miR-34c-5p过表达抑制胃癌细胞侵袭和迁移  

Overexpression of miR-34c-5p inhibits invasion and migration of gastric cancer cells

作　　者：赵媛 李冰 陶月佳 黄允宁[2] 徐远义[1] ZHAO Yuan;LI Bing;TAO Yue-jia;HUANG Yun-ning;XU Yuan-yi(Dept of Pathology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China;Dept of Gastrointestinal Surgery,People’s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院病理系,宁夏银川750004 [2]宁夏回族自治区人民医院胃肠外科,宁夏银川750004

出　　处：《中国药理学通报》2022年第12期1830-1835,共6页Chinese Pharmacological Bulletin

基　　金：宁夏回族自治区重点研发项目(No 2019BEG03007);宁夏自然科学基金资助项目(No 2020AAC03184,2022AAC03368)。

摘　　要：目的研究微小RNA(miR)-34c-5p在胃癌组织中的表达及调控胃癌细胞侵袭和迁移机制。方法利用实时荧光定量聚合酶链式反应(RT-qPCR)检测人胃癌组织及胃癌细胞中miR-34c-5p mRNA表达;在人胃癌细胞中构建miR-34c-5p过表达模型,倒置荧光显微镜观察慢病毒转染情况,RT-qPCR验证转染效率。利用划痕实验、Transwell实验检测过表达miR-34c-5p对胃癌细胞侵袭和迁移的影响;通过Western blot检测各组胃癌细胞中MAP2K1、E-钙黏蛋白(E-cadherin)及波形蛋白(Vimentin)的表达情况。双萤光素酶报告基因检测MAP2K1是否是miR-34c-5p的靶基因。结果miR-34c-5p mRNA在10例人胃癌组织中的表达明显低于癌旁组织(P<0.05);miR-34c-5p mRNA在胃癌HGC-27细胞中的表达明显低于正常胃黏膜GES-1细胞(P<0.01);与空白对照组、miR-34c-5p-NC组比较,miR-34c-5p-mimics组miR-34c-5p mRNA表达水平明显升高(P<0.05);划痕实验及Transwell实验结果表明,过表达miR-34c-5p明显抑制了胃癌细胞的迁移和侵袭能力(P<0.05,P<0.01);Western blot检测结果表明,过表达miR-34c-5p后胃癌HGC-27细胞中MAP2K1、Vimentin蛋白表达水平明显降低,E-cadherin蛋白表达水平明显升高(P<0.05);此外,与mimic-NC+miR-34c-5p-mimics组相比,转染pmiR-MAP2K1-WT质粒细胞的萤光素酶活性降低(P<0.01),而转染pmiR-MAP2K1-Mut质粒的萤光素酶活性无改变。结论miR-34c-5p在胃癌组织和胃癌细胞中低表达,可能通过靶向调控MAP2K1表达抑制胃癌细胞侵袭和迁移。Aim To study the expression of microRNA(miR-34c-5p)in gastric cancer tissues and the mechanism of regulating the invasion and migration of gastric cancer cells.Methods The expression of miR-34c-5p mRNA in human gastric cancer tissues and cells was detected by quantitative real-time polymerase chain reaction(RT-qPCR).An overexpression model of miR-34c-5p was constructed in human gastric cancer cells.The transfection of lentivirus was observed under inverted fluorescence microscope,and the transfection efficiency was verified by RT-qPCR.Scratch test and Transwell assay were used to detect the effects of overexpression of miR-34c-5p on invasion and migration of gastric cancer cells.Western blot was used to detect the expression of MAP2K1,E-cadherin and Vimentin in gastric cancer cells of each group.Dual-luciferase reporter assay was employed to detect whether MAP2K1 was a target gene of miR-34c-5p.Results The expression of miR-34c-5p mRNA in 10 human gastric cancer tissues was significantly lower than that in adjacent tissues(P<0.05).The expression of miR-34c-5p mRNA in gastric cancer HGC-27 cell was significantly lower than that in normal gastric mucosa GES-1 cell(P<0.01).Compared with blank control group and miR-34c-5p-NC group,miR-34c-5p-mimics group significantly increased its miR-34c-5p mRNA expression level(P<0.05).The results of scratch test and Transwell experiment showed that overexpression of miR-34c-5p significantly inhibited the migration and invasion ability of gastric cancer cells(P<0.05,P<0.01).Western blot results showed that the protein expression levels of MAP2K1 and Vimentin significantly decreased and E-cadherin markedly increased in gastric cancer HGC-27 cells after overexpression of miR-34c-5p(P<0.05).In addition,compared with mimic-NC+miR-34c-5p-mimics group,the lucifase activity of pmiR-MAP2K1-WT plasmid decreased(P<0.01),while the lucifase activity of pmiR-MAP2K1-MUT plasmid was not changed.Conclusions The low expression of miR-34c-5p in gastric cancer tissues and cells may inhibit the invas

关 键 词：胃癌 微小RNA-34c-5p MAP2K1 细胞侵袭 细胞迁移 E-CADHERIN VIMENTIN 

分 类 号：R342.2[医药卫生—基础医学] R394.2R735.2R735.202.2