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目的:探究微小核糖核酸(micro RNA,mi RNA)-143在胃癌细胞增殖与迁移中的作用与机制。方法:三对肿瘤组织及配对正常组织选自2015年1月至2015年5月天津医科大学肿瘤医院行胃癌根治术的患者,所有患者术前均未予任何放化疗,肿瘤组织病理检测证实均为中分化胃腺癌,癌旁正常组织未见癌细胞浸润。采用Western blot检测胃癌、癌旁正常组织和SGC7901胃癌细胞中鸟成红细胞增多症癌基因-3(avian erythroblastosis oncogene B-3,ERBB3)的蛋白表达水平,逆转录定量聚合酶链式反应(reverse transcription quantitative polymerase chain reaction,RT-q PCR)检测ERBB3 m RNA和mi R-143的表达水平,生物信息学软件预测mi R-143的靶基因,荧光素酶报告基因实验验证靶基因,Transwell迁移实验和Ed U增殖实验分别检测用mi R-143 mimics/inhibitor/NC mimics/inhibitor转染SGC7901细胞后对其迁移和增殖能力的影响。结果:与癌旁正常组织相比,胃癌组织ERBB3蛋白表达水平明显上升,m RNA水平升高差异远不及蛋白显著,mi R-143表达水平显著下降;生物信息学软件预测ERBB3 m RNA的3’非翻译区(untranslated regions,UTR)有1个mi R-143的结合位点,荧光素酶报告基因实验证实靶点存在;体外实验通过细胞转染上调mi R-143后ERBB3蛋白表达水平明显下降,而下调mi R-143后明显升高;Transwell迁移实验和Ed U增殖实验分别显示mi R-143过表达后细胞迁移和增殖能力显著减弱,而下调mi R-143后显著增强。结论:mi R-143可通过抑制ERBB3的表达抑制胃癌细胞的增殖与迁移。
Objective: To explore the role and mechanism of microRNA (miRNA) -143 in the proliferation and migration of gastric cancer cells. Methods: Three pairs of tumor tissues and paired normal tissues were selected from patients who underwent radical gastrectomy at Cancer Hospital of Tianjin Medical University from January 2015 to May 2015. All the patients were treated with radiotherapy and chemotherapy before operation, and confirmed by histopathological examination For moderately differentiated gastric adenocarcinoma, adjacent normal tissue no cancer cell infiltration. Western blot was used to detect the protein expression of avian erythroblastosis oncogene B-3 (ERBB3) in gastric cancer tissues, adjacent non-cancerous tissues and SGC7901 gastric cancer cells, and the reverse transcription quantitative polymerase chain reaction quantitative RT-qPCR) was used to detect the expression of ERBB3 mRNA and mi R-143. Bioinformatics software was used to predict the target gene of mi R-143, luciferase reporter assay, target gene Transwell migration assay and Ed U proliferation assay SGC7901 cells transfected with mi R-143 mimics / inhibitor / NC mimics / inhibitor were detected after migration and proliferation. Results: The expression of ERBB3 protein in gastric cancer tissues was significantly higher than that in adjacent normal tissues. The difference of m RNA level was far less than that of protein and the expression of mi R-143 was significantly decreased. Bioinformatics software predicted the 3 ’ There was one mi R-143 binding site in untranslated region (UTR). Luciferase reporter assay confirmed the target site. ERBB3 protein expression was significantly down-regulated after mi R-143 transfection in vitro , While downregulation of mi R-143 was significantly increased. Transwell migration assay and Ed U proliferation assay showed that mi R-143 overexpression significantly decreased cell migration and proliferation, but down-regulated mi R-143 significantly. Conclusion: mi R-143 can inhibit the proliferation and migration of gastric cancer cells by inhibiting the expression of ERBB3.