Researchers Database

suzuki kouichi

    Associate Professor
Last Updated :2021/12/05

Researcher Information

Research funding number

  • 70332369

J-Global ID

Research Areas

  • Life sciences / Digestive surgery

Published Papers

  • 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
    Scientific reports 11 (1) 20797 - 20797 2021/10 [Refereed]
    Despite the acceptance of carbohydrate antigen 19-9 (CA19-9) as a valuable predictor for the prognosis of pancreatic ductal adenocarcinoma (PDAC), its cutoff value remains controversial. Our previous study showed a significant correlation between CA19-9 levels and the presence of KRAS-mutated ctDNA in the blood of patients with PDAC. Based on this correlation, we investigated the optimal cutoff value of CA19-9 before surgery. Continuous CA19-9 values and KRAS-mutated ctDNAs were monitored in 22 patients with unresectable PDAC who underwent chemotherapy between 2015 and 2017. Receiver operating characteristic curve analysis identified 949.7 U/mL of CA19-9 as the cutoff value corresponding to the presence of KRAS-mutated ctDNA. The median value of CA19-9 was 221.1 U/mL. Subsequently, these values were verified for their prognostic values of recurrence-free survival (RFS) and overall survival (OS) in 60 patients who underwent surgery between 2005 and 2013. Multivariate analysis revealed that 949.7 U/mL of CA19-9 was an independent risk factor for OS and RFS in these patients (P = 0.001 and P = 0.010, respectively), along with lymph node metastasis (P = 0.008 and P = 0.017), unlike the median CA19-9 level (P = 0.150 and P = 0.210). The optimal CA19-9 level contributes to the prediction of prognosis in patients with PDAC before surgery.
  • 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 2020/05 [Refereed][Not invited]
    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 2019/10 [Refereed][Not invited]
    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 2019/08 [Refereed][Not invited]
    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 2019/05 [Refereed][Not invited]
    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 [Refereed][Not invited]
    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 [Refereed][Not invited]
    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 2018/05 [Refereed][Not invited]
    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 2018/05 [Refereed][Not invited]
    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 2018/04 [Refereed][Not invited]
    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 [Refereed][Not invited]
    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 [Refereed][Not invited]
    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 2017/10 [Refereed][Not invited]
    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 2017/09 [Refereed][Not invited]
    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 2017/08 [Refereed][Not invited]
    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 2017/06 [Refereed][Not invited]
    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 2017/05 [Refereed][Not invited]
    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 2017/04 [Refereed][Not invited]
    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.
  • 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 2016/10 [Refereed][Not invited]
    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 2016/09 [Refereed][Not invited]
    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 2015/02 [Refereed][Not invited]
    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 2015/01 [Refereed][Not invited]
    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 [Refereed][Not invited]
    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 2014/12 [Refereed][Not invited]
    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.
  • 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 2049-9450 2014/09 [Refereed][Not invited]
    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 2014/06 [Refereed][Not invited]
    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 2012/11 [Refereed][Not invited]
    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 2012/06 [Refereed][Not invited]
    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 2012/04 [Refereed][Not invited]
    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 2012/04 [Refereed][Not invited]
    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 2012/03 [Refereed][Not invited]
    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 2012/01 [Refereed][Not invited]
    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.
  • Takafumi Maeda, Koichi Suzuki, Kazutomo Togashi, Mitsuhiro Nokubi, Masaaki Saito, Shingo Tsujinaka, Hidenori Kamiyama, Fumio Konishi
    Experimental and therapeutic medicine 2 (4) 695 - 700 1792-0981 2011/07 [Refereed][Not invited]
    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.
  • Johanna K. Samuelsson, Sergio Alonso, Koichi Suzuki, Manuel Perucho
    CANCER RESEARCH 71 0008-5472 2011/04 [Not refereed][Not invited]
  • Kok-Yang Tan, Fumio Konishi, Koichi Suzuki
    Surgery today 40 (4) 385 - 7 2010/04 [Refereed][Not invited]
    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 2009/07 [Refereed][Not invited]
    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 2008/04 [Refereed][Not invited]
    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.
  • Manuel Perucho, Johanna Samuelsson, Andreas Leodolter, Sergio Alonso, Tatiana Ruiz-Larroya, Pepita Gimenez-Bonafe, Koichi Suzuki
    TUMOR BIOLOGY 29 45 - 45 1010-4283 2008 [Not refereed][Not invited]
  • 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 2007/05 [Refereed][Not invited]
    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.
  • Koichi Suzuki, Ikuko Suzuki, Andreas Leodolter, Sergio Alonso, Shina Horiuchi, Kentaro Yamashita, Manuel Perucho
    Cancer cell 9 (3) 199 - 207 1535-6108 2006/03 [Refereed][Not invited]
    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.
  • 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 1078-0432 2005/08 [Refereed][Not invited]
    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.
  • 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 1099-498X 2004/10 [Refereed][Not invited]
    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.
  • 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 0016-5085 2003/11 [Refereed][Not invited]
    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.
  • 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 0006-291X 2003/10 [Refereed][Not invited]
    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.
  • 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 0006-291X 2003/08 [Refereed][Not invited]
    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.
  • 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 0006-291X 2003/08 [Refereed][Not invited]
    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.
  • K Suzuki, K Aoki, S Ohnami, K Yoshida, T Kazui, N Kato, K Inoue, M Kohara, T Yoshida
    Gene therapy 10 (9) 765 - 73 0969-7128 2003/05 [Refereed][Not invited]
    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.
  • Koichi Suzuki, Tomoko Dai, Ikuko Suzuki, Yuichi Dai, Kentaro Yamashita, Manuel Perucho
    Cancer research 62 (7) 1961 - 5 0008-5472 2002/04 [Refereed][Not invited]
    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.
  • 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 0368-2811 1999/07 [Refereed][Not invited]
    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.
  • T Uno, T Kazui, Y Suzuki, H Hashimoto, K Suzuki, B A Muhammad
    Lipids 34 (3) 249 - 54 0024-4201 1999/03 [Refereed][Not invited]
    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.

Conference Activities & Talks

  • 大腸癌肝転移に対する治療戦略 大腸癌肝転移に対する外科的治療選択における形態学的変化の有用性の検証
    福井 太郎, 鈴木 浩一, 市田 晃佑, 柿澤 奈緒, 武藤 雄太, 渡部 文昭, 兼田 裕司, 宮倉 安幸, 野田 弘志, 力山 敏樹
    日本消化器外科学会総会  2017/07  (一社)日本消化器外科学会
  • 膵腫瘍性病変における組織と血中のKrasモニタリングの臨床的意義
    渡部 文昭, 鈴木 浩一, 遠藤 裕平, 市田 晃佑, 福井 太郎, 柿澤 奈緒, 兼田 裕司, 野田 弘志, 力山 敏樹
    日本外科学会定期学術集会抄録集  2017/04  (一社)日本外科学会
  • 切除不能胃癌におけるConversion therapy可能症例の検討
    齊藤 正昭, 清崎 浩一, 染谷 崇徳, 石岡 大輔, 鈴木 浩一, 兼田 裕司, 辻仲 眞康, 宮倉 安幸, 野田 弘志, 力山 敏樹
    日本消化器外科学会総会  2016/07  (一社)日本消化器外科学会
  • 膵腫瘍性病変におけるkrasモニタリングの臨床的意義
    渡部 文昭, 鈴木 浩一, 野田 弘志, 市田 晃佑, 高山 裕司, 福井 太郎, 柿澤 奈緒, 武藤 雄太, 兼田 裕司, 力山 敏樹
    日本消化器外科学会総会  2016/07  (一社)日本消化器外科学会
  • P0CY1胃癌症例に対する治療戦略
    石岡 大輔, 齊藤 正昭, 渡部 文昭, 兼田 裕司, 辻仲 眞康, 鈴木 浩一, 野田 弘志, 宮倉 安幸, 清崎 浩一, 力山 敏樹
    日本消化器外科学会総会  2016/07  (一社)日本消化器外科学会
  • 大腸癌同時性肝転移の初回治療戦略
    市田 晃佑, 野田 弘志, 鈴木 浩一, 柿澤 奈緒, 渡部 文昭, 兼田 裕司, 力山 敏樹
    日本肝胆膵外科学会・学術集会プログラム・抄録集  2016/06  (一社)日本肝胆膵外科学会
  • 2度のRFAで制御している膵癌肝転移の1例
    渡部 文昭, 野田 弘志, 柿澤 奈緒, 兼田 裕司, 齊藤 正昭, 辻仲 眞康, 鈴木 浩一, 宮倉 安幸, 清崎 浩一, 力山 敏樹
    日本肝胆膵外科学会・学術集会プログラム・抄録集  2016/06  (一社)日本肝胆膵外科学会
  • P0CY1胃癌症例に対する治療戦略
    石岡 大輔, 齊藤 正昭, 染谷 崇徳, 福田 臨太郎, 渡部 文昭, 兼田 裕司, 菊川 利奈, 辻仲 眞康, 野田 弘志, 宮倉 安幸, 鈴木 浩一, 清崎 浩一, 遠山 信幸, 力山 敏樹
    日本外科学会定期学術集会抄録集  2016/04  (一社)日本外科学会
  • 当院における上部消化管神経内分泌腫癌の検討 食道・胃
    高田 理, 石岡 大輔, 福田 臨太郎, 小櫃 保, 柿澤 奈緒, 斎藤 正昭, 染谷 崇徳, 菊川 怜奈, 長谷川 芙美, 渡部 文昭, 兼田 裕司, 辻仲 眞康, 鈴木 浩一, 宮倉 安幸, 野田 弘志, 蛭田 昌宏, 土橋 洋, 野首 光弘, 山田 茂樹, 力山 敏樹
    日本外科学会定期学術集会抄録集  2016/04  (一社)日本外科学会
  • 形態学的応答の評価によるレスキュー肝切除術(Rescue liver resection by assessment of morphological response)
    福井 太郎, 鈴木 浩一, 市田 晃佑, 高山 裕司, 柿澤 奈緒, 武藤 雄太, 渡部 文昭, 兼田 裕司, 宮倉 安幸, 野田 弘志, 力山 敏樹
    日本外科学会定期学術集会抄録集  2016/04  (一社)日本外科学会
  • 武藤 雄太, 鈴木 浩一, 前田 孝文, 渡部 文昭, 加藤 高晴, 河村 裕, 小西 文雄, 力山 敏樹
    日本外科学会雑誌  2013/03  一般社団法人日本外科学会
  • 加藤 高晴, 鈴木 浩一, 神山 英範, 斎藤 正昭, 武藤 雄太, 小西 文雄, 力山 敏樹
    日本外科学会雑誌  2013/03  一般社団法人日本外科学会
  • 加藤 高晴, 鈴木 浩一, 岡田 晋一郎, 神山 英範, 前田 孝文, 齊藤 正昭, 小泉 圭, 宮木 祐一郎, 小西 文雄
    日本外科学会雑誌  2012/03  一般社団法人日本外科学会
  • 齊藤 正昭, 鈴木 浩一, 加藤 高晴, 前田 孝文, 千葉 文博, 高田 理, 清崎 浩二, 小西 文雄
    日本外科学会雑誌  2012/03  一般社団法人日本外科学会


Research Grants & Projects

  • Clinical application of the blood monitoring system in accordance with genomic information of circulating tumor cells
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2021/04 -2024/03 
    Author : 宮倉 安幸, 鈴木 浩一
  • 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2020/04 -2023/03 
    Author : 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2020/04 -2023/03 
    Author : 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2020/04 -2023/03 
    Author : 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2017/04 -2020/03 
    Author : 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2017/04 -2020/03 
    Author : 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2016/04 -2020/03 
    Author : 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
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2016/04 -2020/03 
    Author : 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。
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2016/04 -2019/03 
    Author : SAITO Masaaki
    We previously reported that overexpression of satellite alpha transcript (SAT) facilitates chromosomal instability, which is involved in the development of multiple tumors in patients with colorectal. The centromere consists of satellite alpha repetitive sequences, which are ideal targets for DNA hypomethylation, resulting in the overexpression of SAT. In this study, we elucidated the significance of DNA demethylation of SAT in the development of multiple tumors in patients with gastric cancer. Relative demethylation level of SAT was calculated from 103 gastric patients. Patients with multiple gastric cancer (n=20) showed higher demethylation levels than those in patients with single cancer (n=83, P<0.001). Additionally, multivariate analysis revealed that the demethylation level of SAT was a significant factor for predicting multiple gastric cancer (P=0.008). Our data indicated that DNA demethylation of SAT was involved in the development of multiple gastric cancer.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2015/04 -2018/03 
    Author : TSUJINAKA Shingo
    Increasing evidence suggests that epithelial-mesenchymal transition (EMT) plays an important role in tumor progression and metastasis formation but there is no evidence in human tumors. It may be the great diversity in cellular organization, known as intratumor heterogeneity. We calculated the expression ratios between the tumor center and invasive front in each tumor. This approach enabled recognition of the activation of the VEGF and Wnt signaling pathways, which were involved in metastasis via promotion of EMT. They were preferentially activated at the invasive front in metastatic tumors. Furthermore, we found 20 % of colorectal patients showed the overexpression of VEGF-A and they were more likely to relapse and metastasize.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2014/04 -2017/03 
    Author : Konishi Fumio
    A concept of sessile serrated polyp has been proposed as a series of polyps with serration. Recent works led to recognize sessile serrated adenoma as precursor to many of the sporadic colorectal cancers with microsatellite instability (MSI). We focused on the methylation profile sharing between SSA and MSI cancer, which resulted in identification of 127 associated genes by an array-based methylation sensitive amplified fragment length polymorphism (MS-AFLP) method. AXIN is one of those genes, which cooperates with the tumor suppressor gene, APC to depredate beta-catenin. Its levels of methylation alterations increased along with progression from adenoma to carcinoma, and its expression levels negatively correlated with methylation. These findings suggested that AXIN was involved in the serrated pathway originated from sessile serrated polyp.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2013/04 -2016/03 
    Author : Suzuki Koichi, RIKIYAMA Toshiki
    Impairment of stability in chromosomal structure facilitates abnormal segregation of chromosome, increasing the risk of carcinogenesis. Segregation is managed by appropriate methylation, leading to heterochromatin structure at centromere. Insufficient methylation such as demethylation fails to heterochromatin due to overexpression of satellite alpha transcript (SatA), which is the non-coding RNA transcribed from satellite alpha sequences at centromere. We showed SatA expression was increased in colorectal cancer, in which satellite sequences were demethylated, resulting in chromosomal instability. We verified this machinery in vitro. The lentiviral vector expressing human SatA was constructed and transduced in cultured cells. Copy number alterations were observed at several chromosomes in transduced cells but not in mock cells. Alterations were frequently seen at chromosome 8q and 20q, which is consistent with the results in clinical specimens.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2013/04 -2016/03 
    Author : Rikiyama Toshiki, SUZUKI Koichi
    In this study, we explore DNA methylation alterations in non-cancer tissues to evaluate the susceptibility of cancer. Methylation alterations are observed in both cancer and non-cancer tissues, so we elucidate the levels in non-cancer tissues to link with the predisposition of cancer or detection of early cancer. We identified methylation alterations in some genes decreased in cancer tissues, and explore the levels of methylation in these genes in non-cancer tissues. In ulcerative colitis, we showed that methylation alteration in non-cancer tissues could be a useful biomarker for the predisposition of colitic-cancer. Comprehensive analysis of DNA methylation array in colon cancer revealed that methylation alterations underlying colon cancer development are shared between cancer and adenomas.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2009 -2011 
    Author : SUZUKI Koichi
    We evaluated the levels of demethylation alterations in normal tissues from patients who had undergone gastrectomy or colectomy due to cancer development. Our data showed that high levels of demethylation alterations were frequently seen in patients who developed synchronous or metachronous double cancers as compare with those did not. Further, we developed a novel microarray platform based on methylation-sensitive amplified fragment length polymorphism DNA fingerprinting to perform the genome-wide severance of methylation alterations, which would enable us to make biological profiles according to the several diseases including cancer.
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2007 -2008 
    Author : SUZUKI Kouichi, MIYAKI Yuuichirou, KOIZUMI Kei, OKADA Shinichirou
  • Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2006 -2008 
    Author : TSUNEYOSHI Toshihiro, SUZUKI Koichi
  • DNA methylation alterations and their relationship to genomic instability in gastrointestinal cancer
    Japan Society for the Promotion of Science:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)
    Date (from‐to) : 2005 -2006 
    Author : SUZUKI Koichi
    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.

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