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Anoctamin 5 regulates the cell cycle and affects prognosis in gastric cancer

World Journal of Gastroenterology 2022年第32期28卷 4649-4667页

作者： Tomoyuki Fukami Atsushi Shiozaki Toshiyuki Kosuga Michihiro Kudou Hiroki Shimizu Takuma Ohashi Tomohiro Arita Hirotaka Konishi Shuhei Komatsu Takeshi Kubota Hitoshi Fujiwara Kazuma Okamoto Mitsuo Kishimoto Yukiko Morinaga Eiichi Konishi eigo otsuji Division of Digestive Surgery Department of SurgeryKyoto Prefectural University of MedicineKyoto 602-8566Japan Department of Pathology Kyoto City HospitalKyoto 604-8845Japan Department of Pathology Kyoto Prefectural University of MedicineKyoto 602-8566Japan

BACKGROUND Anoctamin 5(ANO5)/transmembrane protein 16E belongs to the ANO/transmembrane protein 16 anion channel family.ANOs comprise a family of plasma membrane proteins that mediate ion transport and phospholipid scrambling and regulate other membrane proteins in numerous cell types.Previous studies have elucidated the roles and mechanisms of ANO5 activation in various cancer types.However,it remains unclear whether ANO5 acts as a plasma membrane chloride channel,and its expression and functions in gastric cancer(GC)have not been investigated.AIM To examine the role of ANO5 in the regulation of tumor progression and clinicopathological significance of its expression in GC.METHODS Knockdown experiments using ANO5 small interfering RNA were conducted in human GC cell lines,and changes in cell proliferation,cell cycle progression,apoptosis,and cellular movement were assessed.The gene expression profiles of GC cells were investigated following ANO5 silencing by microarray analysis.Immunohistochemical staining of ANO5 was performed on 195 primary tumor samples obtained from patients with GC who underwent curative gastrectomy between 2011 and 2013 at our department.RESULTS Reverse transcription-quantitative polymerase chain reaction(PCR)and western blotting demonstrated high ANO5 mRNA and protein expression,respectively,in NUGC4 and MKN45 cells.In these cells,ANO5 silencing inhibited cell proliferation and induced apoptosis.In addition,the knockdown of ANO5 inhibited G1-S phase progression,invasion,and migration.The results of the microarray analysis revealed changes in the expression levels of several cyclin-associated genes,such as CDKN1A,CDK2/4/6,CCNE2,and E2F1,in ANO5-depleted NUGC4 cells.The expression of these genes was verified using reverse transcription-quantitative PCR.Immunohistochemical staining revealed that high ANO5 expression levels were associated with a poor prognosis.Multivariate analysis identified high ANO5 expression as an independent prognostic factor for 5-year survival in patients with GC(P=0

关键词： Anoctamin 5 Gastric cancer Cell cycle G1/S checkpoint Cell proliferation

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Laparoscopic cholecystectomy for gangrenous cholecystitis in around nineties: Two case reports

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World Journal of Clinical Cases 2021年第14期9卷 3424-3431页

作者： Hiroyuki Inoue Toshiya Ochiai Hidemasa Kubo Yusuke Yamamoto Ryo Morimura Hisashi Ikoma eigo otsuji Division of Digestive Surgery Department of SurgeryKyoto Prefectual University of MedicineKyoto 6028566Japan Department of Surgery North Medical Center Kyoto Prefectural University of MedicineKyoto 6292261Japan Division of Digestive Surgery Department of SurgeryKyoto Prefectural University of MedicineKyoto 6028566Japan

BACKGROUND Gangrenous cholecystitis is a form of acute cholecystitis which involves gangrenous alterations in the gallbladder wall and it often follows an acute and serious course.We herein report on two cases of very elderly people diagnosed early with gangrenous cholecystitis,who safely underwent laparoscopic cholecystectomy(LC)and both demonstrated a good outcome.CASE SUMMARY Case 1:An 89-year-old female.She underwent abdominal contrast-enhanced computed tomography(CECT)due to abdominal pain and diarrhea.Her gallbladder wall indicated the absence of contrast enhancement,thus leading to diagnosis of gangrenous cholecystitis and she therefore underwent LC.Although her gallbladder demonstrated diffuse necrosis and it was also partly perforated,she was able to be discharged without any serious complications.Case 2:A 91-year-old female.She made an emergency visit with a chief complaint of abdominal pain.Abdominal CECT revealed swelling of the gallbladder and an ambiguous continuity of the gallbladder wall.She was diagnosed with gangrenous cholecystitis and underwent LC.Her gallbladder had swelling and diffuse necrosis.Although her preoperative blood culture was positive,she showed a good outcome following surgery.CONCLUSION Although a definite diagnosis of gangrenous cholecystitis is difficult to make prior to surgery,if an early diagnosis can be made and appropriate treatment can be carried out,then even very elderly individuals may be discharged without major complications.

关键词： Gangrenous cholecystitis Laparoscopic cholecystectomy Elderly people Critical view of safety Tokyo guidelines 2018 Case report

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Laparoscopic and endoscopic co-operative surgery for nonampullary duodenal tumors

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World Journal of Gastroenterology 2016年第47期22卷 10424-10431页

作者： Daisuke Ichikawa Shuhei Komatsu Osamu Dohi Yuji Naito Toshiyuki Kosuga Kazuhiro Kamada Kazuma Okamoto Yoshito Itoh eigo otsuji Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Department of Gastroenterology and Hepatology Kyoto Prefectural University of Medicine

AIM To assess the safety and feasibility of laparoscopic and endoscopic co-operative surgery(LECS) for early nonampullary duodenal tumors.METHODS Twelve patients with a non-ampullary duodenal tumor underwent LECS at our hospital. One patient had two mucosal lesions in the duodenum. The indication for this procedure was the presence of duodenal tumors with a low risk for lymph node metastasis. In particular, the tumors included small(less than 10 mm) submucosal tumors(SMT) and epithelial mucosal tumors, such as mucosal cancers or large mucosal adenomas with malignant suspicion. The LECS procedures, such as full-thickness dissection for SMT and laparoscopic reinforcement after endoscopic submucosal dissection(ESD) for epithelial tumors, were performed for the 13 early duodenal lesions in 12 patients. Here we present the short-term outcomes and evaluate the safety and feasibility of this new technique.RESULTS Two SMT-like lesions and eleven superficial epithelial tumor-like lesions were observed. Seven and Six lesions were located in the second and third parts of the duodenum, respectively. All lesions were successfully resected en bloc. The defect in the duodenal wall was manually sutured after resection of the duodenal SMT. For epithelial duodenal tumors, the ulcer bed was laparoscopically reinforced via manual suturing after ESD. Intraoperative perforation occurred in two out of eleven epithelial tumor-like lesions during ESD; however, they were successfully laparoscopically repaired. The median operative time and intraoperative estimated blood loss were 322 min and 0 mL, respectively. Histological examination of the tumors revealed one adenoma with moderate atypia, ten adenocarcinomas, and two neuroendocrine tumors. No severe postoperative complications(Clavien-Dindo classification grade Ⅲ or higher) were reported in this series, but minor leakage secondary to pancreatic fistula occurred in one patient.CONCLUSION LECS can be a safe and minimally invasive treatment option for non-ampullary early duodenal tumors.

关键词： Non-ampullary 肿瘤 Laparoscopic 和内视镜的合作外科早十二指肠的癌症

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Granulocyte colony-stimulating factor-producing hepatocellular carcinoma with abrupt changes

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World Journal of Clinical Oncology 2016年第5期7卷 380-386页

作者： Hiroaki Nagata Shuhei Komatsu Wataru Takaki Tokunari Okayama Yasunori Sawabe Michiaki Ishii Mitsuo Kishimoto eigo otsuji Hiroshi Konosu Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Department of Surgery Ayabe City Hospital Department of Gastroenterology Ayabe City Hospital Department of Surgical Pathology Kyoto Prefectural University of Medicine

Granulocyte colony-stimulating factor(G-CSF)-producing tumor is one of the rare types of cancer clinically characterized by an elevated fever and white blood cell(WBC) increment. Although G-CSF producing tumors have been reported in several types of cancer including those of the lungs, cervix and bladder, G-CSF producing hepatocellular carcinoma is extremely rare. Here, we report the case of a rapidly growing and poorly differentiated hepatocellular carcinoma producing G-CSF. The patient showed symptoms of continuous high fever, stomach pain and cough, and high serum WBC counts, C-reactive protein(CRP) and G-CSF levels were found in laboratory tests. After a radical hepatectomy, the patient completely recovered from the above symptoms and inflammatory state. The serum levels of G-CSF were reduced to normal levels after radical surgery. An immunohistochemical analysis revealed the overexpression of G-CSF in the cytoplasm of certain hepatocellular carcinoma(HCC) cell. The patient's serum WBC, CRP and G-CSF levels remained within normal levels in the six months after surgery without recurrence. This is the 9^(th) case report of G-CSF producing hepatocellular carcinoma in English literature. We review the clinical characteristics of the G-CSF producing HCC and discuss a possible treatment strategy.

关键词： Granulocyte colony stimulating factor Granulocyte colony-stimulating factor producing tumor Hepatocellular carcinoma Immunohistochemistry Sarcomatous changes

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Intracellular chloride regulates the G_1/S cell cycle progression in gastric cancer cells

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World Journal of Gastrointestinal Oncology 2011年第8期3卷 119-122页

作者： Atsushi Shiozaki eigo otsuji Yoshinori Marunaka Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Department of Molecular Cell Physiology Graduate School of Medical Science Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Japan Institute for Food Education and Health St. Agnes' University Kyoto 602-8013 Japan

Recent studies show that ion channels/transporters play important roles in fundamental cellular functions. Several reports indicating the important roles of Cl- channels/transporters on cell proliferation suggest that the intracellular chloride concentration ([Cl-]i) regulated by them would be one of critical messengers. We investigated whether the [Cl-]i controls cell proliferation and cell cycle progression in human gastric cancer cells. Our studies indicated that furosemide, a blocker of Na+ /K+ /2Cl- cotransporter (NKCC), diminished cell growth by delaying the G1-S phase progression in gastric cancer cells with high expression and activity of NKCC. Furthermore, we found that the culture in the low Cl- medium (replacement of Cl- by NO3-) decreased the [Cl-]i and inhibited cell growth of gastric cancer cells and that this inhibition of cell growth was due to cell cycle arrest at the G0/G1 phase caused by diminutionof CDK2 and phosphorylated Rb. The culture of cells in the low Cl- medium significantly increased expressions of p21 mRNA and protein. In addition, the low Cl- medium induced phosphorylation of mitogen activated protein kinases (MAPKs). Treatment with an inhibitor of p38 or JNK significantly suppressed p21 upregulation caused by culture in a low Cl- medium and rescued gastric cancer cells from the low Cl- -induced G1 cell cycle arrest. These findings revealed that the [Cl-]i affects the cell proliferation via activation of MAPKs through upregulation of p21 in gastric cancer cells. Our results suggest that the [Cl-]i regulates important cellular functions in gastric cancer cells, leading to the development of novel therapeutic strategies.

关键词： Intracellular chloride Cell proliferation Cell cycle Gastric cancer Cl- channel Cl- transporter

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Cellular physiological approach for treatment of gastric cancer

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World Journal of Gastroenterology 2014年第33期20卷 11560-11566页

作者： Atsushi Shiozaki Daisuke Ichikawa eigo otsuji Yoshinori Marunaka Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Departments of Molecular Cell Physiology and Bio-Ionomics Graduate School of Medical Science Kyoto Prefectural University of Medicine Kyoto 602-8566 Japan Japan Institute for Food Education and Health St. Agnes’ University Kyoto 602-8013 Japan

Recent studies show that ion channels/transporters play important roles in fundamental cellular functions that would be involved in the cancer process. We review the evidence for their expression and functioning in human gastric cancer(GC), and evaluate the potential of cellular physiological approach in clinical management. Various types of ion channels, such as voltage-gated K+ channels, intracellular Cl- channels and transient receptor potential channels have been found to express in GC cells and tissues, and to control cell cycles. With regard to water channels, aquaporin 3 and 5 play an important role in the progression of GC. Regulators of intracellular pH, such as anion exchanger, sodiumhydrogen exchanger, vacuolar H+-ATPases and carbonic anhydrases are also involved in tumorigenesis of GC. Their pharmacological manipulation and gene silencing affect cellular behaviours, suggesting their potential as therapeutic targets for GC. Our studies indicate theintracellular Cl- concentration could act as a mediator of cellular signaling and control cell cycle progression in GC cells. Further, we demonstrate the cytocidal effects of hypotonic shock on GC cells, and indicate that the blockade of Cl- channels/transporters enhances these effects by inhibiting regulatory volume decrease. A deeper understanding of molecular mechanisms may lead to the discovery of these cellular physiological approaches as a novel therapeutic strategy for GC.

关键词： Gastric cancer Ion channels Water chan-nels Intrac

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Detection of fusion gene in cell-free DNA of a gastric synovial sarcoma

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World Journal of Gastroenterology 2018年第8期24卷 949-956页

作者： Shinpei Ogino Hirotaka Konishi Daisuke Ichikawa Junichi Hamada Katsutoshi Shoda Tomohiro Arita Shuhei Komatsu Atsushi Shiozaki Kazuma Okamoto Sanae Yamazaki Satoru Yasukawa Eiichi Konishi eigo otsuji Division of Digestive Surgery Department of Surgery Kyoto Prefectural University of Medicine Department of Surgical Pathology Kyoto Prefectural University of Medicine

Synovial sarcoma(SS) is genetically characterized by chromosomal translocation, which generates SYT-SSX fusion transcripts. Although SS can occur in any body part, primary gastric SS is substantially rare. Here we describe a detection of the fusion gene sequence of gastric SS in plasma cell-free DNA(cf DNA). A gastric submucosal tumor was detected in the stomach of a 27-year-old woman and diagnosed as SS. Candidate intronic primers were designed to detect the intronic fusion breakpoint and this fusion sequence was confirmed in intron 10 of SYT and intron 5 of SSX2 by genomic polymerase chain reaction(PCR) and direct sequencing. A locked nucleic acid(LNA) probe specificto the fusion sequence was designed for detecting the fusion sequence in plasma and the fusion sequence was detected in preoperative plasma cfD NA, while not detected in postoperative plasma cfD NA. This technique will be useful for monitoring translocation-derived diseases such as SS.

关键词： Fusion gene Gastric synovial sarcoma Plasma Cell free DNA

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Artificial intelligence in gastrointestinal cancer:Recent advances and future perspectives

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Artificial Intelligence in Gastroenterology 2020年第4期1卷 71-85页

作者： Michihiro Kudou Toshiyuki Kosuga eigo otsuji Division of Digestive Surgery Department of SurgeryKyoto Prefectural University of MedicineKyoto 602-8566Japan Department of Surgery Kyoto Okamoto Memorial HospitalKyoto 613-0034Japan

Artificial intelligence(AI)using machine or deep learning algorithms is attracting increasing attention because of its more accurate image recognition ability and prediction performance than human-aid analyses.The application of AI models to gastrointestinal(GI)clinical oncology has been investigated for the past decade.AI has the capacity to automatically detect and diagnose GI tumors with similar diagnostic accuracy to expert clinicians.AI may also predict malignant potential,such as tumor histology,metastasis,patient survival,resistance to cancer treatments and the molecular biology of tumors,through image analyses of radiological or pathological imaging data using complex deep learning models beyond human cognition.The introduction of AI-assisted diagnostic systems into clinical settings is expected in the near future.However,limitations associated with the evaluation of GI tumors by AI models have yet to be resolved.Recent studies on AI-assisted diagnostic models of gastric and colorectal cancers in the endoscopic,pathological,and radiological fields were herein reviewed.The limitations and future perspectives for the application of AI systems in clinical settings have also been discussed.With the establishment of a multidisciplinary team containing AI experts in each medical institution and prospective studies,AI-assisted medical systems will become a promising tool for GI cancer.

关键词： Artificial intelligence Gastric cancer Colorectal cancer Endoscopy Pathology Radiology

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Role of the Na^+/K^+/2Cl^- cotransporter NKCC1 in cell cycle progression in human esophageal squamous cell carcinoma

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World Journal of Gastroenterology 2014年第22期20卷 6844-6859页

作者： Atsushi Shiozaki Yoshito Nako Daisuke Ichikawa Hirotaka Konishi Shuhei Komatsu Takeshi Kubota Hitoshi Fujiwara Kazuma Okamoto Mitsuo Kishimoto Yoshinori Marunaka eigo otsuji Division of Digestive Surgery Department of SurgeryKyoto Prefectural University of MedicineKyoto 602-8566Japan Department of Pathology Kyoto PrefecturalUniversity of MedicineKyoto 602-8566Japan Departments of Molecular Cell Physiology and Bio-Ionomics Graduate School of Medical ScienceKyoto Prefectural University of MedicineKyoto 602-8566Japan Japan Institute for Food Education and Health St. Agnes’ UniversityKyoto 602-8013Japan

AIM:To investigate the role of Na+/K+/2Cl-cotransporter 1(NKCC1)in the regulation of genes involved in cell cycle progression and the clinicopathological significance of its expression in esophageal squamous cell carcinoma(ESCC).METHODS:An immunohistochemical analysis was performed on 68 primary tumor samples obtained from ESCC patients that underwent esophagectomy.NKCC1expression in human ESCC cell lines was analyzed by Western blotting.Knockdown experiments were conducted using NKCC1 small interfering RNA,and the effects on cell cycle progression were analyzed.The gene expression profiles of cells were analyzed by microarray analysis.RESULTS:Immunohistochemical staining showed that NKCC1 was primarily found in the cytoplasm of carcinoma cells and that its expression was related to the histological degree of differentiation of SCC.NKCC1 was highly expressed in KYSE170 cells.Depletion of NKCC1in these cells inhibited cell proliferation via G2/M phase arrest.Microarray analysis identified 2527 genes with altered expression levels in NKCC1depleted KYSE170.Pathway analysis showed that the top-ranked canonical pathway was the G2/M DNA damage checkpoint regulation pathway,which involves MAD2L1,DTL,BLM,CDC20,BRCA1,and E2F5.CONCLUSION:These results suggest that the expression of NKCC1 in ESCC may affect the G2/M checkpoint and may be related to the degree of histological differentiation of SCCs.We have provided a deeper understanding of the role of NKCC1 as a mediator and/or a biomarker in ESCC.

关键词： Na+/K+/2Cl-cotransporter 1 Esophageal cancer Cell

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Surgery for gastric cancer patients of age 85 and older: Multicenter survey

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World Journal of Gastroenterology 2017年第7期23卷 1215-1223页

作者： Hirotaka Konishi Daisuke Ichikawa Hiroshi Itoh Kenichiro Fukuda Naoki Kakihara Manabu Takemura Kaori Okugawa Kiyoshi Uchiyama Masashi Nakata Hiroshi Nishi Toshiyuki Kosuga Shuhei Komatsu Kazuma Okamoto eigo otsuji Division of Digestive Surgery Department of SurgeryKyoto Prefectural University of Medicine Department of Surgery North Medical CenterKyoto Prefectural University of Medicine Department of Surgery Saiseikai Shigaken Hospital Department of Surgery Japanese Red Cross Kyoto Daini Hospital Department of Surgery Osaka General Hospital of West Japan Railway Company Department of Surgery Omihachiman Community Medical Center Department of Surgery Kyoto Okamoto Memorial Hospital Department of Surgery Kyoto Yamashiro General Medical Center Department of Surgery Saiseikai Kyoto Hospital

AIM To investigate the surgical therapies for gastric cancer(GC) patients of age 85 or older in a multicenter survey.METHODS Therapeutic opportunities for elderly GC patients have expanded in conjunction with extended life expectancy. However, the number of cases encountered in a single institution is usually very small and surgical therapies for elderly GC patients have not yet been standardized completely. In the present study, a total of 134 GC patients of age 85 or older who underwent surgery in 9 related facilities were retrospectively investigated. The relationships between surgical therapies and clinicopathological or prognostic features were analyzed.RESULTS Eighty-nine of the patients(66%) presented with a comorbidity, and 26(19% overall) presented with more than two comorbidities. Radical lymphadenectomy was performed in 59 patients(44%), and no patient received pre- or post-operative chemotherapy. Forty of the patients(30%) experienced perioperative complications, but no surgical or perioperative mortality occurred. Laparoscopic surgery was performed in only 12 of the patients(9.0%). Univariate and multivariate analyses of the 113 patients who underwent R0 or R1 resection identified the factors of p T3/4 and limited lymphadenectomy as predictive of worse prognosis(HR = 4.68, P = 0.02 and HR =2.19, P = 0.05, respectively). Non-cancer-specific death was more common in c Stage Ⅰ patients than in c Stage Ⅱ or Ⅲ patients. Limited lymphadenectomy correlated with worse cancer-specific survival(P = 0.01), particularly in c Stage Ⅱ patients(P < 0.01). There were no relationships between limited lymphadenectomy and any comorbidities, except for cerebrovascular disease(P = 0.07). CONCLUSION N o n- c a n c e r- s p e c i f i c d e a t h w a s n o t n e g l i g i b l e, particularly in c Stage Ⅰ, and gastrectomy with radical lymphadenectomy appears to be an effective treatment for cS tage Ⅱ elderly GC patients.

关键词：胃的癌症老超过 85 外科有限 lymphadenectomy Multicenter 调查

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