研究者総覧

鈴木 浩一 (スズキ コウイチ)

  • 総合医学第2講座 准教授
メールアドレス: ksuzbnhmyahoo.co.jp
Last Updated :2020/07/29

研究者情報

学位

  • 医学博士(1999年03月 浜松医科大学)

学位

    鈴木 浩一

論文上での記載著者名

  • Koichi Suzuki
  • Suzuki K

科研費研究者番号

  • 70332369

J-Global ID

研究分野

  • ライフサイエンス / 消化器外科学

研究活動情報

論文

  • Keiko Akahane, Katsuyuki Shirai, Masaru Wakatsuki, Kazunari Ogawa, Kyosuke Minato, Kohei Hamamoto, Satoru Takahashi, Koichi Suzuki, Jun Takahashi, Toshiki Rikiyama, Keita Matsumoto, Hirosato Mashima
    Clinical case reports 8 5 919 - 922 2020年05月 [査読有り][通常論文]
     
    Antiangiogenic agents, such as ramucirumab, should be cautiously administered along with radiotherapy because of the enhanced risk of adverse events.
  • Taro Fukui, Koichi Suzuki, Sawako Tamaki, Iku Abe, Yuhei Endo, Hideki Ishikawa, Nao Kakizawa, Fumiaki Watanabe, Masaaki Saito, Shingo Tsujinaka, Kazushige Futsuhara, Yasuyuki Miyakura, Hiroshi Noda, Toshiki Rikiyama
    Surgical case reports 5 1 145 - 145 2019年10月 [査読有り][通常論文]
     
    BACKGROUND: Anti-epidermal growth factor receptor (EGFR) antibody is widely used for the treatment of patients with metastatic colorectal cancer. Hypomagnesemia is a comparatively frequent adverse event of this drug, which is likely overlooked because it occurs later in treatment without symptoms. Furthermore, hypomagnesemia and hypomagnesemia-induced corrected QT (QTc) prolongation may lead to loss of consciousness (LOC), the onset of which is not generally considered associated with the treatment of anti-EGFR antibody because of its rare occurrence. Here, we present a colorectal cancer patient treated with anti-EGFR antibody, who suffered LOC during treatment while severe hypomagnesemia or QTc prolongation was not observed. CASE PRESENTATION: A 69-year-old man with metastatic colon cancer was treated with cetuximab (anti-EGFR antibody) plus irinotecan as third-line chemotherapy. His serum magnesium level gradually decreased, and grade 2 hypomagnesemia (a serum magnesium level of 0.9 mg/dL) was observed at the 12th administration of cetuximab. In light of this development, intravenous supplementation of 20 mEq magnesium sulfate began with careful blood monitoring despite the lack of clinical symptoms. Electrocardiogram (ECG) showed prolonged QT or corrected QT (QTc) intervals (grade 1). His serum magnesium level remained at 0.9 mg/dL, and no hypomagnesemia symptoms were observed by the 17th administration of cetuximab. After the treatment, however, he suddenly lost consciousness without symptoms related to infusion or allergic reactions. Circulatory collapse following dermatological reactions and respiratory events were not evident. Intravenous supplementation of magnesium sulfate was administered again. He awakened 2 min after the onset of temporary LOC without any other symptoms related to hypomagnesemia, such as lethargy, tremor, tetany, and seizures. No other etiology outside of the low level of serum magnesium was confirmed in further examinations. Cetuximab was discontinued, and his serum magnesium level returned to a level within the normal range after 6 weeks. Because of tumor progression, regorafenib and TAS-102 (trifluridine tipiracil hydrochloride) were introduced sequentially for 6 months. Five months after the final treatment of TAS-102, he died of his primary disease, which reflected a survival period of 4 years and 6 months since the beginning of treatment. CONCLUSIONS: This case report reminds clinicians that LOC can be induced without severe hypomagnesemia or QTc prolongation, during anti-EGFR antibody treatment for metastatic colorectal cancer even while under carefully monitored magnesium supplementation.
  • Nao Kakizawa, Koichi Suzuki, Iku Abe, Yuhei Endo, Sawako Tamaki, Hideki Ishikawa, Fumiaki Watanabe, Kosuke Ichida, Masaaki Saito, Kazusige Futsuhara, Fumio Konishi, Toshiki Rikiyama
    Oncology reports 42 2 857 - 865 2019年08月 [査読有り][通常論文]
     
    Patients with breast cancer who undergo surgery have a risk of developing multiple cancers in the contralateral breast and other organs. We previously reported that overexpression of satellite alpha transcripts (SAT) facilitates chromosomal instability, which is involved in the development of multiple tumors in patients with colorectal and gastric cancer. In this study, we elucidated the significance of SAT in the development of multiple tumors in patients with breast cancer. Relative expression of SAT (rSAT) was calculated in normal and tumor tissues from 167 patients. In total, 27 patients developed bilateral breast cancer (BBC) and 27 patients showed multiple primary cancer (MPC), with patients with BBC and MPC showing higher rSAT levels in tumor tissues than those in patients with single breast cancer (SBC) (P=0.0312 and P=0.0420, respectively). Additionally, higher rSAT levels in tumor tissues from patients with BBC were a significant factor according to univariate analysis, and multivariate analysis showed that rSAT >1.5 was a significant predictor of MPC [hazard ratio (HR): 2.96; P=0.0243); however, we did not clarify the involvement of SAT in normal tissues. Excluding 71 patients with BRCA‑related clinical features, rSAT levels were higher in patients with BBC and MPC than in patients with SBC in tumor tissues and normal tissues (P<0.05). Significant predictors according to univariate analysis included rSAT >1.5 in tumor tissues, rSAT >2.4 in normal tissues, and T <2, whereas those for multivariate analysis included rSAT >2.4 in normal tissues for BBC (HR: 22.7; P=0.00120) and MPC (HR: 13.0; P=0.00601). Our data indicated that patients with breast cancer and high rSAT levels in their breast tissues exhibit a 10‑ to 20‑fold increased risk for the development of multiple cancers when harboring no BRCA‑related clinical features.
  • Yuta Muto, Koichi Suzuki, Takaharu Kato, Kosuke Ichida, Yuji Takayama, Taro Fukui, Nao Kakizawa, Fumiaki Watanabe, Yuji Kaneda, Hiroshi Noda, Toshiki Rikiyama
    Molecular and clinical oncology 10 5 511 - 515 2019年05月 [査読有り][通常論文]
     
    As a result of recent advances in diagnostic techniques and treatment modalities, the number of patients diagnosed with multiple primary malignancies has been increasing. We report the case of a 79-year-old male with multiple primary malignancies of three histological types in six different organs: Stomach, prostate, colon, urinary bladder, facial skin and pancreas, in chronological order. The first malignancy was upper gastric cancer diagnosed in 1998. The second and third malignancies were prostate cancer and ascending colon cancer, which were diagnosed in 2010. The fourth malignancy was bladder cancer diagnosed in 2011. The fifth and sixth malignancies were squamous cell skin cancer of the right cheek and intraductal papillary mucinous carcinoma (IPMC), respectively, diagnosed in 2014. The gastric cancer, colon cancer, bladder cancer, skin cancer and IPMC were surgically resected. The prostate cancer was treated by anti-androgen therapy. The patient died of local recurrence of IPMC in August 2016. Although multiple primary malignancies are not uncommon, diagnosis of six primary malignancies in a single patient, as reported in the present study, is extremely rare. It is important to understand the characteristics of multiple primary malignancies in order to administer suitable treatment and determine relevant follow-up plans for patients with cancer.
  • Tsutomu Takenami, Shingo Tsujinaka, Jun Takahashi, Sawako Tamaki, Ryo Maemoto, Rintaro Fukuda, Hideki Ishikawa, Nao Kakizawa, Fumi Hasegawa, Rina Kikugawa, Yasuyuki Miyakura, Koichi Suzuki, Akira Tanaka, Toshiki Rikiyama
    Case reports in surgery 2019 8129358 - 8129358 2019年 [査読有り][通常論文]
     
    Introduction: We herein present three cases of locally advanced colon cancer (LACC) invading the urinary bladder, in whom combined neoadjuvant chemotherapy with surgical intervention was effective in disease control and preserving urinary function. Case Presentation: Before neoadjuvant chemotherapy, all three cases underwent loop transverse colostomy for symptomatic colonic obstruction. Case 1: after 6 courses of capecitabine plus oxaliplatin (CAPOX), we performed sigmoid colectomy and partial resection of the bladder. The histological examination revealed pathological complete response (pCR). The final diagnosis was ypStage 0 (ypT0ypN0M0). Case 2: after 13 courses of CAPOX plus bevacizumab, we performed Hartmann's operation with partial resection of the bladder. The histological examination revealed pCR. The final diagnosis was ypStage 0 (ypT0ypN0M0). Case 3: after 6 courses of chemotherapy with CAPOX plus bevacizumab, we performed sigmoid colectomy and partial resection of the bladder. The pathological response was grade 1a according to the Japanese Classification of Colorectal Carcinoma. The final diagnosis was ypStage IIC (ypT4bypN0M0). All three cases underwent capecitabine-based adjuvant chemotherapy after radical surgery and patients are alive without recurrence. Conclusion: Neoadjuvant chemotherapy with CAPOX with or without bevacizumab followed by radical surgery could be an effective treatment option for LACC invading the urinary bladder.
  • Fumiaki Watanabe, Koichi Suzuki, Sawako Tamaki, Iku Abe, Yuhei Endo, Yuji Takayama, Hideki Ishikawa, Nao Kakizawa, Masaaki Saito, Kazushige Futsuhara, Hiroshi Noda, Fumio Konishi, Toshiki Rikiyama
    PloS one 14 12 e0227366  2019年 [査読有り][通常論文]
     
    BACKGROUND: Liquid biopsies enable the detection of circulating tumor DNA (ctDNA). However, the clinical significance of KRAS-mutated ctDNA for pancreatic cancer has been inconsistent with respect to its prognostic and predictive potential. METHODS AND FINDINGS: A total of 422 blood samples were collected from 78 patients undergoing treatments for localized and metastatic pancreatic ductal adenocarcinoma. KRAS mutation in tissues and KRAS ctDNA levels in plasma were determined by RASKET and droplet digital polymerase chain reaction. Longitudinal monitoring of KRAS ctDNA was performed to assess its significance for predicting recurrence and prognosis and for evaluating therapeutic responses to chemotherapy compared with carbohydrate antigen 19-9 (CA19-9). In 67 tumor tissues, discrepancies in point mutations of KRAS were rarely observed among individual patients, implying that one targeted point mutation of KRAS can be determined in tumor tissues prior to longitudinal blood monitoring. One-time blood assessment of KRAS-mutated ctDNA before surgery or chemotherapy was not clearly associated with recurrence and prognosis. Sequential blood monitoring was performed in 39 patients who underwent surgery for potentially resectable tumors. Increased CA19-9 levels were significantly associated with recurrence, but not prognosis (P<0.001, P = 1.0, respectively), whereas emergence of KRAS ctDNA was significantly associated with prognosis (P<0.001) regardless of recurrence. Furthermore, in 39 patients who did not undergo surgery, detection of KRAS ctDNA was a predictive factor for prognosis (P = 0.005). Multivariate analysis revealed that detection of KRAS ctDNA was the only independent prognostic factor regardless of tumor resection (hazard ratios = 54.5 for patients who underwent surgery and 10.1 for patients who did not undergo surgery; P<0.001 for both). Patients without emergence of KRAS ctDNA within 1 year after surgery showed significantly better prognosis irrespective of recurrence (P<0.001). No detection or disappearance of KRAS ctDNA within 6 months of treatment was significantly correlated with therapeutic responses to first-line chemotherapy (P<0.001). Changes in KRAS status provided critical information for the prediction of therapeutic responses. CONCLUSIONS: Our study showed for the first time that detection of KRAS ctDNA levels within a short period enables the prediction of prognosis and therapeutic responses in patients with pancreatic cancer.
  • Yuji Takayama, Koichi Suzuki, Yuta Muto, Kosuke Ichida, Taro Fukui, Nao Kakizawa, Hideki Ishikawa, Fumiaki Watanabe, Fumi Hasegawa, Masaaki Saito, Shingo Tsujinaka, Kazushige Futsuhara, Yasuyuki Miyakura, Hiroshi Noda, Fumio Konishi, Toshiki Rikiyama
    Oncotarget 9 36 24398 - 24413 2018年05月 [査読有り][通常論文]
     
    KRAS mutated circulating tumor DNA (MctDNA) can be monitored in the blood of patients with metastatic colorectal cancer (mCRC), but dynamic changes have not been determined. Four hundred and fifty-seven plasma samples were collected prospectively from 85 mCRC patients who underwent chemotherapy. MctDNA in plasma was detected by droplet digital PCR, and the percentage of MctDNA in total circulating cell-free DNA was calculated. KRAS assessment in tumor tissues showed 29 patients with the mutant-type (MT) and 56 patients with the wild-type (WT). Twenty-three of 29 MT patients (79.3%) and 28 of 56 WT patients (50.0%) showed MctDNA. Emergence of MctDNA was recognized during treatments with various drugs. Regardless of KRAS status in tumor tissues, patients with MctDNA in blood showed poor progression-free survival with first-line treatment. Median percentage of MctDNA accounted for 10.10% in MT patients and 0.22% in WT patients. These differences between MT and WT likely affected patterns of changes in MctDNA. KRAS monitoring identified dynamic changes in MctDNA, such as continuous, intermittent, and transient changes (quick elevation and disappearance). Emergence of MctDNA involved drug resistance, except for transient changes, which were seen in WT patients and likely corresponded with the drug response. Transient changes could be involved in recovery of sensitivity to anti-EGFR antibody in WT patients. Monitoring MctDNA during various treatments showed dynamic changes in KRAS status and could provide useful information for determining treatments for patients with mCRC.
  • Kosuke Ichida, Koichi Suzuki, Taro Fukui, Yuji Takayama, Nao Kakizawa, Fumiaki Watanabe, Hideki Ishikawa, Yuta Muto, Takaharu Kato, Masaaki Saito, Kazushige Futsuhara, Yasuyuki Miyakura, Hiroshi Noda, Tsukasa Ohmori, Fumio Konishi, Toshiki Rikiyama
    International journal of oncology 52 5 1685 - 1693 2018年05月 [査読有り][通常論文]
     
    The impairment of the stability of the chromosomal structure facilitates the abnormal segregation of chromosomes, thus increasing the risk of carcinogenesis. Chromosomal stability during segregation is managed by appropriate methylation at the centromere of chromosomes. Insufficient methylation, or hypomethylation, results in chromosomal instability. The centromere consists of satellite alpha repetitive sequences, which are ideal targets for DNA hypomethylation, resulting in the overexpression of satellite alpha transcript (SAT). The overexpression of SAT has been reported to induce the abnormal segregation of chromosomes. In this study, we verified the oncogenic pathway via chromosomal instability involving DNA hypomethylation and the overexpression of SAT. For this purpose, we constructed lentiviral vectors expressing SAT and control viruses and then infected human mammary epithelial cells with these vectors. The copy number alterations and segregation errors of chromosomes were evaluated by microarray-based comparative genomic hybridization (array CGH) and immunocytochemistry, respectively. The levels of hypomethylation of satellite alpha sequences were determined by MethyLight polymerase chain reaction. Clinical specimens from 45 patients with breast cancer were recruited to verify the data in vitro. The results of immunocytochemistry revealed that the incidence of segregation errors was significantly higher in the cells overexpressing SAT than in the controls. An array CGH identified the specific chromosomes of 8q and 20q as frequent sites of copy number alterations in cells with SAT overexpression, although no such sites were noted in the controls, which was consistent with the data from clinical specimens. A regression analysis revealed that the expression of SAT was significantly associated with the levels of hypomethylation of satellite alpha sequences. On the whole, the overexpression of SAT led to chromosomal instability via segregation errors at specific chromosomes in connection with DNA hypomethylation, which was also recognized in clinical specimens of patients with breast cancer. Thus, this oncogenic pathway may be involved in the development of breast cancer.
  • Nao Kakizawa, Hiroshi Noda, Fumiaki Watanabe, Kosuke Ichida, Koichi Suzuki, Toshiki Rikiyama
    World journal of surgery 42 4 1129 - 1137 2018年04月 [査読有り][通常論文]
     
    BACKGROUND: To evaluate the clinical significance of a CT-based evaluation of abdominal aortic calcification (AAC) in the postoperative outcomes after pancreaticoduodenectomy (PD) in elderly patients. METHODS: Patients 70 years of age and older who were randomly assigned to Group A were compared with those younger than 70 who were assigned to Group B in terms of preoperative and intraoperative variables and postoperative outcomes. We compared the patients with clinically relevant postoperative pancreatic fistula (CR-POPF) (Group C) to those without CR-POPF (Group D), and especially Group A. We also compared the patients with CR-POPF (Group E) to those without CR-POPF (Group E) to clarify the risk factors for POPF, in each of the analyses. The AAC score was determined using the methods of Agatston et al. RESULTS: Group A more often had frequent atherosclerosis-related comorbidities (62.2%), low serum albumin (55.9%), and a high AAC score (66.1%). There were no significant differences in the postoperative variables. The comparisons between Groups C and D identified four independent risk factors for CR-POPF: BMI ≥ 25 (OR 8.54, 95% CI 3.15-23.1), male gender (OR 3.17, 95% CI 1.28-7.85), soft pancreatic parenchyma (OR 3.43, 95% CI 1.34-8.81), and the absence of MPD dilatation (OR 5.70, 95% CI 2.13-15.3). Comparisons between Groups E and F identified two independent risk factors for CR-POPF: BMI ≥ 25 (OR 29.4, 95% CI 5.77-150) and a high ACC score (OR 10.8, 95% CI 2.08-56.6). CONCLUSIONS: We demonstrated, for the first time, that a high AAC score is a risk factor of CR-POPF in elderly patients who underwent PD.
  • Sergio Alonso, Koichi Suzuki, Fumiichiro Yamamoto, Manuel Perucho
    Methods in molecular biology (Clifton, N.J.) 1766 137 - 156 2018年 [査読有り][通常論文]
     
    Somatic, and in a minor scale also germ line, epigenetic aberrations are fundamental to carcinogenesis, cancer progression, and tumor phenotype. DNA methylation is the most extensively studied and arguably the best understood epigenetic mechanisms that become altered in cancer. Both somatic loss of methylation (hypomethylation) and gain of methylation (hypermethylation) are found in the genome of malignant cells. In general, the cancer cell epigenome is globally hypomethylated, while some regions-typically gene-associated CpG islands-become hypermethylated. Given the profound impact that DNA methylation exerts on the transcriptional profile and genomic stability of cancer cells, its characterization is essential to fully understand the complexity of cancer biology, improve tumor classification, and ultimately advance cancer patient management and treatment. A plethora of methods have been devised to analyze and quantify DNA methylation alterations. Several of the early-developed methods relied on the use of methylation-sensitive restriction enzymes, whose activity depends on the methylation status of their recognition sequences. Among these techniques, methylation-sensitive amplification length polymorphism (MS-AFLP) was developed in the early 2000s, and successfully adapted from its original gel electrophoresis fingerprinting format to a microarray format that notably increased its throughput and allowed the quantification of the methylation changes. This array-based platform interrogates over 9500 independent loci putatively amplified by the MS-AFLP technique, corresponding to the NotI sites mapped throughout the human genome.
  • Jun Takahashi, Shingo Tsujinaka, Nao Kakizawa, Noriya Takayama, Erika Machida, Kazuki Iseya, Fumi Hasegawa, Rina Kikugawa, Yasuyuki Miyakura, Koichi Suzuki, Toshiki Rikiyama
    Case reports in surgery 2018 1674279 - 1674279 2018年 [査読有り][通常論文]
     
    Recent advancements in multimodal therapy can provide oncologic benefits for patients with recurrent colorectal cancer. This report presents a case of locoregionally recurrent appendiceal cancer treated with neoadjuvant chemotherapy followed by surgical resection with vascular reconstruction. A 68-year-old Japanese woman was diagnosed with appendiceal cancer and underwent ileocecal resection. The pathological evaluation revealed KRAS-mutant adenocarcinoma with the final stage of T4bN1M0. She received oral fluorouracil-based adjuvant chemotherapy. One year later, she was found to have peritoneal dissemination in the pelvic cavity and vaginal metastasis. She received an oxaliplatin-based chemotherapy followed by surgical resection. One year after the second surgery, she developed a locoregional recurrence involving the right external iliac vessels and small intestine. She received an irinotecan-based regimen with bevacizumab as neoadjuvant chemotherapy, followed by surgical resection. At first, a femoro-femoral bypass was made to secure the blood supply to the right lower extremities. Subsequently, an en bloc resection including the recurrent tumor and the external iliac vessels was completed. Surgical resection for recurrent colorectal cancer is often technically challenging because of the tumor location and invasion to adjacent organs. In this case, a surgical approach with persistent chemotherapy achieved oncologic resection of locoregionally recurrent appendiceal cancer.
  • Kazuhisa Hosoya, Satoshi Matsusaka, Tomomi Kashiwada, Koichi Suzuki, Norio Ureshino, Akemi Sato, Yoshio Miki, Kazuki Kitera, Mitsuharu Hirai, Kiyohiko Hatake, Shinya Kimura, Naoko Sueoka-Aragane
    Pathology oncology research : POR 23 4 737 - 744 2017年10月 [査読有り][通常論文]
     
    KRAS mutations have been recognized as predictive markers of primary resistance to anti-EGFR-antibodies in colorectal cancer patients. In addition, newly detected KRAS mutations have been reported to be related with acquired resistance to chemotherapy containing anti-EGFR antibody. Considering this evidence, monitoring of KRAS mutations is indispensable for making treatment decisions, and the method should be non-invasive allowing repeated examinations. Recently, we established a novel automated sensitive detection system for KRAS mutations, named mutation-biased PCR quenching probe system (MBP-QP). The goal of our study was to investigate the potential for monitoring KRAS mutations during treatment with anti-EGFR antibodies. The detection limit of MBP-QP using a control plasmid containing KRAS mutations was 1-9 copies, and 0.05-0.3% mutant plasmid was detectable in a mixture of wild type and mutants. One-hundred twenty colorectal cancer patients were genotyped for KRAS mutations with MBP-QP as well as polymerase chain reaction reverse sequence-specific oligonucleotide (PCR-rSSO), which has already been applied to cancer tissue samples in the clinical setting. Concordance rates between plasma DNA and cancer tissues were 68% with MBP-QP and 66% with PCR-rSSO, indicating that these systems are equivalent in terms of detecting KRAS mutations with plasma DNA. KRAS mutations in plasma DNA were frequently observed in systemic metastatic cancer patients, and in three patients KRAS mutations appeared after chemotherapy containing anti-EGFR antibody. A prospective study is needed for clarifying whether KRAS mutations detected in plasma DNA are predictive markers of treatment efficacy with anti-EGFR antibody.
  • Yuji Kaneda, Hiroshi Noda, Yuhei Endo, Nao Kakizawa, Kosuke Ichida, Fumiaki Watanabe, Takaharu Kato, Yasuyuki Miyakura, Koichi Suzuki, Toshiki Rikiyama
    World journal of gastrointestinal oncology 9 9 372 - 378 2017年09月 [査読有り][通常論文]
     
    AIM: To assess the usefulness of en bloc right hemicolectomy with pancreaticoduodenectomy (RHCPD) for locally advanced right-sided colon cancer (LARCC). METHODS: We retrospectively reviewed the database of Saitama Medical Center, Jichi Medical University, between January 2009 and December 2016. During this time, 299 patients underwent radical right hemicolectomy for right-sided colon cancer. Among them, 5 underwent RHCPD for LARCC with tumor infiltration to adjacent organs. Preoperative computed tomography (CT) was routinely performed to evaluate local tumor infiltration into adjacent organs. During the operation, we evaluated the resectability and the amount of infiltration into the adjacent organs without dissecting the adherent organs from the cancer. When we confirmed that radical resection was feasible and could lead to R0 resection, we performed RHCPD. The clinical data were carefully reviewed, and the demographic variables, intraoperative data, and postoperative parameters were recorded. RESULTS: The median age of the 5 patients who underwent RHCPD for LARCC was 70 years. The tumors were located in the ascending colon (three patients) and transverse colon (two patients). Preoperative CT revealed infiltration of the tumor into the duodenum in all patients, the pancreas in four patients, the superior mesenteric vein (SMV) in two patients, and tumor thrombosis in the SMV in one patient. We performed RHCPD plus SMV resection in three patients. Major postoperative complications occurred in 3 patients (60%) as pancreatic fistula (grade B and grade C, according to International Study Group on Pancreatic Fistula Definition) and delayed gastric empty. None of the patients died during their hospital stay. A histological examination confirmed malignant infiltration into the duodenum and/or pancreas in 4 patients (80%), and no patients showed any malignant infiltration into the SMV. Two patients were histologically confirmed to have tumor thrombosis in the SMV. All of the tumors had clear resection margins (R0). The median follow-up time was 77 mo. During this period, two patients with tumor thrombosis died from liver metastasis. The overall survival rates were 80% at 1 year and 60% at 5 years. All patients with node-negative status (n = 2) survived for more than seven years. CONCLUSION: This study showed that the long-term survival is possible for patients with LARCC if RHCPD is performed successfully, particularly in those with node-negative status.
  • Koichi Suzuki, Yuta Muto, Kosuke Ichida, Taro Fukui, Yuji Takayama, Nao Kakizawa, Takaharu Kato, Fumi Hasegawa, Fumiaki Watanabe, Yuji Kaneda, Rina Kikukawa, Masaaki Saito, Shingo Tsujinaka, Kazushige Futsuhara, Osamu Takata, Hiroshi Noda, Yasuyuki Miyakura, Hirokazu Kiyozaki, Fumio Konishi, Toshiki Rikiyama
    Oncology letters 14 2 1491 - 1499 2017年08月 [査読有り][通常論文]
     
    Morphological response is considered an improved surrogate to the Response Evaluation Criteria in Solid Tumors (RECIST) model with regard to predicting the prognosis for patients with colorectal liver metastases. However, its use as a decision-making tool for surgical intervention has not been examined. The present study assessed the morphological response in 50 patients who underwent chemotherapy with or without bevacizumab for initially un-resectable colorectal liver metastases. Changes in tumor morphology between heterogeneous with uncertain borders and homogeneous with clear borders were defined as an optimal response (OR). Patients were also assessed as having an incomplete response (IR), and an absence of marked changes was assessed as no response (NR). No significant difference was observed in progression-free survival (PFS) between complete response/partial response (CR/PR) and stable disease/progressive disease (SD/PD), according to RECIST. By contrast, PFS for OR/IR patients was significantly improved compared with that for NR patients (13.2 vs. 8.7 months; P=0.0426). Exclusion of PD enhanced the difference in PFS between OR/IR and NR patients (15.1 vs. 9.3 months; P<0.0001), whereas no difference was observed between CR/PR and SD. The rate of OR and IR in patients treated with bevacizumab was 47.4% (9/19), but only 19.4% (6/31) for patients that were not administered bevacizumab. Comparison of the survival curves between OR/IR and NR patients revealed similar survival rates at 6 months after chemotherapy, but the groups exhibited different survival rates subsequent to this period of time. Patients showing OR/IR within 6 months appeared to be oncologically stable and could be considered as candidates for surgical intervention, including rescue liver resection. Comparing the pathological and morphological features of the tumor with representative optimal response, living tumor cells were revealed to be distributed within the area of vascular reconstruction induced by bevacizumab, resulting in a predictive value for prognosis in the patients treated with bevacizumab. The present findings provided the evidence for physicians to consider patients with previously un-resectable metastatic colorectal cancer as candidates for surgical treatment. Morphological response is a useful decision-making tool for evaluating these patients for rescue liver resection following chemotherapy.
  • Taro Fukui, Koichi Suzuki, Kosuke Ichida, Yuji Takayama, Nao Kakizawa, Yuta Muto, Fumi Hasegawa, Fumiaki Watanabe, Rina Kikugawa, Masaaki Saito, Shingo Tsujinaka, Yasuyuki Miyakura, Toshiki Rikiyama
    Oncology letters 13 6 4947 - 4952 2017年06月 [査読有り][通常論文]
     
    Sequential administration of the chemotherapy regimes capecitabine and oxaliplatin (XELOX) and capecitabine and irinotecan (XELIRI) in the first- to second-line treatment setting would allow patients to be managed more easily in an outpatient unit. However, a small number of studies have raised concerns of cumulative adverse events as a consequence of the continuous use of capecitabine. To investigate this, the present study conducted a retrospective review of 81 consecutive metastatic colorectal cancer (mCRC) patients treated with the oxaliplatin, fluorouracil and leucovorin-irinotecan, fluorouracil and leucovorin (FOLFOX-FOFIRI/F-F) regimen (n=40) or the XELOX-XELIRI (X-X) regimen (n=41) in first- to second-line chemotherapy in Saitama Medical Center between 2006 and 2012. The disease control rate (DCR), the progression free survival (PFS), the overall survival (OS) and the time to failure of strategy (TFS) from first to second-line chemotherapy, as well as adverse events, were assessed and compared between patients receiving X-X or F-F. A total of 10 and 20 patients were additionally treated with bevacizumab in the F-F and X-X regimens, respectively, during first or second-line chemotherapy. There was no significant difference in DCR and the median PFS between the two regimens for first or second-line chemotherapy. There was no significant difference in the median OS and TFS between the two regimens (OS=24.5 and TFS=14 months in the F-F vs. 23.2 and 12.0 months in the X-X). Regarding adverse events, 45.0% of patients (18/40) exhibited grade 3-4 neutropenia throughout treatment with F-F. Whilst, 15.0% of patients (6/41) exhibited grade 3 hypertension throughout treatment with X-X, which was effectively controlled by a single antihypertensive drug. The results show that sequential administration of X-X is as effective and feasible as F-F treatment, while additionally reducing the frequency of infusion visits and eliminating the need for a central venous access device or home infusion pump, thereby offering a more convenient treatment option to patients with mCRC.
  • Junji Mitsushita, Sachiho Netsu, Koichi Suzuki, Mitsuhiro Nokubi, Akira Tanaka
    International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists 36 3 253 - 260 2017年05月 [査読有り][通常論文]
     
    Approximately 1.6% of tumors metastatic to the ovary of nongynecologic origin are from a small bowel adenocarcinoma (SBA). However, the incidence of SBA is extremely rare (0.23 cases/100,000 people), which suggests a high frequency of ovarian metastasis, although the reason is unknown. To identify the characteristics of ovarian tumor metastasis from SBA, we reviewed 72 cases reported in the English literature, including the case presented in this report. The mean age of the patients was 46.7 yr. Solitary ovarian metastasis was observed in 67% of the cases, and ovarian metastasis was accompanied by peritoneal dissemination in 33% of the cases. Although duodenal adenocarcinoma has the highest incidence among the SBAs, jejunal adenocarcinoma, particularly that at the proximal end, is the type of SBA that most frequently metastasizes to the ovary. Among the cases of ovarian metastasis from SBA, 51% were bilateral, 33% were unilateral to the right ovary, and 16% were unilateral to the left ovary.
  • Nao Kakizawa, Koichi Suzuki, Taro Fukui, Yuji Takayama, Kosuke Ichida, Yuta Muto, Fumi Hasegawa, Fumiaki Watanabe, Rina Kikugawa, Shingo Tsujinaka, Kazushige Futsuhara, Yasuyuki Miyakura, Hiroshi Noda, Toshiki Rikiyama
    Oncology reports 37 4 2506 - 2512 2017年04月 [査読有り][通常論文]
     
    Regorafenib has shown survival benefits in metastatic colorectal cancer patients who were exacerbated after all standard therapies. Some patients, however, exhibit severe adverse events (AEs) resulting in treatment discontinuation. Therefore, the selection of patients likely to benefit from regorafenib is crucial. Twenty patients were treated with regorafenib for metastatic colorectal cancer; 122 plasma samples were taken from 16 of these patients for monitoring of circulating tumor DNA (ctDNA) in the blood. The treatment response, AEs, overall survival (OS), progression-free survival (PFS) and tumor morphologic changes on CT images were evaluated. KRAS mutant ctDNA was determined using digital PCR. Median PFS and OS were 2.5 and 5.9 months, respectively. Treatment was discontinued because of disease progression (PD) in 10 patients, and AEs in another 10 patients. AEs included hyperbilirubinemia, severe fatigue and skin rash. Hyperbilirubinemia was seen in two patients with multiple bilateral liver metastases, and severe fatigue in another 2 patients with poor performance status (PS). These severe AEs resulted in treatment discontinuation. Ten patients had a median PFS of 2.1 months with AE related discontinuation; PD occurred at 3.5 months (p=0.00334). Four patients exhibited a morphologic response, achieving better PFS times of 3.5, 5.3, 5.6 and 14.2 months. Emergence of the KRAS mutation in ctDNA was observed during anti-EGFR antibody treatment in 3 patients among 11 with KRAS wild-type tumors; it was detectable in the blood prior to radiographic detection of PD. Moreover, the KRAS mutation declined in two patients during regorafenib monotherapy. These patients were re-challenged with anti-EGFR antibody. Patients with extensive multiple liver metastases or poor PS are unlikely to benefit from regorafenib. Patients with a morphologic response will probably benefit from regorafenib with adequate management of other AEs. KRAS monitoring in ctDNA could be useful regarding treatment response and in determining treatment strategy.
  • Clinical characteristics of synchronous colorectal cancers in Japan.
    Takaharu Kato, Sergio Alonso, Yuta Muto, Hiroshi Noda, Yasuyuki Miyakura, Koichi Suzuki, Shingo Tsujinaka, Masaaki Saito, Manuel Perucho, Toshiki Rikiyama
    World journal of surgical oncology 14 1 272 - 272 2016年10月 [査読有り][通常論文]
     
    BACKGROUND: Incidence and clinical characteristics of synchronous colorectal cancer (sCRC) patients significantly vary among studies, likely due to differences in surveillance methodology. If remain undetected, sCRC can progress to more advanced stages seriously aggravating patient prognosis. We studied the incidence and clinicopathological characteristics of Japanese patients with sCRCs who underwent surgery for primary CRC and received exhaustive perioperative surveillance. METHODS: We recruited 1005 patients with surgically resected CRCs between January 2007 and December 2011. The associations of clinical and pathological factors with sCRC development were assessed by univariate and multivariate logistic regression. RESULTS: Eighty-four patients (8.4 %) developed sCRCs, 16 of them (19.0 %) harboring three or more cancers. Companion sCRCs were smaller and earlier stage than the index lesion (P < 0.0001). In multivariate analysis, advanced age (odds ratio (OR) 1.03 per year; P = 0.009) and left colon tumor location (OR 1.78; P = 0.013) are associated with higher risk of sCRCs, particularly in females. Overall survival did not differ between solitary CRC and sCRC (P = 0.62). CONCLUSIONS: Our results highlight the importance of perioperative colonoscopy examination to ensure the absence of sCRCs that, being small and early staged, are more difficult to detect. The incidence of sCRC, and notably of triple or more sCRCs, was higher than previously recognized. Because they are also significantly higher than expected by merely stochastic accumulation of individual cancerous lesions, we suggest that the occurrence of many sCRC reflects a hitherto uncharacterized predisposition condition.
  • Yuta Muto, Koichi Suzuki, Takaharu Kato, Shingo Tsujinaka, Kosuke Ichida, Yuji Takayama, Taro Fukui, Nao Kakizawa, Fumiaki Watanabe, Masaaki Saito, Kazushige Futsuhara, Hiroshi Noda, Yasuyuki Miyakura, Fumio Konishi, Toshiki Rikiyama
    International journal of oncology 49 3 1057 - 67 2016年09月 [査読有り][通常論文]
     
    Although epithelial-mesenchymal transition (EMT) has been implicated as the pivotal event in metastasis, there is insufficient evidence related to EMT in clinical settings. Intratumor heterogeneity may lead to underestimation of gene expression representing EMT. In the present study, we investigated the expression of EMT-associated genes and microRNAs in primary colorectal cancer while considering intratumor heterogeneity. One-hundred and thirty-three multiple spatially separated samples were obtained from 8 patients with metastatic colorectal cancers and 8 with non-metastatic colorectal cancers, from the tumor center (TC), invasive front (IF) and metastasis. Differences in gene and microRNA expression were investigated by microarray and quantitative reverse-transcription PCR. Gene expression microarray analysis detected 7920 sites showing differing levels of gene expression among the TC, IF and metastasis. Expression of the EMT-associated gene zinc-finger E-box-binding homeobox 1 (ZEB1) significantly increased in the IF (p<0.01). To exclude individual differences, the expression ratio between TC and IF in each tumor was applied to analysis. This approach enabled recognition of the activation of the VEGF and Wnt signaling pathways, which were involved in metastasis via promotion of EMT. While no activation of these pathways was seen at the TC, regardless of whether tumors were metastatic or non-metastatic, they were preferentially activated at the IF in metastatic tumors, where high ZEB1 expression was seen in connection with decreased miR-200c expression. Multiple sampling in a tumor revealed that heterogeneous ZEB1 expression induced by EMT-associated signaling pathways played a pivotal role in metastasis via regulation of miR-200c.
  • Takaharu Kato, Koichi Suzuki, Yuta Muto, Junichi Sasaki, Shingo Tsujinaka, Yutaka J Kawamura, Hiroshi Noda, Hisanaga Horie, Fumio Konishi, Toshiki Rikiyama
    World journal of surgical oncology 13 23 - 23 2015年02月 [査読有り][通常論文]
     
    BACKGROUND: Improvement in the prognosis of colorectal cancer (CRC) patients has led to increasing occurrences of multiple primary malignancies (MPMs) alongside CRC but little is known about their characteristics. This study was undertaken to clarify the clinical and pathological features of MPMs, especially those at extra colonic sites, in patients with CRC. METHODS: We reviewed 1,111 patients who underwent operations for primary sporadic CRC in Saitama Medical Center, Jichi Medical University between April 2007 and March 2012. Two patients with familial adenomatous polyposis, one with hereditary non-polyposis colorectal cancer, two with colitic cancer, and any patients with metastasis from CRC were excluded. We compared the clinicopathological features of CRC patients with and without MPMs. As a control, we used a database compiled of patients with gastric cancer (GC) detected by mass screening performed in the Saitama Prefecture in Japan 2010 and compared these with CRC patients with synchronous GC. RESULTS: Multiple primary malignancies at extracolonic sites were identified in 117 of 1,111 CRC patients (10.5%). The median age was 68 (range, 29 to 96) versus 71 (50 to 92) (P < 0.001). The incidence of GC (44.4% (52 of 117)) was the highest of all MPMs. All CRC patients with GC were older than 57 years. Synchronous GC was detected in 26 patients. By contrast, out of 200,007 screened people, 225 people were diagnosed as having GC in the Saitama Prefecture. The age-standardized incidence of synchronous GC in CRC patients was significantly higher (0.53%) than in the control group (0.03%) (odds ratio, 18.8; 95% confidence interval, 18.6 to 19.0; P < 0.001). CONCLUSION: Patients with CRC who were older than 50 years preferentially developed GC synchronously and metachronously. Thus, this patient group should undergo careful perioperative screening for GC.
  • K Tago, M Funakoshi-Tago, H Itoh, Y Furukawa, J Kikuchi, T Kato, K Suzuki, K Yanagisawa
    Oncogene 34 3 314 - 22 2015年01月 [査読有り][通常論文]
     
    Tumor suppressor protein p19(ARF) (Arf; p14(ARF) in humans) functions in both p53-dependent and -independent modes to counteract hyper-proliferative signals caused by proto-oncogene activation, but its p53-independent activities remain poorly understood. Using the tandem affinity purification-tag technique, we purified Arf-containing protein complexes and identified p68 DEAD-box protein (DDX5) as a novel interacting protein of Arf. In this study, we found that DDX5 interacts with c-Myc, and harbors essential roles for c-Myc-mediated transcription and its transforming activity. Furthermore, when c-Myc was forcibly expressed, the expression level of DDX5 protein was drastically increased through the acceleration of protein synthesis of DDX5, suggesting the presence of an oncogenic positive feedback loop including c-Myc and DDX5. Strikingly, Arf blocked the physical interaction between DDX5 and c-Myc, and drove away DDX5 from the promoter of c-Myc target genes. These observations most likely indicate the mechanism by which Arf causes p53-independent tumor-suppressive activity.
  • Sergio Alonso, Beatriz González, Tatiana Ruiz-Larroya, Mercedes Durán Domínguez, Takaharu Kato, Akihiro Matsunaga, Koichi Suzuki, Alex Y Strongin, Pepita Gimènez-Bonafé, Manuel Perucho
    Clinical epigenetics 7 124 - 124 2015年 [査読有り][通常論文]
     
    BACKGROUND: ADAMTS19 encodes a member of the ADAMTS (a disintegrin and metalloproteinase domain with thrombospondin motifs) protein family with emerging roles in carcinogenesis and metastasis. ADAMTS shares several distinct protein modules including a propeptide region, a metalloproteinase domain, a disintegrin-like domain, and a thrombospondin type 1 (TS) motif. In a previous work, we found ADAMTS19 frequently hypermethylated in colorectal cancer (CRC). We explored the association of methylation with tumor genotype and phenotype. RESULTS: The methylation status of the CpG island in the promoter of ADAMTS19 was determined in 252 colorectal, 65 pancreatic, 33 breast and 169 ovarian primary tumors, 70 CRC metastases, and 10 CRC cell lines. Tumor-specific methylation of ADAMTS19 was significantly more frequent in gastrointestinal than in gynecological cancers (odds ratio (OR) = 2.9, confidence interval (CI) = (1.9-4.7), p = 5.2 × 10(-7)) and was independent of the methylation of adjacent loci in CRC. Hypermethylation associated with CRC with mutated BRAF oncogene (OR = 10.1, CI = (3.1-42.9), p = 6.3 × 10(-6)) and with the mucinous phenotype in CRC (OR = 2.1, CI = (1.1-4.1), p = 0.023) and ovarian cancer (OR = 60, CI = (16-346), p = 4 × 10(-16)). Methylation was significantly more frequent in CRC metastases homing to the ovary and omentum than in those homing to the liver and lung (OR = 6.1, CI = (1.8-22.2), p = 0.001). Differentiating local from distant metastatic spread, methylation negatively associated with tumor progression (p = 0.031) but positively with depth of invasion (p = 0.030). Hypermethylation associated with transcriptional repression in CRC cell lines, and treatment with 5'-AZA-2'-deoxycytidine led to reactivation of mRNA expression. shRNA-mediated silencing of ADAMTS19 had no effect on the in vitro proliferation rate of CRC cells but significantly diminished their collective migration speed (56 %, p = 3.3 × 10(-4)) and potential to migrate in collagen I (64 %, p = 4.3 × 10(-10)). CONCLUSIONS: Our results highlight the frequent involvement of ADAMTS19 epigenetic silencing in CRC and mucinous ovarian cancer. The mechanistic preferences for the target organ of metastatic spread may lead to the development of diagnostic CRC biomarkers. The association with the mucinous phenotype also may have diagnostic applications for ovarian cancer.
  • Masaaki Saito, Hirokazu Kiyozaki, Osamu Takata, Koichi Suzuki, Toshiki Rikiyama
    World journal of surgical oncology 12 406 - 406 2014年12月 [査読有り][通常論文]
     
    BACKGROUND: The standard treatment for stage IV gastric cancer is chemotherapy, but outcomes remain poor. The effectiveness of induction chemotherapy followed by surgery in selected patients who had a good response to chemotherapy is unclear. METHODS: A total of 59 patients with stage IV gastric cancer received induction chemotherapy with S-1 and cisplatin. In each cycle, oral S-1 (80 mg/m2) was administered for 3 weeks, followed by a 2-week drug holiday. Intravenous cisplatin (60 mg/m2) was administered on day 8 after adequate premedication and hydration. If unresectable features resolved after chemotherapy, patients underwent curative (R0) resection. The safety and outcomes of this treatment combination were evaluated, and predictive factors for survival were determined. RESULTS: Thirteen of 59 patients (22%) were eligible for R0 resection after induction chemotherapy. Kaplan-Meier analysis showed an overall median survival time of 13 months and a 3-year survival rate of 18.2%. Among patients who underwent R0 resection, the median survival time was 53 months and the 3-year survival rate was 53.8%. Multivariate analyses showed that negative para-aortic lymph nodes and undergoing R0 resection were independent predictors of survival. CONCLUSIONS: Treatment of stage IV gastric cancer with S-1 and cisplatin induction chemotherapy followed by R0 resection is safe and may improve survival compared with chemotherapy alone. Further study of this dual-modality therapy is warranted.
  • XELIRI regimen plus continuous treatment with bevacizumab is well-tolerated and effective in metastatic colorectal cancer patients in a second-line setting involving the sequential administration of XELOX and XELIRI.
    Koichi Suzuki, Kato Takaharu, Yuta Muto, Kosuke Ichida, Taro Fukui, Yuji Takayama, Shingo Tsujinaka, Junichi Sasaki, Hisanaga Horie, Yutaka J Kawamura, Fumio Konishi, Toshiki Rikiyama
    Molecular and clinical oncology 2 5 827 - 832 2014年09月 [査読有り][通常論文]
     
    The aim of the present study was to present a retrospective review of 42 metastatic colorectal cancer (mCRC) patients treated using the XELIRI regimen as second-line chemotherapy during the period between 2010 and 2012. Patients were treated with capecitabine, 1,600 (≥65 years) or 2,000 mg/m2 (<65 years), on days 1-15, 200 mg/m2 irinotecan (CPT-11) on day 1, with or without 7.5 mg/kg bevacizumab on day 1 and every 21 days. A total of 21 patients underwent XELIRI and 21 underwent XELIRI plus bevacizumab treatment. Fifteen patients received continuous administration of bevacizumab in the first- and second-line settings [bevacizumab beyond progression (BBP)+], whereas 27 patients did not receive the treatment (BBP-). Forty patients (95.2%), including all the patients in the BBP+ group, received sequentially administered XELOX and XELIRI regimens from the first- to the second-line setting. The disease control rate (DCR), progression-free survival (PFS), overall survival (OS) and adverse events were compared between the BBP- and BBP+ groups. The median relative dose intensity was similar (93.9% for capecitabine and 96.3% for CPT-11 in the BBP- group vs. 94.8% for capecitabine and 91.5% for CPT-11 in the BBP+ group). The DCR was 25.9% in the BBP- and 66.6% in the BBP+ groups (P=0.020). The median PFS was 3.5 months in the BBP- and 7.2 months in the BBP+ groups (P=0.028). The BBP+ group exhibited a higher median OS time compared to the BBP- group (12.5 months in the BBP- group vs. not reached in the BBP+ group; P=0.0267). The most common grade 3/4 adverse event (n≥20) was hypertension observed in the BBP+ group [three patients (20%)]: these three patients were well-controlled with a single antihypertensive drug. Treatment with sequentially administered XELOX and XELIRI regimens did not aggravate adverse events in the 40 patients. The results showed that the XELIRI regimen, involving continuous treatment with bevacizumab, was well-tolerated and effective as a second-line chemotherapy and sequentially administering XELOX and XELIRI was feasible and manageable for patients with mCRC.
  • Yuta Muto, Takafumi Maeda, Koichi Suzuki, Takaharu Kato, Fumiaki Watanabe, Hidenori Kamiyama, Masaaki Saito, Kei Koizumi, Yuichiro Miyaki, Fumio Konishi, Sergio Alonso, Manuel Perucho, Toshiki Rikiyama
    BMC cancer 14 466 - 466 2014年06月 [査読有り][通常論文]
     
    BACKGROUND: Recent work led to recognize sessile serrated adenomas (SSA) as precursor to many of the sporadic colorectal cancers with microsatellite instability (MSI). However, comprehensive analyses of DNA methylation in SSA and MSI cancer have not been conducted. METHODS: With an array-based methylation sensitive amplified fragment length polymorphism (MS-AFLP) method we analyzed 8 tubular (TA) and 19 serrated (SSA) adenomas, and 14 carcinomas with (MSI) and 12 without (MSS) microsatellite instability. MS-AFLP array can survey relative differences in methylation between normal and tumor tissues of 9,654 DNA fragments containing all NotI sequences in the human genome. RESULTS: Unsupervised clustering analysis of the genome-wide hypermethylation alterations revealed no major differences between or within these groups of benign and malignant tumors regardless of their location in intergenic, intragenic, promoter, or 3' end regions. Hypomethylation was less frequent in SSAs compared with MSI or MSS carcinomas. Analysis of variance of DNA methylation between these four subgroups identified 56 probes differentially altered. The hierarchical tree of this subset of probes revealed two distinct clusters: Group 1, mostly composed by TAs and MSS cancers with KRAS mutations; and Group 2 with BRAF mutations, which consisted of cancers with MSI and MLH1 methylation (Group 2A), and SSAs without MLH1 methylation (Group 2B). AXIN2, which cooperates with APC and β-catenin in Wnt signaling, had more methylation alterations in Group 2, and its expression levels negatively correlated with methylation determined by bisulfite sequencing. Within group 2B, low and high AXIN2 expression levels correlated significantly with differences in size (P = 0.01) location (P = 0.05) and crypt architecture (P = 0.01). CONCLUSIONS: Somatic methylation alterations of AXIN2, associated with changes in its expression, stratify SSAs according to some clinico-pathological differences. We conclude that hypermethylation of MLH1, when occurs in an adenoma cell with BRAF oncogenic mutational activation, drives the pathway for MSI cancer by providing the cells with a mutator phenotype. AXIN2 inactivation may contribute to this tumorigenic pathway either by mutator phenotype driven frameshift mutations or by epigenetic deregulation contemporary with the unfolding of the mutator phenotype.
  • H Kamiyama, K Suzuki, T Maeda, K Koizumi, Y Miyaki, S Okada, Y J Kawamura, J K Samuelsson, S Alonso, F Konishi, M Perucho
    Oncogene 31 48 5029 - 37 2012年11月 [査読有り][通常論文]
     
    Some colon cancer (CC) patients present synchronous cancers at diagnosis and others develop metachronous neoplasms, but the risk factors are unclear for non-hereditary CC. We showed previously that global DNA demethylation increased with aging and correlated with genomic damage in CC, and we show now that preferentially associates to CCs with wild-type p53. This study aimed to elucidate the extent of DNA hypomethylation in patients with single and multiple CC, its relationship with aging, and its potential as predictive tool. We compared by real-time methylation-specific PCR the relative demethylation level (RDL) of long interspersed nucleotide element-1 (LINE-1) sequences in matched cancer tissues and non-cancerous colonic mucosa (NCM) from patients with single and multiple right-sided CCs. Although no RDL difference was found in NCM from single CC patients and healthy volunteers (P=0.5), there was more demethylation (higher RDL) in NCM from synchronous cancer patients (P=1.1 × 10(-5)) multiple CCs also were more demethylated than single CCs (P=0.0014). High NCM demethylation was predictive for metachronous neoplasms (P=0.003). In multivariate logistic regression analyses RDL was the only independent predictor for metachronous (P=0.02) and multiple (P=4.9 × 10(-5)) tumors. The higher LINE-1 demethylation in NCM from patients with multiple (synchronous and metachronous) tumors (P=9.6 × 10(-7)) was also very significant in patients with tumors without (P=3.8 × 10(-6)), but not with (P=0.16) microsatellite instability. NCM demethylation increased with aging in patients with single tumors, but decreased in those with multiple tumors. Moreover, the demethylation difference between patients with single vs multiple tumors appeared higher in younger (P=3.6 × 10(-4)) than in older (P=0.0016) patients. These results predict that LINE-1 hypomethylation in NCM can be used as an epigenetic predictive biomarker for multiple CC risk. The stronger association of demethylation in NCM with multiple CC risk from younger patients also suggests an inherited predisposition for the apparent field cancerization effect of somatic demethylation.
  • Masaaki Saito, Koichi Suzuki, Takafumi Maeda, Takaharu Kato, Hidenori Kamiyama, Kei Koizumi, Yuichiro Miyaki, Shinichiro Okada, Hirokazu Kiyozaki, Fumio Konishi
    Oncology reports 27 6 1717 - 25 2012年06月 [査読有り][通常論文]
     
    Helicobacter pylori (HP) infection is widely recognized as a risk factor for gastric cancer, but only a minority of infected individuals develop gastric cancer. The aim of this study was to determine whether DNA demethylation in non-cancerous gastric mucosa (NGM) significantly enhances susceptibility to gastric cancer. A total of 165 healthy volunteers, including 83 HP-positive and 82-negative individuals, as well as 83 patients with single and 18 with synchronous double gastric cancer (GC) were enrolled in this study. The relative demethylation levels (RDLs) of repetitive sequences, including Alu, LINE-1 and Sat α, were quantified by real-time methylation-specific polymerase chain reaction. The Alu RDL did not exhibit any differences within each respective group, whereas LINE-1 RDL was significantly elevated in cancer tissues compared with the NGM in the other groups (P<0.001). Our results indicated that a gradual increase in Sat α RDL correlated with HP infection and cancer development. Sat α RDL was significantly elevated in the NGM in HP-positive compared with HP-negative (P<0.001), and significantly elevated in cancer tissues (P<0.001). Although the Sat α RDL of the NGM in the total population increased in an age-dependent manner, it was significantly increased in a fraction of younger GC patients (<45 years) compared with all of the others (45 years or older, P=0.0391). In addition, double GC exhibited a significantly higher Sat α RDL in the NGM compared with single GC (P=0.0014). In these two fractions, Sat α RDL in the NGM exhibited an inverse correlation with age. In conclusion, the present study demonstrated that the accumulation of DNA demethylation in Sat α RDL in the NGM with HP infection potentially renders susceptibility to gastric cancer in a fraction of GC patients younger than 45 years or in patients with multiple cancers.
  • Kei Koizumi, Sergio Alonso, Yuichiro Miyaki, Shinichiro Okada, Hiroyuki Ogura, Norihiko Shiiya, Fumio Konishi, Toshiki Taya, Manuel Perucho, Koichi Suzuki
    International journal of oncology 40 4 983 - 94 2012年04月 [査読有り][通常論文]
     
    Patients with long-standing ulcerative colitis (UC) have higher risk of developing colorectal cancer. Albeit the causes remain to be understood, epigenetic alterations have been suggested to play a role in the long-term cancer risk of these patients. In this work, we developed a novel microarray platform based on methylation-sensitive amplified fragment length polymorphism (MS-AFLP) DNA fingerprinting. The over 10,000 NotI sites of the human genome were used to generate synthetic primers covering these loci that are equally distributed into CpG rich regions (promoters and CpG islands) and outside the CpG islands, providing a panoramic view of the methylation alterations in the genome. The arrays were first tested using the colon cancer cell line CW-2 showing the reproducibility and sensitivity of the approach. We next investigated DNA methylation alterations in the colonic mucosa of 14 UC patients. We identified epigenetic alterations affecting genes putatively involved in UC disease, and in susceptibility to develop colorectal cancer. There was a strong concordance of methylation alterations (both hypermethylation and hypomethylation) shared by the cancer cells of the CW-2 cell line and the non-cancer UC samples. To the best of our knowledge, this work defines the first high-throughput aberrant DNA methylation profiles of the colonic mucosa of UC patients. These epigenetic profiles provide novel and relevant knowledge on the molecular alterations associated to the UC pathology. Some of the detected alterations could be exploited as cancer risk predictors underlying a field defect for cancerization in UC-associated carcinogenesis.
  • Takaharu Kato, Koichi Suzuki, Shinichiro Okada, Hidenori Kamiyama, Takafumi Maeda, Masaaki Saito, Kei Koizumi, Yuichiro Miyaki, Fumio Konishi
    International journal of oncology 40 4 942 - 50 2012年04月 [査読有り][通常論文]
     
    We previously reported that the Pleckstrin and Sec7 domain-containing (PSD) gene is preferentially methylated in patients with ulcerative colitis (UC) who developed colorectal cancer (CRC), and is implicated in UC-associated carcinogenesis through its inhibition of apoptosis. This study aimed to determine the potential effect of PSD methylation on its downstream molecule, Ras-related C3 botulinum toxin substrate 1 (Rac1), which governs neutrophil chemotaxis and apoptosis signaling. PSD was knocked down in a normal human fibroblast cell line (HNDF) and a neutrophil-like cell line (HL-60). Both NHDF and HL-60 cells exhibited numerous filamentous-actin (F-actin) rich membrane extensions, resulting in the activation of Rac1; this activation was hampered by PSD silencing. Lipopolysaccharide, a reactive oxygen species (ROS) inducer, stimulated NHDF cells to release ROS and activated caspase‑3/7 in the presence of neutrophils, which was inhibited by PSD knockdown. Migration assays demonstrated that chemotaxis of HL-60 cells was affected by PSD silencing in NHDF cells. Tissue sections from 6 UC patients with CRC and 15 UC patients without CRC were examined. To verify Rac1-mediated chemotaxis in tissue sections, we evaluated the grade of neutrophil infiltration by histological assessment and assessed F-actin and PSD expression by immunohistochemistry. Neutrophil infiltration, F-actin and PSD expression were significantly decreased in specimens from UC patients with PSD methylation compared with those without. Decreased levels of F-actin expression were observed in colorectal mucosa, as well as in infiltrating cells with PSD methylation. PSD expression was preferentially inhibited in colorectal mucosa by PSD methylation, whereas PSD expression was rarely observed in infiltrating cells, regardless of PSD methylation status. These data indicate that aberrant methylation of PSD occurs in UC-associated colorectal mucosa, enabling circumvention of Rac1-mediated immune responses governing neutrophil chemotaxis and apoptosis, and thus plays a pivotal role in the mechanisms underlying UC-associated carcinogenesis.
  • Shinichiro Okada, Koichi Suzuki, Kato Takaharu, Hiroshi Noda, Hidenori Kamiyama, Takafumi Maeda, Masaaki Saito, Kei Koizumi, Yuichiro Miyaki, Fumio Konishi
    International journal of oncology 40 3 686 - 94 2012年03月 [査読有り][通常論文]
     
    The Pleckstrin and Sec7 domain-containing (PSD) gene, which regulates skeletal rearrangements, has been found to be more frequently methylated both in ulcerative colitis (UC)-associated colorectal cancer tissues (5 of 7; 71.4%) and matched normal epithelia (4 of 7; 57.1%) compared to non-neoplastic UC epithelia (6 of 22; 27.3%) and sporadic colorectal cancer tissues (6 of 32; 18.8%). The levels of PSD mRNA were positively correlated with the methylation status of PSD, as shown by both MSP and bisulfite sequencing. To determine the potential role of PSD silencing in the mechanisms underlying UC-associated carcinogenesis, the levels of senescence, proliferation and apoptosis were evaluated in a normal human fibroblast cell line (NHDF) in which 93% of PSD expression was knocked down by a small-interfering RNA (si-RNA). Although there were no significant differences in the levels of senescence and proliferation caused by PSD knockdown, the level of apoptosis was significantly decreased by PSD knockdown (5.3% in siControl-treated cells vs. 0.67% in siPSD-treated cells, p=0.0001). In addition, reactive oxygen species inducers accelerated apoptosis in NHDF and a neutrophil-like cell line, which was significantly reduced by PSD knockdown. To verify the effect of PSD methylation in tissue sections including 21 samples from UC patients with or without tumors, we elucidated PSD promoting accumulation of filamentous-actin (F-actin) and apoptosis by immunohistochemistry and TUNEL assay, respectively. Both levels of accumulation of F-actin and apoptosis were significantly decreased in specimens from UC patients with PSD methylation compared to those without PSD methylation (F-actin: 0.69±0.86 with vs. 1.57±0.51 without, p=0.0031, apoptotic index: 0.31±0.63 with vs. 1.0±0.88 without, p=0.0277). In conclusion, our results indicate that PSD methylation plays a significant role in the mechanisms underlying UC-associated carcinogenesis through its inhibitory effect on apoptosis in the interaction between colorectal mucosa and neutrophils.
  • Yuichiro Miyaki, Koichi Suzuki, Kei Koizumi, Takaharu Kato, Masaaki Saito, Hidenori Kamiyama, Takafumi Maeda, Kiyoshi Shibata, Norihiko Shiya, Fumio Konishi
    International journal of oncology 40 1 217 - 26 2012年01月 [査読有り][通常論文]
     
    Drug resistance remains a major obstacle to successful cancer treatment. Genome-wide comprehensive analysis identified a novel gene, glucocorticoid-induced protein-coding gene (DEXI), which was frequently methylated in colorectal (CRC; 36 of 73 patients; 49%) and gastric (28 of 89 patients; 31%) cancer patients. Here, we show that DEXI methylation is implicated in mechanisms facilitating resistance to camptothecin (CPT) via inhibition of apoptosis. Silencing of DEXI by siRNA significantly reduced CPT-induced apoptosis in a fibroblast cell line (1/6-fold; p<0.01) originally expressing endogenous DEXI. Restored expression of DEXI by 5-aza-2'-deoxycytidine (DAC) significantly enhanced susceptibility to CPT (3-fold; p<0.01) in a colon cancer cell line originally suppressing endogenous DEXI due to almost complete methylation. Exogenous induction of DEXI confirmed that DEXI per se contributed to enhanced susceptibility to CPT. 5-Fluorouracil (5-FU) did not exhibit these synergistic effects by DEXI restoration. Further, to estimate the clinical usefulness of DEXI methylation status as biomarker for drug resistance to irinotecan (CPT-11), 16 CRC patients who underwent FOLFIRI (5-FU + CPT-11) therapy because they were refractory to FOLFOX (5-FU + oxaliplatin) were analyzed. Significantly poor response and outcome were observed in 8 CRC patients harboring DEXI methylation. In 8 CRC patients harboring DEXI methylation disease control rate, progression-free survival and overall survival were 25.0%, 2 and 11.8 months, respectively, whereas in 8 CRC patients without DEXI methylation they were 62.5%, 5.3 and 15 months, respectively (p<0.01). These significant differences were not observed in patients undergoing treatment with FOLFOX. In conclusion, silencing of DEXI leads to resistance, but restored expression enhances susceptibility to CPT in vitro and DEXI methylation results in poor response and outcome to CPT-11-based chemotherapy, suggesting that DEXI is a potent therapeutic target and an epigenetic biomarker for the selection of patients more likely to benefit from CPT-11-based chemotherapy.
  • Sessile serrated adenoma shares similar genetic and epigenetic features with microsatellite unstable colon cancer in a location-dependent manner.
    Takafumi Maeda, Koichi Suzuki, Kazutomo Togashi, Mitsuhiro Nokubi, Masaaki Saito, Shingo Tsujinaka, Hidenori Kamiyama, Fumio Konishi
    Experimental and therapeutic medicine 2 4 695 - 700 2011年07月 [査読有り][通常論文]
     
    Genetic and epigenetic features of sessile serrated adenoma (SSA), a precursor lesion to colon cancer with microsatellite instability (MSI), were investigated. The aim of this study was to clarify whether there are location-dependent genetic and epigenetic features in SSA. Twenty-two patients with proximal SSAs and 8 with distal SSAs were recruited. Twenty-two patients with tubular adenoma (TA) and 66 with proximal colon cancer were studied for comparison. Genetic and epigenetic features were evaluated as BRAF and KRAS mutations, MSI, hMLH1 methylation and CpG island methylator phenotype (CIMP). BRAF mutation (p=0.007) and CIMP (p=0.012) were more frequently found in proximal than in distal SSAs. Furthermore, the KRAS mutation was found only in distal SSAs. In TAs, no location-related molecular features were observed. All SSAs, TAs and 42 colon cancer lesions were microsatellite stable (MSS). Twenty-four colon cancer lesions exhibited MSI and had more frequent BRAF mutations (p<0.001), hMLH1 methylation (p<0.001) and CIMP (p<0.001). BRAF mutation occurred in only 9.5% of MSS cancers (p=0.01). In MSI cancers with the BRAF mutation, a higher correlation with CIMP (p=0.032) was observed. We demonstrated the distinct genetic and epigenetic features between proximal and distal SSAs. Similar genetic and epigenetic features were shared between proximal SSAs and proximal MSI cancers harboring the BRAF mutation. By contrast, our results allow the possibility of carcinogenesis in SSAs leading to MSS cancer with the BRAF mutation.
  • Kok-Yang Tan, Fumio Konishi, Koichi Suzuki
    Surgery today 40 4 385 - 7 2010年04月 [査読有り][通常論文]
     
    This article critically discusses the current evidence for adjuvant chemotherapy in elderly patients (> or =70 years of age) with stage III colon cancer. The authors emphasize that current evidence is inconclusive, and surgeons should be aware of this fact when making informed decisions and recommendations.
  • Hidenori Kamiyama, Hiroshi Noda, Osamu Takata, Koichi Suzuki, Yutaka Kawamura, Fumio Konishi
    Journal of surgical oncology 100 1 69 - 74 2009年07月 [査読有り][通常論文]
     
    BACKGROUND AND OBJECTIVES: The predictive value of free cancer cells in the peritoneal fluid of patients with colorectal cancer (CRC) remain to be elucidated. The aim of this study was to determine the prognostic relevance of the methylation of tumor-related genes detected in the peritoneal lavage fluid (PLF) of patients undergoing a resection for CRC. METHODS: The promoter methylation pattern of four target genes, CDH1, CDKN2A (p16), MGMT, and APC, was examined in 51 primary CRC and corresponding matched PLF DNA. The relative methylation levels of these genes in primary CRC tissue and paired PLF were assessed by quantitative methylation-specific polymerase chain reaction (QMSP). RESULTS: An aberrant methylation of at least one gene was found in 45 of 51 (88%) primary tumors. In matched PLF specimens, the frequencies of aberrant promoter methylation detected for each marker were 16% for CDH1, 2% for p16, 4% for MGMT and 24% for APC. Patients with PLF demonstrating the methylation of more than one of these four target genes demonstrated significantly shorter relapse-free survival. CONCLUSIONS: These findings suggest that disseminated tumor cells in PLF detected by QMSP may correlate with the postoperative clinical course of patients undergoing curative surgery for CRC.
  • Shinji Kageyama, Kazuya Shinmura, Hiroko Yamamoto, Masanori Goto, Koichi Suzuki, Fumihiko Tanioka, Toshihiro Tsuneyoshi, Haruhiko Sugimura
    Japanese journal of clinical oncology 38 4 317 - 22 2008年04月 [査読有り][通常論文]
     
    The PCR-based DNA fingerprinting method called the methylation-sensitive amplified fragment length polymorphism (MS-AFLP) analysis is used for genome-wide scanning of methylation status. In this study, we developed a method of fluorescence-labeled MS-AFLP (FL-MS-AFLP) analysis by applying a fluorescence-labeled primer and fluorescence-detecting electrophoresis apparatus to the existing method of MS-AFLP analysis. The FL-MS-AFLP analysis enables quantitative evaluation of more than 350 random CpG loci per run. It was shown to allow evaluation of the differences in methylation level of blood DNA of gastric cancer patients and evaluation of hypermethylation and hypomethylation in DNA from gastric cancer tissue in comparison with adjacent non-cancerous tissue.
  • How do we manage the gastrectomy for gastric cancer after coronary artery bypass grafting using the right gastroepiploic artery? Report of two cases and a review of the literature.
    Yukiko Konishi, Koichi Suzuki, Hidetoshi Wada, Hiroshi Watanabe, Hiroyuki Ogura, Yuno Sugamori, Abul Hasan Muhammad Bashar, Katsushi Yamashita, Toshihiko Kobayashi, Teruhisa Kazui
    World journal of surgical oncology 5 54 - 54 2007年05月 [査読有り][通常論文]
     
    BACKGROUND: Recently, the right gastroepiploic artery (RGEA) has been used in coronary artery bypass grafting (CABG) as an alternative arterial graft. Unfortunately, an increased incidence of gastric cancers has been reported after CABG using the RGEA. Handling of the RGEA during gastrectomy in these patients may cause lethal complications, which sometimes reduces the feasibility of curative dissection of lymph nodes at the base of the graft. CASE PRESENTATIONS: We describe two cases of gastric cancer undergoing gastrectomy after CABG with the use of RGEA. To avoid the potentially fatal coronary event during gastrectomy, safe handling of the conduit including preparations for injuries and prevention of vessel spasm was performed in both cases, accompanied by an adequate monitoring of the systemic circulation. Intraoperative frozen section examination showed no lymph node metastasis around the graft in any of the cases; therefore, complete lymph node dissection at the base of the graft was not undertaken. No complications occurred during the operation. In addition to these two cases, twenty-four cases reported in the literatures were reviewed (a total of 26 cases). Ten early and 16 advanced gastric cancers were included. Among the 16 advanced gastric cancer cases, an alternative graft was employed in 8 due to the resection of an original graft to complete lymph node dissection. Mere handling of a graft often caused lethal complications suggesting that the operation should be completed by isolation of the graft. A pedicled graft harvesting via the ante-gastric route was popular. However, a skeletonized harvesting with resection of the pyloric branches of the RGEA would be better because this would interrupt the original lymph flow, which could eliminate the need for lymph node dissection and graft isolation. Among the 10 cases having early gastric cancers, 6 were found within 1.5 years after CABG. Early detection in these 6 cases was possible due to the use of gastric fiberscopic examination before and after CABG, which gave them opportunities to receive a less extensive operation such as endoscopic mucosal resection. CONCLUSION: Adequate intraoperative care as well as an optimal lymph node dissection considering the graft harvesting method at the first CABG leads to successful gastrectomy after CABG using the RGEA graft. Therefore, this operation should be carried out with careful management by both gastrointestinal and cardiovascular surgeons.
  • Global DNA demethylation in gastrointestinal cancer is age dependent and precedes genomic damage.
    Koichi Suzuki, Ikuko Suzuki, Andreas Leodolter, Sergio Alonso, Shina Horiuchi, Kentaro Yamashita, Manuel Perucho
    Cancer cell 9 3 199 - 207 2006年03月 [査読有り][通常論文]
     
    We studied the relationships between genetic and epigenetic alterations in gastrointestinal cancer by integrating DNA copy number changes determined by arbitrarily primed PCR (AP-PCR) with DNA methylation variations estimated by methylation-sensitive amplified fragment length polymorphism (MS-AFLP). We analyzed about 100 different chromosomal regions by AP-PCR and over 150 random CpG loci by MS-AFLP in human colon and gastric carcinomas. DNA hypomethylation and hypermethylation alterations distributed gradually and increased with cancer patient age, in contrast with the age-independent genomic alterations. Increased DNA hypomethylation and hypermethylation correlated with increased genomic damage, but only hypomethylation was highly significant in multivariate analyses. We conclude that age-dependent accumulation of DNA demethylation precedes diploidy loss in a significant subset of gastrointestinal cancers.
  • Analysis of apoptosis protein expression in early-stage colorectal cancer suggests opportunities for new prognostic biomarkers.
    Maryla Krajewska, Hoguen Kim, Chul Kim, Haeyoun Kang, Kate Welsh, Shu-Ichi Matsuzawa, Michelle Tsukamoto, Ronald G Thomas, Nuria Assa-Munt, Zhe Piao, Koichi Suzuki, Manuel Perucho, Stan Krajewski, John C Reed
    Clinical cancer research : an official journal of the American Association for Cancer Research 11 15 5451 - 61 2005年08月 [査読有り][通常論文]
     
    PURPOSE: Although most stage II colon cancers are potentially curable by surgery alone, approximately 20% of patients relapse, suggesting a need for establishing prognostic markers that can identify patients who may benefit from adjuvant chemotherapy. We tested the hypothesis that differences in expression of apoptosis-regulating proteins account for differences in clinical outcome among patients with early-stage colorectal cancer. EXPERIMENTAL DESIGN: Tissue microarray technology was employed to assay the expression of apoptosis-regulating proteins by immunohistochemistry in 106 archival stage II colorectal cancers, making correlations with disease-specific survival. The influence of microsatellite instability (MSI), tumor location (left versus right side), patient age, and gender was also examined. RESULTS: Elevated expression of several apoptosis regulators significantly correlated with either shorter (cIAP2; TUCAN) or longer (Apaf1; Bcl-2) overall survival in univariate and multivariate analyses. These biomarkers retained prognostic significance when adjusting for MSI, tumor location, patient age, and gender. Moreover, certain combinations of apoptosis biomarkers were highly predictive of death risk from cancer. For example, 97% of patients with favorable tumor phenotype of cIAP2(low) plus TUCAN(low) were alive at 5 years compared with 60% of other patients (P = 0.00003). In contrast, only 37% of patients with adverse biomarkers (Apaf1(low) plus TUCAN(high)) survived compared with 83% of others at 5 years after diagnosis (P< 0.0001). CONCLUSIONS: Immunohistochemical assays directed at detection of certain combinations of apoptosis proteins may provide prognostic information for patients with early-stage colorectal cancer, and therefore could help to identify patients who might benefit from adjuvant chemotherapy or who should be spared it.
  • Interferon-alpha and antisense K-ras RNA combination gene therapy against pancreatic cancer.
    Kazuteru Hatanaka, Koichi Suzuki, Yoshiaki Miura, Kimiko Yoshida, Shumpei Ohnami, Yukio Kitade, Teruhiko Yoshida, Kazunori Aoki
    The journal of gene medicine 6 10 1139 - 48 2004年10月 [査読有り][通常論文]
     
    Interferon alpha (IFN-alpha) is used worldwide for the treatment of a variety of cancers. For pancreatic cancer, recent clinical trials using IFN-alpha in combination with standard chemotherapeutic drugs showed some antitumor activity of the cytokine, but the effect was not significant enough to enlist pancreatic cancer as a clinically effective target of IFN-alpha. In general, an improved therapeutic effect and safety are expected for cytokine therapy when given in a gene therapy context, because the technology would allow increased local concentrations of this cytokine in the target sites. In this study, we first examined the antiproliferative effect of IFN-alpha gene transduction into pancreatic cancer cells. The expression of IFN-alpha effectively induced growth suppression and cell death in pancreatic cancer cells, an effect which appeared to be more prominent when compared with other types of cancers and normal cells. Another strategy we have been developing for pancreatic cancer targets its characteristic genetic aberration, K-ras point mutation, and we reported that the expression of antisense K-ras RNA significantly suppressed the growth of pancreatic cancer cells. When these two gene therapy strategies are combined, the expression of antisense K-ras RNA significantly enhanced IFN-alpha-induced cell death (1.3- to 3.5-fold), and suppressed subcutaneous growth of pancreatic cancer cells in mice. Because the 2',5'-oligoadenylate synthetase/RNase L pathway, which is regulated by IFN and induces apoptosis of cells, is activated by double-strand RNA, it is plausible that the double-strand RNA formed by antisense and endogenous K-ras RNA enhanced the antitumor activity of IFN-alpha. This study suggested that the combination of IFN-alpha and antisense K-ras RNA is a promising gene therapy strategy against pancreatic cancer.
  • The genomic damage estimated by arbitrarily primed PCR DNA fingerprinting is useful for the prognosis of gastric cancer.
    Koichi Suzuki, Sumiko Ohnami, Chikako Tanabe, Hiroki Sasaki, Jun Yasuda, Hitoshi Katai, Kimio Yoshimura, Masaaki Terada, Manuel Perucho, Teruhiko Yoshida
    Gastroenterology 125 5 1330 - 40 2003年11月 [査読有り][通常論文]
     
    BACKGROUND & AIMS: Genomic instability and the accompanying alteration of cancer genes play a major role in tumorigenesis. We evaluated the prognostic significance in gastric cancer of the degree of accumulation of relative genomic damage, assessed by arbitrarily primed polymerase chain reaction DNA fingerprinting. METHODS: Genomic damage was assessed by comparative analysis of paired normal and tumor tissue DNA fingerprints. The total number of alterations, scored as decreases and increases of band intensity with 2 arbitrary primers, were used as an estimation of the genomic damage fraction in 74 primary gastric cancers. Increases in DNA copy number were also analyzed by array comparative genomic hybridization in a subset of 30 cases. RESULTS: The number of altered bands varied among the tumors from none or a few to more than one third of the approximately 40 fingerprint bands. The relative values of genomic damage were consistent with the quantitative chromosomal alterations observed by array comparative genomic hybridization. When the tumors were stratified into 2 groups-above or below the cutoff of 0.22 for average genomic damage fraction-genomic damage fraction was a valuable prognostic indicator regardless of microsatellite instability status. Multivariate Cox analysis showed that the genomic damage fraction was a prognostic indicator, as well as a stage indicator (P = 0.0189). Survival was significantly diminished in tumors with a genomic damage fraction >0.22 (P = 0.0009). Moreover, in the 46 curative cases, genomic damage fraction was the only independent factor for predicting survival (P = 0.0061). CONCLUSIONS: Our results indicate that the degree of genomic damage estimated by arbitrarily primed polymerase chain reaction fingerprinting is a useful prognostic indicator for gastric cancer.
  • Expression profiles of pancreatic cancer cell lines infected with antisense K-ras-expressing adenoviral vector.
    Shumpei Ohnami, Kazunori Aoki, Kimiko Yoshida, Sumiko Ohnami, Kazuteru Hatanaka, Koichi Suzuki, Hiroki Sasaki, Teruhiko Yoshida
    Biochemical and biophysical research communications 309 4 798 - 803 2003年10月 [査読有り][通常論文]
     
    The point mutations of the K-ras gene occur in as high as 70-90% of the cases with adenocarcinoma of the pancreas and apparently represent one of the key and early events in the carcinogenesis. However, the specific influence of the K-ras activation on global gene expression profiles in pancreatic cancer cells has not been elucidated. In this study, to promote elucidation of the K-ras-triggered molecular cascade(s) in pancreatic cancer, four pancreatic cancer cell lines with K-ras point mutations were infected with an adenovirus vector expressing an antisense K-ras RNA (AxCA-AS), and the change of gene expression was analyzed by oligonucleotide-based microarrays containing 12,626 genes. Among the genes showing more than 2-fold differences in the expression levels between the control- and antisense-K-ras-transduced cells, 7 genes were commonly up-regulated and 4 genes were commonly down-regulated in three or all of the four pancreatic cancer cell lines transduced with AxCA-AS. The altered gene expression levels observed by microarrays were confirmed by real-time RT-PCR methods. Then, the expression of the 4 down-regulated genes was examined in the untransduced surgical specimens of pancreatic cancer. The G-protein coupled receptor RE2 and phenylethanolamine N-methyltransferase had negligible expression levels in all pancreatic cancers, whereas the syntaxin 1A and p120 catenin isoform were significantly up-regulated in pancreatic cancers containing K-ras mutations compared with a pancreatic cancer with wild type K-ras gene. The transcriptional regulation of those genes may be a part of the molecular cascades triggered by K-ras activation leading to the development and/or progression of pancreatic cancer.
  • Downregulation and growth inhibitory effect of epithelial-type Krüppel-like transcription factor KLF4, but not KLF5, in bladder cancer.
    Shunsuke Ohnishi, Sumiko Ohnami, Friedrich Laub, Kazunori Aoki, Koichi Suzuki, Yae Kanai, Kazunori Haga, Masahiro Asaka, Francesco Ramirez, Teruhiko Yoshida
    Biochemical and biophysical research communications 308 2 251 - 6 2003年08月 [査読有り][通常論文]
     
    Krüppel-like factors (KLFs) are key transcriptional regulators of cell differentiation and proliferation. Among the KLF family, the expression of KLF4 (GKLF) and KLF5 (IKLF) is highly restricted in the epithelial cells of several organs such as the gut and skin, and it has been reported that these epithelial-type KLF genes may be involved in colon carcinogenesis. Recently we found that Klf4 and Klf5 genes were significantly expressed in the developmental bladder epithelium of mice as well. Therefore, in this report we studied the involvement of the KLF4 and KLF5 genes in bladder carcinogenesis. First, we analyzed the expression of KLF4 and KLF5 in a variety of human bladder cancer cell lines and surgical specimens by RNA blot and in situ hybridization analyses. Both genes were highly expressed in the normal bladder epithelium, whereas KLF4, but not KLF5, was frequently downregulated in bladder cancer cell lines and cancer tissues. We then transduced the KLF4 and KLF5 genes into the bladder cancer cell lines using adenoviral vectors to examine the biological activities of the genes on those cells. The transduction of KLF4, but not KLF5, suppressed cell growth and induced apoptosis. Our study suggests that inactivation of KLF4 is one of the frequent steps towards bladder carcinogenesis.
  • Adenovirus-mediated gene transfer of interferon alpha inhibits hepatitis C virus replication in hepatocytes.
    Koichi Suzuki, Kazunori Aoki, Shumpei Ohnami, Kimiko Yoshida, Teruhisa Kazui, Nobuyuki Kato, Kazuaki Inoue, Michinori Kohara, Teruhiko Yoshida
    Biochemical and biophysical research communications 307 4 814 - 9 2003年08月 [査読有り][通常論文]
     
    Recently we reported that on-site interferon (IFN)-alpha production in the liver using an adenovirus vector can achieve a substantial confinement of IFN-alpha in the target organ and can improve liver fibrosis in a rat liver cirrhosis model. However, the major therapeutic effect of IFN for hepatitis C virus (HCV)-associated liver diseases is its antiviral effect on HCV. As a prelude to the in vivo HCV infection experiment using a primate animal model, here we examined the antiviral effect of IFN-alpha gene transfer into HCV-positive hepatocytes in vitro. The non-neoplastic human hepatocyte cell line PH5CH8 was inoculated with HCV-positive serum. Successful in vitro HCV replication and thus the validity of this model was confirmed by a strong selection for HCV variants determined by sequence analysis of the hypervariable region and an increase of HCV RNA estimated by real time TaqMan RT-PCR. One day after the inoculation of HCV, PH5CH8 cells were infected with adenoviral vectors encoding human IFN-alpha cDNA. HCV completely disappeared 9 days after the adenoviral infection, which is linked to the increase of 2('),5(')-oligoadenylate synthetase activity, suggesting that IFN-alpha produced by gene transfer effectively inhibits HCV replication in hepatocytes. This study supports the development of IFN-alpha gene therapy for HCV-associated liver diseases.
  • Adenovirus-mediated gene transfer of interferon alpha improves dimethylnitrosamine-induced liver cirrhosis in rat model.
    K Suzuki, K Aoki, S Ohnami, K Yoshida, T Kazui, N Kato, K Inoue, M Kohara, T Yoshida
    Gene therapy 10 9 765 - 73 2003年05月 [査読有り][通常論文]
     
    Several lines of evidence suggest that interferon (IFN)-alpha is effective in suppression of liver cirrhosis (LC) as well as hepatitis C virus (HCV) infection, which is a major cause of LC in Japan. However, IFN-alpha often causes systemic toxicity such as flu-like symptoms, which precludes the IFN-alpha dose escalation required for clinical efficacy. Since IFN-alpha is rapidly degraded in the blood circulation, only a small amount of subcutaneously injected IFN-alpha protein can reach the target organ, the liver. It is expected that on-site IFN-alpha production in the liver overcomes the limitation of the conventional parenteral IFN-alpha administration. An adenovirus vector expressing the rat IFN-alpha gene (AxCA-rIFN) was injected intravenously into rats with dimethylnitrosamine-induced LC. While the subcutaneous IFN-alpha protein injection led to a transient elevation of the cytokine both in the liver and serum, the vector-mediated IFN-alpha gene transduction induced a significant amount of IFN-alpha detected in the liver but not in the serum. The injection of AxCA-rIFN prevented the progression of the rat LC, and improved the survival rate of the treated rats. Although no significant toxicity was noted in the animals, we showed that IFN-alpha gene expression in the liver can be efficiently downregulated by the Cre/loxP-mediated shut-off system, in case the IFN-alpha overdose becomes a problem. The study suggested for the first time the advantage and feasibility of IFN-alpha gene therapy for LC.
  • Low mutation incidence in polymorphic noncoding short mononucleotide repeats in gastrointestinal cancer of the microsatellite mutator phenotype pathway.
    Koichi Suzuki, Tomoko Dai, Ikuko Suzuki, Yuichi Dai, Kentaro Yamashita, Manuel Perucho
    Cancer research 62 7 1961 - 5 2002年04月 [査読有り][通常論文]
     
    Frameshifts in short mononucleotide tracts (SMT) in genes, such as TGFbetaRII and BAX, are common in gastrointestinal tumors of the microsatellite mutator phenotype (MMP). The significance of less common mutations has been recently challenged because frequencies as high as 50% were reported in some noncoding SMTs in MMP colon cancer cell lines (L. Zhang, et al., Cancer Res., 61: 3801-3805, 2001). We did not confirm these findings after examining >50 MMP gastrointestinal cancers for mutations in eight SMT loci with the highest reported frequencies. In three of these loci, no clonal mutations were detected, and they were infrequent (2.9-6.7%) in the other five. Length polymorphisms are frequent (25.7-43.9%) in one-half of these SMTs, suggesting an explanation for the discrepancy. Because of the peculiar features of MMP tumors, low prevalence of mutations in cancer genes may not be a disqualifying criterion for their functionality.
  • Drug-induced apoptosis and p53, BCL-2 and BAX expression in breast cancer tissues in vivo and in fibroblast cells in vitro.
    K Suzuki, T Kazui, M Yoshida, T Uno, T Kobayashi, T Kimura, T Yoshida, H Sugimura
    Japanese journal of clinical oncology 29 7 323 - 31 1999年07月 [査読有り][通常論文]
     
    BACKGROUND: Chemotherapeutic management of breast cancers is a difficult task as they show significant differences in chemosensitivity. The present study was undertaken to determine the usefulness of the apoptosis-related factors as indicators of tumor sensitivity to 5'-deoxyfluorouridine (5'-DFUR) in breast cancers. METHODS: (1) Forty-six breast cancer patients were randomly assigned to a group in which oral 5'-DFUR (1200 mg/day) was administered for more than 5 days before operation (24 patients) and a control group who received no preoperative chemotherapy (22 patients). Surgical specimens were examined for the frequency of apoptotic cells [apoptotic index (AI)] by a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling method and for the expression of p53, BCL-2 and BAX by immunohistochemical staining. (2) Normal human diploid fetal lung fibroblast, IMR90 and SV40 transformed IMR90 were exposed to 5-FU. Apoptotic cells were detected by flow cytometry and BCL-2 and BAX mRNAs by real-time quantitative RT-PCR analysis. RESULTS: (1) No significant difference in the AIs or in BCL-2 and BAX scores was observed between the 5'-DFUR-treated and control groups. However, in the p53 negative subgroup (n = 36), AI and BAX scores were higher and BCL-2 scores lower in the 5'-DFUR group than in the control group (P = 0.006, 0.008 and 0.050, respectively). (2) The sensitivity of IMR90 was significantly decreased by SV40 transformation and the 5-FU-induced cytotoxicity was mainly due to induction of apoptosis. The BCL-2/BAX mRNA ratio was decreased in response to 5-FU in IMR90. These results correlated with our clinical data. CONCLUSIONS: Preoperative treatment with 5'-DFUR induced apoptosis and changes in BCL-2 and BAX expression in p53 negative breast cancers. p53 status, AI and the BCL-2/BAX ratio may be useful information for the choice of postoperative chemotherapy for breast cancer.
  • Pharmacokinetic advantages of a newly developed tacrolimus oil-in-water-type emulsion via the enteral route.
    T Uno, T Kazui, Y Suzuki, H Hashimoto, K Suzuki, B A Muhammad
    Lipids 34 3 249 - 54 1999年03月 [査読有り][通常論文]
     
    We developed an oleic acid oil-in-water (o/w)-type emulsion of a new tacrolimus formulation that presented an improvement in the delivery of the drug for oral absorption. This investigation was undertaken to assess a sustained release drug delivery system and selective drug transfer into the lymphatic system. The whole blood concentration profiles after oral administration at a dose of 2 mg/kg and bone marrow, spleen, liver, lung, small intestine, kidney, brain, and whole blood distribution after oral administration at a dose of 1 mg/kg of o/w emulsion formulation of tacrolimus (O/W group) were compared with those of commercially available formulation (T group) in the rat. The mean diameter of the o/w emulsion droplets was 0.47 microm immediately after preparation. The tacrolimus entrapping efficiency of o/w emulsion was 71.3+/-5.0% in 12 h and did not change for 2 d. The area under the whole blood concentration-time curve (AUC) in the O/W group was significantly higher (P<0.01) than that in the T group. In contrast, the values of constant elimination rate and total clearance in the O/W group were significantly lower (P<0.01) than those in the T group, with a comparative bioavailability of 115.9%. The tissue concentration of tacrolimus in the O/W group was significantly higher levels in the bone marrow, spleen, liver, lung, and small intestine, and significantly lower in the brain and kidney, relative to the T group. The o/w emulsion of tacrolimus may be an improved dosage form via the enteral route.

MISC

  • 癌免疫カンファレンスルーム(TOPICS 12) microRNAと大腸癌
    鈴木 浩一, 力山 敏樹 消化器外科 42 (8) 1237 -1240 2019年07月 [査読無し][通常論文]
  • Yuji Takayama, Koichi Suzuki, Kosuke Ichida, Taro Fukui, Fumiaki Watanabe, Nao Kakizawa, Yuta Muto, Rina Kikugawa, Fumi Hasegawa, Shingo Tsujinaka, Hiroshi Noda, Yasuyuki Miyakura, Fumio Konishi, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 35 2017年05月 [査読無し][通常論文]
  • Fumiaki Watanabe, Koichi Suzuki, Kosuke Ichida, Yuji Takayama, Taro Fukui, Nao Kakizawa, Yuhei Endo, Yuji Kaneda, Hiroshi Noda, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 35 2017年05月 [査読無し][通常論文]
  • Kosuke Ichida, Koichi Suzuki, Nao Kakizawa, Taro Fukui, Yuji Takayama, Fumiaki Watanabe, Takaharu Kato, Masaaki Saito, Kazushige Futsuhara, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 35 2017年05月 [査読無し][通常論文]
  • Nao Kakizawa, Koichi Suzuki, Ayuha Yoshizawa, Kosuke Ichida, Masaaki Saito, Kazushige Futsuhara, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 35 2017年05月 [査読無し][通常論文]
  • Takaharu Kato, Sergio Alonso, Yuta Muto, Hiroshi Noda, Yasuyuki Miyakura, Koichi Suzuki, Shingo Tsujinaka, Masaaki Saito, Taro Fukui, Kosuke Ichida, Yuji Takayama, Fumiaki Watanabe, Nao Kakizawa, Manuel Perucho, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 34 (15) 2016年05月 [査読無し][通常論文]
  • Practical applications of "liquid biopsy" in the colorectal cancer treatment.
    Koichi Suzuki, Yuji Takayama, Kosuke Ichida, Taro Fukui, Nao Kakizawa, Tamotsu Obitsu, Yuta Muto, Fumi Hasegawa, Fumiaki Watanabe, Rina Kikugawa, Takaharu Kato, Yuji Kaneda, Masaaki Saito, Shingo Tsujinaka, Kazushige Futsuhara, Yasuyuki Miyakura, Hiroshi Noda, Hirokazu Kiyozaki, Fumio Konishi, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 34 (15) 2016年05月 [査読無し][通常論文]
  • Clinical and molecular assessment of response to regorafenib.
    Nao Kakizawa, Koichi Suzuki, Yuji Takayama, Kosuke Ichida, Taro Fukui, Fumiaki Watanabe JOURNAL OF CLINICAL ONCOLOGY 34 (4) 2016年02月 [査読無し][通常論文]
  • Liquid biopsy leads to a paradigm shift in cancer treatment.
    Koichi Suzuki, Yuji Takayama, Kosuke Ichida, Taro Fukui, Nao Kakizawa, Yuta Muto, Fumiaki Watanabe, Fumi Hasegawa, Shingo Tsujinaka, Yuji Kaneda, Yasuyuki Miyakura, Hiroshi Noda, Hirokazu Kiyozaki, Fumio Konishi, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 34 (4) 2016年02月 [査読無し][通常論文]
  • 鈴木 浩一, 力山 敏樹 医学のあゆみ 255 (6) 661 -665 2015年11月 [査読無し][通常論文]
     
    近年のゲノムワイド関連解析(GWAS)は、Crohn病や潰瘍性大腸炎といった炎症性腸疾患(IBD)に関与する遺伝的要因の解明に大きな進歩をもたらした。最近の大規模国際共同研究により200を超えるIBDの疾患関連遺伝子座が同定され、疾患の発症にかかわるゲノム領域のほとんどが人種間で共通していることが明らかとなった。一方、遺伝的背景が同一である一卵性双生児の潰瘍性大腸炎、Crohn病の検討結果から疾患の発生率はそれぞれ16%、35%程度であり、遺伝的要因だけでIBDの病態を説明することが難しいこともわかってきた。遺伝的要因に加えて環境要因がかかわる相互作用の解明が必要となった。本稿では、潰瘍性大腸炎の遺伝的要因に関する研究の進展を確認し、遺伝的要因と環境を介在するエピゲノムの役割について最近の動向を概説する。(著者抄録)
  • Detection of satellite alpha transcript in sera, as a surrogate marker for the risk of development of multiple cancers in colorectal cancer patients.
    Kosuke Ichida, Koichi Suzuki, Yuji Takayama, Yuta Muto, Taro Fukui, Takaharu Kato, Masaaki Saito, Fumiaki Watanabe, Nao Kakizawa, Hirofumi Imoto, Hiroyuki Tanaka, Yuji Kaneda, Yusuke Taniyama, Shingo Tsujinaka, Osamu Takata, Yasuyuki Miyakura, Hiroshi Noda, Hirokazu Kiyozaki, Fumio Konishi, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 33 (15) 2015年05月 [査読無し][通常論文]
  • Emergence of KRAS mutation in detection of circulating tumor DNA during treatments for metastatic gastrointestinal cancer patients.
    Yuji Takayama, Koichi Suzuki, Tsutomu Daito, Kosuke Ichida, Taro Fukui, Yuta Muto, Nao Kakizawa, Hirofumi Imoto, Yusuke Taniyama, Yuji Kaneda, Hiroyuki Tanaka, Fumiaki Watanabe, Takaharu Kato, Fumi Hasegawa, Masaaki Saito, Shingo Tsujinaka, Yasuyuki Miyakura, Hiroshi Noda, Fumio Konishi, Toshiki Rikiyama JOURNAL OF CLINICAL ONCOLOGY 33 (15) 2015年05月 [査読無し][通常論文]
  • Johanna K. Samuelsson, Sergio Alonso, Koichi Suzuki, Manuel Perucho CANCER RESEARCH 71 2011年04月 [査読無し][通常論文]
  • 鈴木 浩一, 小西 文雄 Pharma Medica 27 (11) 29 -34 2009年11月 [査読無し][通常論文]
  • 鈴木 浩一, 岡田 晋一郎, 小西 文雄 Surgery Frontier 15 (3) 299 -303 2008年09月 [査読無し][通常論文]
  • Aging, DNA Hypomethylation, Genetic Alterations, and Gastrointestinal Cancer
    Manuel Perucho, Johanna Samuelsson, Andreas Leodolter, Sergio Alonso, Tatiana Ruiz-Larroya, Pepita Gimenez-Bonafe, Koichi Suzuki TUMOR BIOLOGY 29 45 -45 2008年 [査読無し][通常論文]
  • 消化器看護 患者個別に対応する!術後ケアのポイント 食道癌切除後のケア
    鈴木 浩一, 小林 利彦, 和田 英俊, 渡辺 浩, 小西 由樹子 消化器・がん・内視鏡ケア 11 (4) 025 -032 2006年10月 [査読無し][通常論文]
  • 鈴木 浩一, 坂元 吾偉, 秋山 太, 霞 富士雄 日本乳癌検診学会誌 = Journal of Japan Association of Breast Cancer Screening 6 (3) 323 -327 1997年10月 [査読無し][通常論文]
  • 坂元 吾偉, 秋山 太, 鈴木 浩一 日本乳癌検診学会誌 = Journal of Japan Association of Breast Cancer Screening 5 (1) 41 -46 1996年04月 [査読無し][通常論文]

共同研究・競争的資金等の研究課題

  • 染色体不安定性を標的とした診断、治療アプローチの確立
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2020年04月 -2023年03月 
    代表者 : 鈴木 浩一
  • 染色体不安定性が関わる多発癌発生の機序の解明
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2020年04月 -2023年03月 
    代表者 : 野田 弘志, 鈴木浩一, 力山敏樹
  • 食道胃接合部癌発癌におけるゲノムワイドなエピジェネティック異常の網羅的解析
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2020年04月 -2023年03月 
    代表者 : 斉藤 正昭, 鈴木浩一, 力山敏樹
  • 染色体不安定性が関わる多発癌発生の機序の解明
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2017年04月 -2020年03月 
    代表者 : 野田 弘志, 鈴木 浩一, 力山 敏樹
     
    【研究目的】癌患者は、術後の残存臓器に新たな腫瘍性病変を高率に生じます。その機序は明らかではありませんが、発癌の母地「Field Cancerization; FC」の関与が考えられます。FCにはさまざまな遺伝子の異常が報告されていますが、我々はメチル化修飾に注目してきました。正常な染色体分配にはセントロメア領域のメチル化が必須で、そこから転写されるnon-cording RNAであるSatelliteαtranscript(SatA)により染色体の均等分配が司られます。我々はセントロメア領域のメチル化異常によりSatAが過剰発現し、特定の染色体に数的異常が生じる事を明らかにしました。この変化は癌部のみならず癌の背景粘膜でも認められ、特に多発癌の背景粘膜で高値を示すことから、SatAは多発癌のFCに関わると考えています。本研究ではSatAが関わる多発癌発生の機序を明らかにします。 【研究実績】レンチウイルスを用いて乳腺上皮細胞株にSatAを遺伝子導入しました。染色体不安定性をもたらす有糸分裂異常を免疫細胞化学によって調べたところ、Abnormal segregation、MicronucleiおよびAnaphase bridgeといった有糸分裂異常の頻度はSatAの過剰発現細胞において有意に増加していることを明らかにしました(掲載論文3)。マウスMajarSatA配列を搭載したレンチウイルスベクターを作成し、マウスの乳癌発生モデルを用いて行いました。乳癌発生を促進するためDMBAを経口投与し、マウス生体の乳管内にレンチウイルスを投与したところ、観察期間内では介入群および対象群とも肉眼的には乳癌の発生は見られませんでした。
  • 血中遊離DNAのモニタリングによる疾患プロファイルの構築と個別化治療戦略
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2017年04月 -2020年03月 
    代表者 : 宮倉 安幸, 鈴木 浩一, 力山 敏樹
     
    【研究目的】我々は腫瘍のゲノムプロファイルがその進展・転移、治療の過程で変化する事に注目し、ゲノムプロファイルのモニタリングとその情報に基づく癌治療戦略の構築を進めてきました。しかし頻繁な生検や転移巣の検体採取が必要で、煩雑性や過大侵襲が大きな問題でした。その問題を解決したのが、血中に遊離した希少な腫瘍細胞やDNAを検出する技術「Liquid biopsy」です。我々が注目したのはKRAS遺伝子で、切除不能大腸癌の治療標的分子です。本研究では、薬剤耐性のみならず感受性回復の指標としての意義を検証し、薬剤の再導入の可能性を探ります。また原発巣と転移巣のKRAS statusの解離に注目し、KRAS statusに基づく治療介入方法を検討します。新たな医療アプローチとして、臨床応用を目指します。 【研究実績】平成30年度は、 (1)KRAS遺伝子の血中モニタリングの継続:「Liquid biopsy」でKRAS変異を血中モニタリングする事によって薬剤耐性の獲得をより鋭敏に捉える事が可能となり、薬剤耐性のみならず感受性回復、薬剤の再導入のへの指標となる事を報告しました。(Takayama Y.Oncotarget.2018) (2)液滴ソーティングによる塩基配列解析と簡易解析システムのアプリケーション化:標的分子としてEGFRのモニタリングを追加しました。株式会社オンチップ・バイオテクノロジーズのソーティングシステムを用いて循環腫瘍細胞の採取を行い、循環腫瘍DNAとの比較を行いました。大腸癌の分子標的治療薬であるRegorafenibの投与前後で循環腫瘍DNAを採取し、EGFRの発現レベルを比較すると、Regorafenibを投与することによりEGFRの発現レベルが上昇する事を確認しました。抗EGFR抗体の再投与が有効性を考える上で意義のある結果です。
  • ゲノムダメージに関わる標的分子の特定と血中モニタリングシステムの開発
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2016年04月 -2020年03月 
    代表者 : 鈴木 浩一, 力山 敏樹
     
    【研究目的】年齢と共に増加する発癌リスクの要因としてゲノム損傷の蓄積が考えられます。その契機は遺伝子複製の際の染色体分配です。正常な染色体分配には染色体のセントロメア領域のメチル化が必須です。本研究では、加齢に伴う染色体のメチル化異常に着目し、SatAを介する染色体不安定性の標的染色体からゲノム損傷に関わる標的分子を特定します。それを血中モニタリングすることで発癌リスクを経時的に評価するシステムを構築します。 【研究実績】「Satelliteαtranscript」を介するゲノムのダメージのメカニズムの検証:レンチウイルスを用いて乳腺上皮細胞株にSatAを遺伝子導入しました。染色体不安定性をもたらす有糸分裂異常を免疫細胞化学によって調べたところ、Abnormal segregation、MicronucleiおよびAnaphase bridgeといった有糸分裂異常の頻度はSatAの過剰発現細胞において有意に増加していることを明らかにしました(掲載論文3)。SatAが両側乳癌や他臓器癌合併につながるfield cancerizationに関与するかどうかを調べるために、乳癌患者167人の解析を行いました。その結果、非癌部におけるSatA高発現症例は、低発現症例に比較して22倍の両側乳癌のリスクを持ち、さらに11倍の他臓器癌合併のリスクを持つという結果を導き出すことができました。現在Oncology reportに投稿中です。 微量遺伝子の血中検出と塩基配列解析システムの確立:患者の血漿検体を収集し、デジタルPCRを用いてKRAS血中モニタリングを行ってきました。大腸がん患者(掲載論文1)及び膵がん患者において、治療効果や予後因子としての有用性が示されました(Clinical Cancer Research 投稿中)。
  • DNAメチル化プロファイルによる癌易罹患性の予測
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2016年04月 -2020年03月 
    代表者 : 力山 敏樹, 鈴木 浩一
     
    【研究目的】本研究は、正常組織で観察される遺伝子修飾異常(DNAメチル化異常)の情報をもとに、癌のかかりやすさを予測しようとするものです。DNAメチル化異常は癌組織のみならず、非癌部組織でも認められるため、その情報を癌発症の予測や早期診断に利用します。具体的には、癌組織で異常メチル化のため遺伝子発現が低下している遺伝子を同定し、それらの遺伝子の異常メチル化の程度を背景粘膜でも検出します。背景粘膜の異常メチル化の程度と遺伝子発現の相関を解析し、異常メチル化の進行に伴い遺伝子の発現が低下していく過程を捉えられれば、それは発癌過程の段階的変化を表現していると考えられます。我々が独自に開発したDNAメチル化マイクロアレイ、発現アレイを用いる事により、網羅的に遺伝子異常、遺伝子修飾異常を捉えることが可能となり、分子生物学的な発癌プロファイルの作成が可能となります。それは発癌リスクのバイオマーカーとして有用性が期待されます。 【研究実績】平成30年度は、 発現アレイで遺伝子発現の程度を評価し、Pyro Markでメチル化シトシン定量し、アレイの結果の検証を行います。そしてクラスター解析、DAVID解析で関連遺伝子を同定し、メチル化プロファイルを作成します。そしてDNAメチル化異常とその遺伝子の発現の情報を統合させ、分子生物学的な発癌リスクのプロファイルの作成を目標とします。 我々が独自に開発したDNAメチル化マイクロアレイ、MS-AFLP法を用いて癌部と非癌部で同じ変化を示す遺伝子断片を同網羅的に検索し、クラスター解析、DAVID解析を行ったところ、WNTsignal、AXON guidanceおよびHomeoboxに関わる遺伝子群を同定しました。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2016年04月 -2019年03月 
    代表者 : 斉藤 正昭, 鈴木 浩一, 力山 敏樹
     
    我々は、DNAメチル化異常によりセントロメア領域の反復配列であるSatAが過剰発現し、特定の染色体を標的として数的異常が生じる事を明らかにした。この変化は特に多発癌の背景粘膜で高値を示すことから、SatAは潜在的な癌の発生母地(field defect)を反映すると考えられる。本研究ではSatAのDNA脱メチル化異常に注目し、胃癌の多発発生への関与を検討した。SatAの脱メチル化レベルは、単発胃癌に比べ多発胃癌と有意に高く(P<0.001)、多発胃癌の存在を予測する独立因子であった(P = 0.008)。SatA はfield defectを介して胃がんの多発発生に関わっていると考えられた。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2015年04月 -2018年03月 
    代表者 : 辻仲 眞康, 鈴木 浩一, 力山 敏樹
     
    上皮細胞が間葉系様細胞に変化する現象、上皮間葉系移行が癌の転移のメカニズムとして注目されていますが、ヒト臨床検体では証明されていません。その理由として腫瘍内不均一性に起因する遺伝子発現の偏在が考えられます。そこで、臨床検体を用いて癌先進部の遺伝子発現を中心部の発現で補正した遺伝子発現比を算出し、転移の有無で比較したところ、転移に関わる473遺伝子を同定し、adhesion、WntやVEGF等の上皮間葉系移行を促進する伝達経路を捉えました。すなわち転移を有する癌では、これらのシグナルが先進部で広く活性化し、その中でもVEGFAが高発現している約20%の大腸癌は極めて高い再発・転移を示しました。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2014年04月 -2017年03月 
    代表者 : 小西 文雄, 鈴木 浩一
     
    近年、鋸歯状ポリープという鋸歯状構造を有するポリープを総称する概念が提唱された。その中でも鋸歯状腺腫(SSA)はマイクロサテライト不安定性(MSI)大腸癌の前癌病変として注目されるようになった。我々は、SSAとMSI大腸癌に共通するメチル化プロファイルに注目し、独自に開発したDNAメチル化アレイを用いて127関連遺伝子を特定した。その1つであるAXINは大腸癌抑制遺伝子であるAPCとともに複合体を形成しβカテニンの分解に携わる。AXINのメチル化の程度は腺腫から癌へ進行すると共に上昇し、それに伴いその遺伝子発現は低下する。AXINは鋸歯状ポリープを介した発癌過程に深く関わっていると考えられる。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2013年04月 -2016年03月 
    代表者 : 鈴木 浩一, 力山 敏樹
     
    年齢と共に増加する発癌リスクの要因としてゲノム損傷の蓄積が考えられます。その契機は遺伝子複製の際の染色体分配です。正常な染色体分配には染色体のセントロメア領域のメチル化が必須です。セントロメア領域は反復配列であるSatelliteα配列より構成され、そこから転写されるnon-cording RNA であるSatelliteαtranscript(SatA)により染色体の均等分配が司られます。我々は、セントロメア領域のメチル化異常によりSatA が過剰発現し、特定の染色体を標的として数的異常が生じる事を明らかにしました。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2013年04月 -2016年03月 
    代表者 : 力山 敏樹, 鈴木 浩一
     
    DNAメチル化異常は癌組織のみならず非癌部組織でも認められます。そこで非癌部組織で観察されるDNAメチル化異常の情報をもとに、癌のかかりやすさを予測しようと考えました。癌組織で異常メチル化のため発現が低下している遺伝子を同定し、その異常メチル化の程度を背景粘膜で解析しました。潰瘍性大腸炎では、発癌に関連するDNAメチル化異常を非癌部組織で捉える事により、癌の易罹患性を予測する有用な指標となる事を示しました。さらに大腸癌ではDNAメチル化マイクロアレイを用いて、その発癌経路に関わるメチル化異常の違いを評価し、一連の発癌経路に関わる遺伝子の変化が癌と腺腫で共有されている事を明らかにしました。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2009年 -2011年 
    代表者 : 鈴木 浩一
     
    本研究は、正常組織で観察される遺伝子修飾異常(DNAメチル化異常)の情報をもとに、癌のかかりやすさを予測しようとするものです。胃癌、大腸癌の初回手術の非癌部背景粘膜のDNA脱メチル化異常に着目し、多発癌の発生リスクのバイオマーカーとしての有用性を示しました。また、我々が独自に開発したDNAメチル化マイクロアレイを用いる事により、網羅的に遺伝子異常、遺伝子修飾異常を捉えることが可能となり、分子生物学的な疾患プロファイルの作成が期待されます。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2007年 -2008年 
    代表者 : 鈴木 浩一, 宮木 祐一郎, 小泉 圭, 岡田 晋一郎
     
    DNAメチル化異常は癌のみならず、癌発症の背景となる慢性胃炎や潰瘍性大腸炎等の非癌部組織にも認められる。本研究では我々が独自に開発したDNAメチル化マイクロアレイを用いて、DNAメチル化異常をゲノム全域に渡り網羅的に捕捉し、担癌、非担癌潰瘍性大腸炎の非癌部組織にみられる変化を癌部組織と比較検討することにより、潰瘍性大腸炎の発癌に関わる遺伝子群(メチル化異常138カ所、脱メチル化異常221カ所)を同定した。
  • 日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2006年 -2008年 
    代表者 : 常吉 俊宏, 鈴木 浩一
     
    ヒト正常皮膚繊維芽細胞(NHDF)とピロリ菌(HP)を混合培養し、ヒトDNAのメチル化(エピジェネティック変異)を検出したところ、HP混合でNHDFのメチル化(最大44%)・脱メチル化(同8%)が起こった。緑茶カテキンを混合培養系に添加すると脱メチル化方向に変化した。マイクロアレイを用いたところヒトのDNAメチル化酵素、ヒストン脱アセチル化酵素など重要遺伝子がHP混合でメチル化されることがわかった。
  • 消化器癌におけるDNAメチル化制御の異常とゲノムの不安定性に関する検討
    日本学術振興会:科学研究費助成事業 基盤研究(C)
    研究期間 : 2005年 -2006年 
    代表者 : 鈴木 浩一
     
    癌の発生、進展には遺伝子配列の変化を伴う遺伝子異常と、DNAメチル化のような遺伝子配列の変化を伴わない遺伝子修飾の異常が共に関与している。プロモーター領域のメチル化は癌抑制遺伝子の転写活性を失活させるが、片方のアレルの遺伝子異常と共にKnudsonのsecond hitとして他方のアレルに働く。一方脱メチル化はゲノム全域にわたり認められ、ゲノムの不安定性を惹起する事が示唆されている。このように遺伝子異常と遺伝子修飾の異常は互いに密接に影響しあい、癌化に深く関わっていると考えられ、癌の病体解明には相互の役割を明らかにすることが肝要である。そこで我々は遺伝子修飾の異常をゲノム全域に渡り網羅的に捕捉、評価し、遺伝子異常と関連づけて検討した。 (対象と方法)胃癌86症例、大腸癌67症例を対象とした。遺伝子修飾の異常は、約150のCpG lociのDNAメチル化の変化をmethylation sensitive amplified fragment length polymorphism(MS-AFLP)法を用いて検出した。遺伝子異常は、約100の染色体領域のDNAコピー数の変化をarbitrarily primed PCR(AP-PCR)法を用いて検出した。 (結果)遺伝子修飾の異常、遺伝子異常の分布は共になだらかな増加曲線を描いた。遺伝子修飾異常はメチル化、脱メチル化共に加齢に伴い増加したが、遺伝子異常は年齢との相関を認めなかった。メチル化異常は脱メチル化異常と相関した。遺伝子修飾異常はメチル化、脱メチル化異常共に遺伝子異常と相関したが、特に脱メチル化異常がより有為に遺伝子異常に関与していた。 (結語)遺伝子修飾の異常、特に脱メチル化異常は加齢に伴いその頻度を増し、ゲノムの不安定性を惹起することにより遺伝子異常を引き起こし癌化に関わっていることが示唆された。


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