Tae-Su Han1, Dominic Chih-Cheng Voon2, Hiroko Oshima3, Mizuho Nakayama3, Kanae Echizen4, Eri Sakai5, Zachary Wei Ern Yong5, Kazuhiro Murakami5, Liang Yu6, Toshinari Minamoto7, Chan-Young Ock8, Brendan J Jenkins6, Seong-Jin Kim8, Han-Kwang Yang9, Masanobu Oshima10. 1. Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan; AMED-CREST, AMED, Japan Agency for Medical Research and Development, Tokyo, Japan; Biotherapeutics Translational Research Center, Division of Biomedical Science, Korea Research Institute of Bioscience and Biotechnology, Daejeon, Korea. 2. Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan; Innovative Cancer Model Research Unit, Institute for Frontier Science Initiative, Kanazawa University, Kanazawa, Japan. Electronic address: dvoon@staff.kanazawa-u.ac.jp. 3. Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan; WPI Nano-Life Science Institute (Nano-LSI), Kanazawa University, Kanazawa, Japan. 4. Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan; AMED-CREST, AMED, Japan Agency for Medical Research and Development, Tokyo, Japan. 5. Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan. 6. Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Monash University, Clayton, Australia; Department of Molecular Translational Science, School of Clinical Sciences, Monash University, Clayton, Australia. 7. Division of Translational and Clinical Oncology, Cancer Research Institute, Kanazawa University, Kanazawa, Japan. 8. Theragen Etex Bio Institute, Suwon, Korea; Precision Medicine Research Center, Advanced Institutes of Convergence Technology and Department of Transdisciplinary Studies, Seoul National University, Suwon, Korea. 9. Department of Surgery and Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea. 10. Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan; AMED-CREST, AMED, Japan Agency for Medical Research and Development, Tokyo, Japan; WPI Nano-Life Science Institute (Nano-LSI), Kanazawa University, Kanazawa, Japan. Electronic address: oshimam@staff.kanazawa-u.ac.jp.
Abstract
BACKGROUND & AIMS: Gastritis is associated with development of stomach cancer, but little is known about changes in microRNA expression patterns during gastric inflammation. Specific changes in gene expression in epithelial cells are difficult to monitor because of the heterogeneity of the tissue. We investigated epithelial cell-specific changes in microRNA expression during gastric inflammation and gastritis-associated carcinogenesis in mice. METHODS: We used laser microdissection to enrich epithelial cells from K19-C2mE transgenic mice, which spontaneously develop gastritis-associated hyperplasia, and Gan mice, which express activated prostaglandin E2 and Wnt in the gastric mucosa and develop gastric tumors. We measured expression of epithelial cell-enriched microRNAs and used bioinformatics analyses to integrate data from different systems to identify inflammation-associated microRNAs. We validated our findings in gastric tissues from mice and evaluated protein functions in gastric cell lines (SNU-719, SNU-601, SNU-638, AGS, and GIF-14) and knockout mice. Organoids were cultured from gastric corpus tissues of wild-type and miR-135b-knockout C57BL/6 mice. We measured levels of microRNAs in pairs of gastric tumors and nontumor mucosa from 28 patients in Japan. RESULTS: We found microRNA 135b (miR-135B) to be the most overexpressed microRNA in gastric tissues from K19-C2mE and Gan mice: levels increased during the early stages of gastritis-associated carcinogenesis. Levels of miR-135B were also increased in gastric tumor tissues from gp130F/F mice and patients compared with nontumor tissues. In gastric organoids and immortalized cell lines, expression of miR-135B was induced by interleukin 1 signaling. K19-C2mE mice with disruption of Mir-135b developed hyperplastic lesions that were 50% smaller than mice without Mir-135b disruption and had significant reductions in cell proliferation. Expression of miR-135B in gastric cancer cell lines increased their colony formation, migration, and sphere formation. We identified FOXN3 and RECK messenger RNAs (mRNAs) as targets of miR-135B; their knockdown reduced migration of gastric cancer cell lines. Levels of FOXN3 and RECK mRNAs correlated inversely with levels of miR-135B in human gastric tumors and in inflamed mucosa from K19-C2mE mice. CONCLUSIONS: We found expression of miR-135B to be up-regulated by interleukin L1 signaling in gastric cancer cells and organoids. miR-135B promotes invasiveness and stem-cell features of gastric cancer cells in culture by reducing FOXN3 and RECK messenger RNAs. Levels of these messenger RNA targets, which encode tumor suppressor, are reduced in human gastric tumors.
BACKGROUND & AIMS:Gastritis is associated with development of stomach cancer, but little is known about changes in microRNA expression patterns during gastric inflammation. Specific changes in gene expression in epithelial cells are difficult to monitor because of the heterogeneity of the tissue. We investigated epithelial cell-specific changes in microRNA expression during gastric inflammation and gastritis-associated carcinogenesis in mice. METHODS: We used laser microdissection to enrich epithelial cells from K19-C2mE transgenic mice, which spontaneously develop gastritis-associated hyperplasia, and Gan mice, which express activated prostaglandin E2 and Wnt in the gastric mucosa and develop gastric tumors. We measured expression of epithelial cell-enriched microRNAs and used bioinformatics analyses to integrate data from different systems to identify inflammation-associated microRNAs. We validated our findings in gastric tissues from mice and evaluated protein functions in gastric cell lines (SNU-719, SNU-601, SNU-638, AGS, and GIF-14) and knockout mice. Organoids were cultured from gastric corpus tissues of wild-type and miR-135b-knockout C57BL/6 mice. We measured levels of microRNAs in pairs of gastric tumors and nontumor mucosa from 28 patients in Japan. RESULTS: We found microRNA 135b (miR-135B) to be the most overexpressed microRNA in gastric tissues from K19-C2mE and Gan mice: levels increased during the early stages of gastritis-associated carcinogenesis. Levels of miR-135B were also increased in gastric tumor tissues from gp130F/F mice and patients compared with nontumor tissues. In gastric organoids and immortalized cell lines, expression of miR-135B was induced by interleukin 1 signaling. K19-C2mE mice with disruption of Mir-135b developed hyperplastic lesions that were 50% smaller than mice without Mir-135b disruption and had significant reductions in cell proliferation. Expression of miR-135B in gastric cancer cell lines increased their colony formation, migration, and sphere formation. We identified FOXN3 and RECK messenger RNAs (mRNAs) as targets of miR-135B; their knockdown reduced migration of gastric cancer cell lines. Levels of FOXN3 and RECK mRNAs correlated inversely with levels of miR-135B in humangastric tumors and in inflamed mucosa from K19-C2mE mice. CONCLUSIONS: We found expression of miR-135B to be up-regulated by interleukin L1 signaling in gastric cancer cells and organoids. miR-135B promotes invasiveness and stem-cell features of gastric cancer cells in culture by reducing FOXN3 and RECK messenger RNAs. Levels of these messenger RNA targets, which encode tumor suppressor, are reduced in humangastric tumors.
Authors: Abdul Q Khan; Eiman I Ahmed; Noor R Elareer; Kulsoom Junejo; Martin Steinhoff; Shahab Uddin Journal: Cells Date: 2019-08-05 Impact factor: 6.600