from the greater curvatures of the antrum and middle body. H. pylori status was assessed using culture and tissue section (Giemsa stain). Serial sections were used for examination of gastritis (hematoxylin and eosin stain) and for immunostaining of p53, Ki-67 and myeloperoxidase (MPO). p53 index and Ki-67 labeling index (LI) were calculated by counting p53-positive and Ki-67-positive cells in the entire gastric pits longitudinally sectioned and expressing them as a percentage of the total cells in a gastric pit. In the neck regions with and without p53-positive cells, polymorphonuclear leukocytes (PMNs) were counted in the corresponding area (/50x50 mum(2)) of the sections stained both with p53 and MPO. Results. p53-positive cells decreased significantly after eradication of H. pylori. Before eradication, the number of PMNs was significantly higher in the neck regions with p53-positive cells than in those without. Conclusions. In the gastric mucosa infected with H. pylori, p53-positive cells were found in the neck region infiltrated with PMNs. p53 expression decreased significantly one month after eradication of H. pylori.
  • H Mutoh, C Ratineau, S Ray, AB Leiter
    ALIMENTARY PHARMACOLOGY & THERAPEUTICS 14 170 - 175 2000年04月 [査読有り][通常論文]
     
    Secretin-producing enteroendocrine cells arise from a multipotential endocrine progenitor in the crypts of the small intestine. As these cells migrate up the crypt-villus axis, they produce secretin and stop dividing as they terminally differentiate and die. Transcription of the secretin gene is controlled by a complex enhancer binding to multiple transcription factors. The basic helix-loop-helix protein, BETA2, binds to an E box sequence and associates with the p300 coactivator to activate transcription of the secretin gene. Basic helix-loop-helix proteins appear to play a pivotal role in I-he control of cellular differentiation. BETA2 induces cell cycle arrest and apoptosis in addition to activating secretin gene expression. Thus BETA2 may function as a master regulatory gene to coordinate terminal differentiation of secretin cells.
  • H Mutoh, FJ Naya, MJ Tsai, AB Leiter
    GENES & DEVELOPMENT 12 6 820 - 830 1998年03月 [査読有り][通常論文]
     
    The major epithelial cell types lining the intestine comprise a perpetually self-renewing population of cells that differentiate continuously from a stem cell in the intestinal crypts. Secretin-producing enteroendocrine cells represent a nondividing subpopulation of intestinal epithelial cells, suggesting that expression of the hormone is coordinated with cell cycle arrest during the differentiation of this cell lineage. Here we report that the basic helix-loop-helix protein BETA2 associates functionally with the coactivator, p300 to activate transcription of the secretin gene as well as the gene encoding the cyclin-dependent kinase inhibitor p21. Overexpression of BETA2 in cell lines induces both cell cycle arrest and apoptosis suggesting that BETA2 may regulate proliferation of secretin cells. Consistent with this role, we observed both reentry of normally quiescent cells into the cell cycle and disrupted cell number regulation in the small intestine of BETA2 null mice. Thus, BETA2 may function to coordinate transcriptional activation of the secretin gene, cell cycle arrest, and cell number regulation, providing one of the first examples of a transcription factor that controls terminal differentiation of cells in the intestinal epithelium.
  • FJ Naya, HP Huang, YH Qiu, H Mutoh, FJ DeMayo, AB Leiter, MJ Tsai
    GENES & DEVELOPMENT 11 18 2323 - 2334 1997年09月 [査読有り][通常論文]
     
    Candidate transcription factors involved in pancreatic endocrine development have been isolated using insulin gene regulation as a paradigm. The cell-type restricted basic helix-loop-helix (bHLH) gene, BETA2/NeuroD, expressed in pancreatic endocrine cells, the intestine, and the brain, activates insulin gene transcription and can induce neurons to differentiate. To understand the importance of BETA2 in pancreatic endocrine cell differentiation, mice lacking a functional BETA2 gene were generated by gene targeting experiments. Mice carrying a targeted disruption of the BETA2 gene developed severe diabetes and died perinatally. Homozygous BETA2 null mice had a striking reduction in the number of insulin-producing beta cells and failed to develop mature islets. Islet morphogenesis appeared to be arrested between E14.5 and E17.5, a period characterized by major expansion of the beta cell population. The presence of severe diabetes in these mice suggests that proper islet structure plays an important role in blood glucose homeostasis. In addition, secretin-and cholecystokinin-producing enteroendocrine cells failed to develop in the absence of BETA2. The absence of these two pancreatic secretagogs may explain the abnormal cellular polarity and inability to secrete zymogen granules in pancreatic acinar exocrine cells. The nervous system appeared to develop normally, despite abundant expression of BETA2 in differentiating neurons. Thus, BETA2 is critical for the normal development of several specialized cell types arising from the gut endoderm.
  • H Mutoh, BP Fung, FJ Naya, MJ Tsai, J Nishitani, AB Leiter
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 94 8 3560 - 3564 1997年04月 [査読有り][通常論文]
     
    The gene encoding the hormone secretin is expressed only in enteroendocrine S cells and insulin-producing pancreatic beta cells during development. A 120-bp enhancer directs cell-specific expression of the rat secretin gene in secretin-expressing cells. The enhancer includes an E-box sequence, CAGCTG, which is important for transcriptional activity. To further characterize the role of the E box, a consensus binding site for basic helix-loop-helix (bHLH) proteins, we have examined factors that interact with this element in the secretin gene. The results suggest that transcription is activated by a recently isolated tissue specific bHLH protein, BETA2, heterodimerized to the ubiquitously expressed bHLH proteins, Pan 1 and Pan 2, the rodent homologues of E47 and E12. The importance of BETA2 for transcriptional activation of secretin is further illustrated by antisense experiments inhibiting BETA2 expression in secretin-producing cell lines, which resulted in the inhibition of most E box-dependent transcription. Expression of BETA2 in a nonendocrine cell line conferred the ability to express secretin-reporter genes that are transcribed at minimal levels in the absence of BETA2. Secretin producing enteroendocrine cells in the murine small intestine showed specific immunostaining with BETA2 antibodies, corroborating observations in cell lines. Thus BETA2 is to our knowledge the first transcription factor identified that specifically activates cell type-specific expression of an intestinal hormone gene.
  • Shimizu T, Mutoh H
    Advances in experimental medicine and biology 407 197 - 204 1997年 [査読有り][通常論文]
  • Shimizu T, Mutoh H, Kato S
    Advances in experimental medicine and biology 416 79 - 84 1996年 [査読有り][通常論文]
  • Sato S, Kume K, Takan T, Mutoh H, Taketani Y, Shimizu T
    Advances in experimental medicine and biology 416 95 - 100 1996年 [査読有り][通常論文]
  • H Mutoh, T Fukuda, T Kitamaoto, S Masushige, H Sasaki, T Shimizu, S Kato
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 93 2 774 - 779 1996年01月 [査読有り][通常論文]
     
    We have studied the effects of retinoic acid (RA) and thyroid hormone (3,3',5-triiodothyronine; T-3) on platelet-activating factor receptor (PAFR) gene expression in intact rats and the ability of two human PAFR gene promoters (PAFR promoters 1 and 2) to generate two transcripts (PAFR transcripts 1 and 2), Northern blotting showed that RA and T-3 regulated PAFR gene expression only in rat tissues that express PAFR transcript 2. Functional analysis of the human PAFR promoter 2 revealed that responsiveness to RA and T-3 was conferred through a 24-bp element [PAFR-hormone response element (HRE)] located from -67 to -34 bp of the transcription start site, whereas PAFR promoter 1 did not respond to these hormones, The PAFR-HRE is composed of three direct repeated TGACCT-like hexamer motifs with 2- and 4-bp spaces, and the two upstream and two downstream motifs were identified as response elements for RA and T-3. Thus, the PAF-PAFR pathway is regulated by the PAFR level altered by a tissue-specific response to RA and T-3 through the PAFR-HRE of the PAFR promoter 2.
  • H Mutoh, H Hiraishi, S Ota, A Terano, K Ogura, KJ Ivey, T Sugimoto
    DIGESTIVE DISEASES AND SCIENCES 40 12 2704 - 2711 1995年12月 [査読有り][通常論文]
     
    The current study investigated whether metal ions were cytoprotective against ethanol-induced injury to cultured rat gastric mucosal cells in vitro. Secondly, the relationships. between oxygen free radicals and cytoprotection by metal ions were examined. Cultured cells exposed to ethanol produced superoxide anion, as assessed by reduction of cytochrome c, in a time-related fashion, and the production of superoxide anion increased dose-dependently as the concentration of ethanol increased. Cellular damage increased proportionately to the production of superoxide anion. ZnCl2, AlCl3, CoCl2, CuCl2, and CdCl2 significantly diminished ethanol-induced injury dose-dependently. All of the agents studied decreased the reduction of cytochrome c in ethanol-induced damage dose-dependently. These results led to the conclusions that: (1) cultured rat gastric mucosal cells exposed to ethanol generate oxygen free radicals; (2) the production of oxygen free radicals is closely linked with ethanol-induced damage to the cells; and (3) metal ions decrease ethanol-induced gastric mucosal cell damage in vitro. Metal ions protect cultured rat gastric mucosal cells from ethanol-induced damage in which oxygen free radicals participate.
  • T IZUMI, T TAKANO, H BITO, M NAKAMURA, H MUTOH, Z HONDA, T SHINIZU
    JOURNAL OF LIPID MEDIATORS AND CELL SIGNALLING 12 2-3 429 - 442 1995年10月 [査読有り][通常論文]
  • M MINAMI, H MUTOH, N OHISHI, ZI HONDA, H BITO, T SHIMIZU
    GENE 161 2 249 - 251 1995年08月 [査読有り][通常論文]
     
    The guinea-pig leukotriene A(4) hydrolase (LTA(4)H)-encoding cDNA was isolated from a guinea-pig lung cDNA library by cross-hybridization using a human probe. The deduced amino acid (aa) sequence consists of 611 aa (68 756 Da) and contains all twelve internal peptide and N-terminal sequences determined from the purified enzyme from guinea-pig intestine. The aa identity of the guinea-pig enzyme with its human, mouse and rat counterparts was 92.9, 90.5 and 90.4%, respectively. The previously characterized zinc-binding motif and a putative active site were highly conserved, supporting the aminopeptidase activity described for this enzyme. RNA blot analysis demonstrated ubiquitous expression of the LTA(4)H mRNA.
  • H MUTOH, S OTA, H HIRAISHI, KJ IVEY, A TERANO, T SUGIMOTO
    DIGESTIVE DISEASES AND SCIENCES 40 4 872 - 878 1995年04月 [査読有り][通常論文]
     
    In cultured gastric mucosal cells, we investigated whether: (1) adaptive cytoprotection was associated with stimulation of endogenous prostaglandin synthesis; (2) prostaglandins given exogenously were cytoprotective against ethanol-induced gastric mucosal cell damage; and (3) a relationship existed between cytoprotection and mucus release. Cytolysis was quantified by measuring Cr-51 release from prelabeled cells. Mucus release was determined by measurement of [H-3]glucosamine release. Concentrations of ethanol >12% caused cell damage and increased Cr-51 release dose dependently. Pretreatment with low concentrations of ethanol (0.5-1.5%) decreased ethanol-induced Cr-51 release, but also decreased prostaglandin E(2) synthesis. Prostaglandin E(2) and 16,16-dimethyl prostaglandin E(2) given exogenously were cytoprotective against ethanol-induced gastric mucosal cell damage. Treatment with low concentrations of ethanol (1.5%) increased mucus release from cultured gastric mucosal cells. However, prostaglandin E(2) and 16,16-dimethyl prostaglandin E(2) did not affect mucus release. We conclude that in cultured gastric mucus-producing cells: (1) adaptive cytoprotection occurs without stimulation of endogenous prostaglandin synthesis but with increase in mucus release; and (2) exogenous prostaglandins are cytoprotective against ethanol-induced gastric mucosal cell damage without stimulating mucus release in vitro. We postulate that adaptive cytoprotection in cultured gastric mucus-producing cells is not mediated by prostaglandin, but by mucus released in response to a mild irritant.
  • Izumi T, Honda Z, Mutoh H, Kume K, Shimizu T
    Advances in prostaglandin, thromboxane, and leukotriene research 23 461 - 466 1995年 [査読有り][通常論文]
  • H MUTOH, K KUME, S SATO, S KATO, T SHIMIZU
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 205 2 1130 - 1136 1994年12月 [査読有り][通常論文]
     
    We found that the expression of human platelet-activating factor receptor (PAFR) gene is differentially regulated by estrogen and TGF-beta 1. Primer extension analysis revealed that the levels of the PAFR transcript 2 were increased by estrogen, but decreased by TGF-beta 1 in the human stomach cancer cell line (JR-St cells) which expressed both functional endogenous PAFR transcript 1 (leukocyte-type) and transcript 2 (tissue-type). Both ligands did not affect the expression of intrinsic PAFR transcript 1. Furthermore, the response elements to estrogen and TGF-beta 1 in the PAFR promoter 2 were delineated by a transient expression assay using the chloramphenicol acetyltransferase (CAT) gene as a reporter in this cell line. A negative response element for TGF-beta 1 was mapped on the sequence from -90 bp to -81 bp, which has consensus sequence for TIE (TGF-beta 1 inhibitory element). Although consensus estrogen response element (AGGTCAnnnTGACCT) is not present in this promoter, the entire sequence comprising two AGGTCA half motifs spaced by 153 bp (from -257 bp to -93 bp) conferred weak but significant estrogen responsiveness. Thus, through these elements in the PAFR promoter 2, estrogen and TGF-beta 1 may regulate the PAFR gene to achieve a tissue-specific expression. (C) 1994 Academic Press, Inc.
  • H MUTOH, S ISHII, T IZUMI, S KATO, T SHIMIZU
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 205 2 1137 - 1142 1994年12月 [査読有り][通常論文]
     
    The human platelet-activating factor receptor (PAFR) gene is transcribed by two distinct promoters (promoter 1 and promoter 2) to generate two transcripts (designated as PAFR transcript 1 and PAFR transcript 2), though their open reading frames are identical. By primer extension analysis to discriminate two transcripts, we found that the levels of PAFR transcript 1 (leukocyte-type), but not PAFR transcript 2 (tissue-type), are upregulated by PAF as well as by 12-O-tetradecanoylphorbol-13-acetate (TPA) in the human stomach cancer cell line (JR-ST cells) which expresses both functional PAFR transcript 1 and PAFR transcript 2 endogenously. Functional analysis of the promoter 1 with a transient expression assay using chloramphenicol acetyl-transferase (CAT) gene as a reporter showed that both PAF and TPA activated the promoter 1 but not the deleted promoter lacking the three consensus binding sites for NF-B-kappa located from -571 bp to -459 bp. These findings suggest a molecular mechanism of positive regulation of PAFR gene expression by PAF through NF-B-kappa, possibly by a phosphorylation reaction involving protein kinase C by PAF. (C) 1994 Academic Press, Inc.
  • K YOSHIURA, S OTA, A TERANO, M TAKAHASHI, Y HATA, T KAWABE, H MUTOH, H HIRAISHI, R NAKATA, K OKANO, M OMATA
    DIGESTIVE DISEASES AND SCIENCES 39 7 1454 - 1463 1994年07月 [査読有り][通常論文]
     
    We recently developed a primary culture system for gastric epithelial cells from adult rabbits that allows the investigation of growth regulation at the cellular level. In this study, we demonstrated that epidermal growth factor (EGF), insulin, and dibutyryl adenosine 3',5'-cyclic monophosphate (dBcAMP) all stimulated cell proliferation. Insulin and dBcAMP potentiated the stimulation of cell proliferation by EGF, while transforming growth factor-beta 1 (TGF-beta 1) inhibited it. Expression of c-fos and c-myc was induced in response to the stimulation by these growth regulators, but the degree of expression did not necessarily correlate with the effects of these agents on cell proliferation. In conclusion, EGF, insulin, and dBcAMP were positive growth regulators, while TGF-beta 1 was a negative regulator in gastric epithelial cells. These growth modulators may exert their effects by distinct pathways from a standpoint of the expression of c-fos and c-myc.
  • S OTA, K YOSHIURA, M TAKAHASHI, Y HATA, O KOHMOTO, T KAWABE, T SHIMADA, H HIRAISHI, H MUTOH, A TERANO, T SUGIMOTO, M OMATA
    GASTROENTEROLOGY 106 6 1485 - 1492 1994年06月 [査読有り][通常論文]
  • H HIRAISHI, A TERANO, S OTA, H MUTOH, T SUGIMOTO, T HARADA, M RAZANDI, KJ IVEY
    GASTROENTEROLOGY 106 5 1199 - 1207 1994年05月 [査読有り][通常論文]
  • M KURITA, Y NIWA, E HAMADA, Y HATA, M OSHIMA, H MUTOH, S SHIINA, R NAKATA, S OTA, A TERANO, T SUGIMOTO, M ONO, T SAWADA, M MORI, T NIKI, T OKA
    JOURNAL OF GASTROENTEROLOGY 29 2 208 - 213 1994年04月 [査読有り][通常論文]
     
    A case of Churg-Strauss syndrome with multiple perforations of the small intestine is described. A 31-year-old woman was admitted with a complaint of epigastric pain. She had a history of bronchial asthma. One week before admission, white blood cell count was 20 800/mm(3) with 59% eosinophils. Neurological examination on admission disclosed mononeuritis multiplex with paresthesia in both the lower and upper extremities. At colonoscopy, there were scattered aphthous ulcers in the colon. Ophthalmological examination revealed allergic conjunctivitis. After admission, hypereosinophilia increased to as high as 36 000/mm(3). Oral administration of prednisolone (60 mg/day) was begun. On the 3rd day of the treatment, the eosinophil count decreased dramatically, to 400/mm(3), while severe abdominal pain developed. Since abdominal X-ray him revealed free air in the abdominal cavity, emergency laparotomy was performed and multiple intestinal ulcers with perforations were found. Partial ileectomy was performed. Pathological findings of the resected specimen were interpreted as a necrotizing angiitis with extravascular granuloma. Since the operation, the patient has been asymptomatic, except for neurological symptoms. Hypereosinophilia has decreased without treatment to counts averaging 270/mm(3), within 3 months. On the basis of the clinical features and histopathological findings, a diagnosis of Churg-Strauss syndrome was established.
  • S OTA, Y HATA, A TERANO, K YOSHIURA, H HIRAISHI, T KAWABE, H MUTOH, S SHIINA, T SUGIMOTO
    DIGESTIVE DISEASES AND SCIENCES 38 8 1426 - 1434 1993年08月 [査読有り][通常論文]
     
    Prostaglandin (PG) has been reported to be one of the important protective factors in the gastric mucosa. However the mechanism of the regulation of endogenous PG production has not been well studied. We investigated the possible roles of Ca2+, cAMP, and protein kinase C (PKC) in the regulation of PGE2 release from cultured rabbit gastric mucosal cells. PGE2 was measured by radioimmunoassay. A23187 (Ca2+ ionophore) at 2 x 10(-6) M significantly increased PGE2 release. Deprivation of Ca2+ from the medium blocked the A23187-induced increase of PGE2. TMB-8 (a putative inhibitor of Ca2+ release from intracellular stores) did not have any significant effects on the increase of PGE2-induced by A23187. Thus, A23187 increased PGE2 through the influx of extracellular Ca2+. W7 or compound 48/80 (calmodulin inhibitors) did not alter the response of PGE2 caused by A23187. Exogenous administration of cAMP, forskolin (an activator of adenylate cyclase), or 2-chloroadenosine (a possible activator of adenylate cyclase through adenosine A2 receptor) had neither significant effects on PGE2 release nor an effect on A23187-induced increase of PGE2 release. 12-O-tetradecanoylphorbol 13-acetate (TPA, an activator of PKC) significantly stimulated PGE2 release in a dose-dependent fashion, whereas another phorbol ester with no biological activity did not. A23187 at 0.8 x 10(-6) M, but not cAMP, potentiated the TPA-induced increase of PGE2. Mepacrine (a phospholipase A2 inhibitor) reduced the A23187- and TPA-induced increase of PGE2. These results suggest that Ca2+ and protein kinase C may play important roles in the regulation of PGE2 release by cultured rabbit gastric cells.
  • Mutoh H, Bito H, Minami M, Nakamura M, Honda Z, Izumi T, Nakata R, Kurachi Y, Terano A, Shimizu T
    FEBS letters 322 2 129 - 134 2 1993年05月 [査読有り][通常論文]
  • H HIRAISHI, A TERANO, S OTA, H MUTOH, T SUGIMOTO, T HARADA, M RAZANDI, KJ IVEY
    JOURNAL OF LABORATORY AND CLINICAL MEDICINE 121 4 570 - 578 1993年04月 [査読有り][通常論文]
     
    The gastric epithelium is exposed to oxygen radicals that are generated within the lumen. Much interest has been focused on the role of mucus in maintaining integrity of the gastric mucosa against oxidants, because gastric mucus may act as a scavenger of oxygen radicals. The aim of this study was to assess the role of mucous glycoprotein in protecting cultured gastric epithelial cells against oxygen radicals. Monolayer cultures of rat gastric mucus-producing cells were studied. Oxygen radicals were generated by hypoxanthine and xanthine oxidase. Cytotoxicity was quantified by measuring chromium 51 release from prelabeled cells. Rate of mucous synthesis was estimated by incorporation of tritiated glucosamine into the cells. The effects of tetraprenyl acetone (a stimulant of mucus production) and N-acetyl-L-cysteine (a mucolytic agent) on oxygen radical-induced damage were determined. Preincubation with tetraprenyl acetone, while stimulating mucous glycoprotein by the cultured cells, caused a dose-dependent reduction of hypoxanthine-xanthine oxidase-induced Cr-51 release, reaching maximum protection of the damage by 31% to 50%. In contrast, pretreatment with N-acetyl-L-cysteine potentiated oxygen radical-induced Cr-51 release dose dependently. The protective effect of tetraprenyl acetone was significantly abolished by N-acetyl-L-cysteine. Neither tetraprenyl acetone nor N-acetyl-L-cysteine alone under the conditions of this study affected the cellular content of glutathione, which modulates oxygen radical injury to these cells. These results suggest that mucous glycoprotein partially but significantly protects cultured gastric epithelial cells against extracellularly generated oxygen radicals. It seems likely, therefore, that gastric mucus is involved in antioxidant defenses in these cells.
  • A TERANO, S OTA, H HIRAISHI, H MUTOH
    ACTA PATHOLOGICA JAPONICA 43 1-2 2 - 10 1993年01月 [査読有り][通常論文]
     
    In 1979, a new mechanism of gastric defense named cytoprotection was followed by numerous reports elucidating this interesting and important phenomenon. During this decade, however, the concept and definition of gastric cytoprotection have been modified from the morphological and ultrastructural viewpoints. This review attempts to describe the concept and mechanisms of cytoprotection as well as its pathophysiological features. Specifically, in vitro studies using isolated cells or monolayer cultured cells as well as molecular investigations of signal transduction system have been documented.
  • T SUGIMOTO, H TSUCHIMOCHI, CGA MCGREGOR, H MUTOH, T SHIMIZU, Y KURACHI
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 189 2 617 - 624 1992年12月 [査読有り][通常論文]
  • Sakanaka C, Izumi T, Nakamura M, Honda Z, Watanabe T, Minami M, Mutoh H, Bito H, Seyama Y, Ui M
    Biochimica et biophysica acta 1175 61 - 66 1 1992年12月 [査読有り][通常論文]
  • M MINAMI, H BITO, N OHISHI, H TSUGE, M MIYANO, M MORI, H WADA, H MUTOH, S SHIMADA, T IZUMI, K ABE, T SHIMIZU
    FEBS LETTERS 309 3 353 - 357 1992年09月 [査読有り][通常論文]
     
    We previously obtained evidence for intrinsic aminopeptidase activity for leukotriene (LT)A4 hydrolase. an enzyme characterized to specifically catalyse the hydrolysis of LTA4 to LTB4, a chemotactic compound. From a sequence homology search between LTA4 hydrolase and several aminopeptidases, it became clear that they share a putative active site for known aminopeptidases and a zinc binding domain. Thus, Glu-297 of LTA4 hydrolase is a candidate for the active site of its aminopeptidase activity, while His-296, His-300 and Glu-319 appear to constitute a zinc binding site. To determine whether or not this putative active site is also essential to LTA4 hydrolase activity, site-directed mutagenesis experiments were carried out. Glu-297 was mutated into 4 different amino acids. The mutant E-297Q (Glu changed to Gln) conserved LTA4 hydrolase activity but showed little aminopeptidase activity. Other mutants at Glu-297 (E297A, E297D and E297K) showed markedly reduced amounts of both activities. It is thus proposed that either a glutamic or glutamine moiety at 297 is required for full LTA4 hydrolase activity, while the free carboxylic acid of glutamic acid is essential for aminopeptidase.
  • Ota S, Hata Y, Hiraishi H, Mutoh H, Terano A, Sugimoto T
    Journal of clinical gastroenterology 14 Suppl 1 S156 - 61 1992年 [査読有り][通常論文]
  • Hiraishi H, Terano A, Ota S, Mutoh H, Sugimoto T, Razandi M, Ivey KJ
    The American journal of physiology 261 G921 - 8 6 Pt 1 1991年12月 [査読有り][通常論文]
  • Hiraishi H, Terano A, Ota S, Mutoh H, Sugimoto T, Razandi M, Ivey KJ
    The American journal of physiology 261 G662 - 8 4 Pt 1 1991年10月 [査読有り][通常論文]
  • M NAKAMURA, Z HONDA, T IZUMI, C SAKANAKA, H MUTOH, M MINAMI, H BITO, Y SEYAMA, T MATSUMOTO, M NOMA, T SHIMIZU
    JOURNAL OF BIOLOGICAL CHEMISTRY 266 30 20400 - 20405 1991年10月 [査読有り][通常論文]
     
    The cDNA for a platelet-activating factor (PAF) receptor was cloned from a human leukocyte cDNA library using a 0.8-kilobase pair fragment of the guinea pig lung PAF receptor cDNA (Honda, Z., Nakamura, M., Miki, I., Minami, M., Watanabe, T., Seyama, Y., Okado, H., Toh, H., Ito, K., Miyamoto, T., and Shimizu, T. (1991) Nature 349, 342-346). The cDNA (1.8-kilobase pairs) had an open reading frame encoding 342 amino acid residues with a calculated M(r) of 39,203. The clone was shown to code for a PAF receptor based on the following criteria: 1) the amino acid sequence possesses seven putative membrane spanning domains with 83% identity to the guinea pig lung PAF receptor, 2) Xenopus laevis oocytes injected with the transcript of the clone showed an electrophysiological response to PAF, and 3) COS-7 cells expressing the encoded receptor showed ligand binding with the pharmacological properties of the PAF receptor. Activation of the PAF receptor yielded inositol 1,4,5-trisphosphate production in both COS-7 cells and oocytes, and guanosine 5'-O-(2-thio)bisphosphate injection into the oocytes inhibited PAF-induced Cl- current, providing an evidence that PAF stimulates phosphoinositide turnover via G-protein(s). PAF receptor mRNA was abundant in leukocytes and less so in an undifferentiated human eosinophilic cell line (EoL-1 cells) or human erythroleukemia cells (HEL cells). The production of the mRNA was prominently increased when EoL-1 cells were treated with granulocyte macrophage colony stimulating factor, interleukin-5, and n-butyrate.
  • S OTA, H HIRAISHI, A TERANO, H MUTOH, Y HATA, KJ IVEY, T SUGIMOTO
    JAPANESE JOURNAL OF PHARMACOLOGY 57 1 63 - 69 1991年09月 [査読有り][通常論文]
     
    While cimetidine (CIM) is strikingly effective in inhibiting gastric acid secretion, its effect on the defensive mechanisms of the gastric mucosa has been controversial. The aims of the present study were to test if administration of CIM at an antisecretory dose is protective against acid-induced injury and to assess its effect on adaptive cytoprotection induced by non-necrotizing concentrations of HCl in rats. A dose of 100 mg/kg of CIM was administered once, or twice a day for 5 days intraperitoneally. To study the effect of CIM on HCl-induced damage, 0.6 N HCl was given orally one hour after the last administration of CIM. To study the effect of CIM on adaptive cytoprotection, 0.35 N HCl was given orally one hour after the last administration of CIM. Fifteen minutes later, 0.6 N HCl was given orally. Thirty minutes after the administration of 0.6 N HCl, the stomach was removed and ulcer indices were calculated. Pretreatment with CIM did not prevent 0.6 N HCl induced gastric damage. Prior administration of 0.35 N HCl significantly reduced ulcer indices caused by 0.6 N HCl. Short or long term treatment with CIM did not have significant effects on the reduction of ulcer indices. These results suggest that CIM at an antisecretory dose neither acts as a protective agent nor modulates the protective process of the gastric mucosa.
  • Mutoh H, Ota S, Hiraishi H, Ivey KJ, Terano A, Sugimoto T
    The American journal of physiology 261 G65 - 70 1 Pt 1 1991年07月 [査読有り][通常論文]
  • Hiraishi H, Terano A, Ota S, Mutoh H, Razandi M, Sugimoto T, Ivey KJ
    The American journal of physiology 260 G556 - 63 4 Pt 1 1991年04月 [査読有り][通常論文]
  • S OTA, H TSUKAHARA, A TERANO, Y HATA, H HIRAISHI, H MUTOH, T SUGIMOTO
    DIGESTIVE DISEASES AND SCIENCES 36 4 409 - 416 1991年04月 [査読有り][通常論文]
     
    Ursodeoxycholate (UDC) and tauroursodeoxycholate (TUDC) have been reported to be protective against liver injury induced by other bile salts. UDC also has been shown to be effective against refluxed bile-induced gastritis after gastric surgery. However the mechanism of the therapeutic effect of UDC on gastric mucosa has not been known. In the present study, cytoprotective actions of UDC and TUDC against chenodeoxycholate (CDC) -induced gastric injury were investigated using rabbit gastric cell cultures without systemic factors. Rabbit gastric mucosal cells were cultured after the isolation of rabbit gastric cells with collagenase and ethylenediaminetetraacetic acid. Cytotoxicity was quantified by measuring Cr-51 release from prelabeled cells and MTT assay. Prostaglandin (PG) E2 was assayed by radioimmunoassay. Concentrations of CDC > 0.5mM or UDC > 5mM caused cellular damage and increased Cr-51 release in a dose-dependent and time-dependent fashion, while TUDC up to 10 mM did not. TUDC, but not UDC, showed a significant decrease of CDC (1.5 mM) -induced Cr-51 release dose dependently. The protective effect of TUDC against CDC-induced damage was confirmed by MTT assay. On phase-contrast microscopy, disruption of monolayers induced by CDC (1.5 mM) was clearly protected by TUDC (10 mM). Free radical scavengers (500 units/ml of superoxide dismutase, 300 units/ml of catalase, and 100 mM of dimethyl sulfoxide) or a calcium blocker (10(-7)-10(-5) M verapamil) did not show significant protection against CDC-induced damage. Deprivation of Ca2+ in the media did not affect CDC-induced damage. Thus free radicals or Ca2+ might not be involved in the cell toxicity of CDC. Although TUDC (10 mM) significantly increased PGE2 production by cultured cells, indomethacin (10(-4) M) did not reduce protective effects of TUDC, as assessed by Cr-51 release and MTT assay. In conclusion TUDC is cytoprotective against CDC-induced damage to cultured rabbit gastric cells. Neither free radicals, Ca2+, nor endogenous PGs may play leading roles in the mechanism of this action.
  • Shuichiro Shiina, Yasuo Hata, Yasuro Niwa, Yutaka Komatsu, Torao Tanaka, Kenta Yoshiura, Eiji Hamada, Masamichi Ohshima, Hiroyuki Mutoh, Masahiro Kurita, Ryo Nakata, Shinichi Ota, Yasushi Shiratori, Akira Terano, Tsuneaki Sugimoto, Makoto Taniguchi, Yoshiki Uta, Hiroaki Tsukahara, Kazumi Tagawa, Tadao Unuma, Takao Kawabe, Ken'ichi Okano
    Gastroenterologia Japonica 26 1 47 - 50 1991年02月 [査読有り][通常論文]
     
    One of the shortcomings of percutaneous ethanol injection therapy (PEIT) is that many sessions are necessary to accomplish the treatment. In order to reduce the number of treatment sessions, we inserted two or three needles before injection of ethanol was begun. Using the multiple-needle insertion method, we markedly reduced the number of treatment sessions. Histopathologic examination, imaging techniques, and serum alpha-fetoprotein levels showed efficacy of PEIT using the multiple-needle insertion method. No serious complication occurred. Levels of transient pain, fever, and the feeling of intoxication did not seem to be different from those occurring with the conventional method. Multiple-needle insertion method may be valuable as a method for reducing the number of treatment sessions necessary and thus shortening the treatment period. © 1991 The Japanese Society of Gastroenterology.
  • Akira Terano, Ryo Nakada, Hiroyuki Mutoh, Hideyuki Hiraishi, Shinichi Ota, Shuichiro Shiina, Tadato Shimada, Yasuaki Itoh, Kenjiro Kimura, Junji Shiga, Tsuneaki Sugimoto
    Gastroenterologia Japonica 26 1 7 - 13 1991年02月 [査読有り][通常論文]
     
    Despite the importance of in vitro study of gastric cancer, there are very few established cell lines derived from human gastric carcinoma. We have recently established a new cell line derived from human gastric cancer which has the ability to produce tumor markers. This cell line has been designated JR-St. This cell line was derived from the cerebrospinal fluid of a 37-yr-old female patient who had metastatic brain tumor of signet ring cell gastric adenocarcinoma. This cell line has been maintained for more than 24 months through 80 passages with stable growth. PAS staining showed intracellular mucin granules. Transmission and scanning electron microscopy revealed cells with numerous microvilli and fine projections as well as intracellular granules, indicating mucin. This cell line had the ability to produce high concentrations of tumor markers such as carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 19-9. Thus this cell line should provide a very useful tool for the investigation of gastric cancer such as analysis of tumor markers as well as effects of anti-cancer drugs or growth factors. © 1991 The Japnnese Society of Gastroenterology.
  • A TERANO, J SHIGA, H MUTOH, H HIRAISHI, S SHIINA, M KURITA, S OTA, Y ITOH, T SUGIMOTO
    JAPANESE JOURNAL OF PHARMACOLOGY 55 1 115 - 120 1991年01月 [査読有り][通常論文]
     
    Tetraprenylacetone (TPA: teprenon, geranylgeranylacetone) is a novel anti-ulcer agent developed in Japan. The aim of this study was to test whether TPA has the ability to promote the healing process of rat gastric mucosal injury induced by absolute ethanol (ET). Fasted rats received orally 5 ml/kg of absolute ET. Sixty minutes later, TPA (200 mg/kg) or saline (control) was administered intragastrically. Thereafter, the same dose of TPA or saline was given orally every 8 hours. To investigate the role of endogenous prostaglandins, indomethacin was given intraperitoneally every 8 hours. Twenty four or 48 hours after the first administration of TPA or saline, rats were sacrificed and the stomachs were removed. Administration of TPA significantly reduced lesion indices from 100 +/- 12.9% (control) to 57.0 +/- 12.8% (24 hours, P < 0.05) and from 100 +/- 15.3% (control) to 17.6 +/- 3.4% (48 hours, P < 0.01). Addition of indomethacin did not significantly affect this effect of TPA. Ultrastructual studies revealed that TPA stimulated regeneration of gastric mucosa damaged by ET after 24 and 48 hours. These results indicate that TPA has the ability to promote the healing process of gastric mucosal damage induced by absolute ET. It is, however, unlikely that endogenous prostaglandins are involved in this promotive effect of TPA on the healing process of gastric injury.
  • M MINAMI, N OHISHI, H MUTOH, T IZUMI, H BITO, H WADA, Y SEYAMA, H TOH, T SHIMIZU
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 173 2 620 - 626 1990年12月 [査読有り][通常論文]
  • S SHIINA, H AOYAMA, Y SHIRATORI, H MUTOH, M KURITA, S OTA, A TERANO, T SUGIMOTO
    INVESTIGATIVE RADIOLOGY 25 6 651 - 657 1990年06月 [査読有り][通常論文]
  • H MUTOH, H HIRAISHI, S OTA, H YOSHIDA, KJ IVEY, A TERANO, T SUGIMOTO
    GASTROENTEROLOGY 98 6 1452 - 1459 1990年06月 [査読有り][通常論文]
  • Mutoh H, Hiraishi H, Ota S, Ivey KJ, Terano A, Sugimoto T
    The American journal of physiology 258 G603 - 9 4 Pt 1 1990年04月 [査読有り][通常論文]
  • S OTA, H HIRAISHI, A TERANO, H MUTOH, Y KURACHI, T SHIMADA, KJ IVEY, T SUGIMOTO
    DIGESTIVE DISEASES AND SCIENCES 34 12 1882 - 1889 1989年12月 [査読有り][通常論文]
  • Shuichiro Shiina, Hiromu Aoyama, Yasushi Shiratori, Yasuo Hata, Yasuo Niwa, Yutaka Komatsu, Hiroyuki Mutoh, Masahiro Kurita, Ryo Nakata, Tadahito Shimada, Takao Kawabe, Tateo Kawase, Shinichi Ota, Akira Terano, Tsuneaki Sugimoto
    Gastroenterologia Japonica 24 6 740  1989年12月 [査読有り][通常論文]
  • Terano A, Nakada R, Mutoh H, Hiraishi H, Ota S, Kimura K, Aoyama H, Sugimoto T, Shiga J, Itoh Y
    Human cell : official journal of Human Cell Research Society 2 307 - 309 3 1989年09月 [査読有り][通常論文]
  • A. Terano, R. Nakada, H. Hiraishi, S. Ota, H. Mutoh, T. Shimada, S. Shiina, Y. Itoh, J. Shiga, T. Sugimoto
    Gastroenterologia Japonica 24 2 219  1989年04月 [査読有り][通常論文]
  • A. Terano, J. Shiga, S. Ota, H. Hiraishi, H. Mutoh, T. Sugimoto
    Gastroenterologia Japonica 24 1 81  1989年02月 [査読有り][通常論文]
  • A. Terano, J. Shiga, H. Hiraishi, S. Ota, H. Mutdh, T. Sugimoto
    Gastroenterologia Japonica 24 1 80  1989年02月 [査読有り][通常論文]

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