MicroRNA-218靶向神经元表达发育下调基因9对胃癌细胞增殖和侵袭的影响  

Effect of microRNA-218 targeting neural precursor cell expressed developmentally down-regulated 9 on the proliferation and invasion of gastric cancer cells

作　　者：夏云展[1] 李荣振[1] 杨金花[2] 李琼[3] XIA Yunzhan;LI Rongzhen;YANG Jinhua;LI Qiong(Department of General Surgery,the People′s Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China;Department of Pathology,the People′s Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China;Department of Neurology,the People′s Hospital of Zhengzhou,Zhengzhou 450000,Henan Province,China)

机构地区：[1]郑州人民医院普通外科,河南郑州450000 [2]郑州人民医院病理科,河南郑州450000 [3]郑州人民医院神经内科,河南郑州450000

出　　处：《新乡医学院学报》2020年第3期219-224,共6页Journal of Xinxiang Medical University

基　　金：2018年度河南省科技攻关项目(编号:182102310355)。

摘　　要：目的观察microRNA-218(miR-218)对胃癌细胞SGC-7901增殖和侵袭的影响,并探讨其作用机制。方法体外培养人胃癌细胞SGC-7901,将对数生长期细胞分为空白对照组、阴性对照组和miR-218 mimics组,阴性对照组和miR-218 mimics组细胞分别转染阴性对照序列和miR-218 mimics,空白对照组细胞不进行转染。采用实时荧光定量聚合酶链反应检测3组细胞中miR-218表达,细胞计数试剂盒-8检测3组细胞培养24、48、72、96 h时的增殖活性;平板细胞克隆形成实验检测3组细胞克隆能力;Transwell实验检测3组细胞侵袭能力;Western blot法检测3组细胞中神经元表达发育下调基因9(NEDD9)蛋白、E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)、神经型钙黏附蛋白(N-cadherin)的表达。将SGC-7901细胞分为阴性对照组+NEDD93′非编码区(3′UTR)-Wt组(阴性对照序列、pCHECK2-NEDD93′UTR-Wt共转入SGC-7901细胞)、miR-218 mimics+NEDD93′UTR-Wt组(miR-218 mimics、pCHECK2-NEDD93′UTR-Wt共转入SGC-7901细胞)、阴性对照组+NEDD93′UTR-Mut组(阴性对照序列、pCHECK2-NEDD93′UTR-Mut共转入SGC-7901细胞)、miR-218 mimics+NEDD93′UTR-Mut组(miR-218 mimics、pCHECK2-NEDD93′UTR-Mut共转入SGC-7901细胞),常规培养48 h后收集各组细胞,采用发光检测仪测定各组细胞中荧光素酶活性,分析miR-218与NEDD9的靶向关系。结果与空白对照组和阴性对照组比较,miR-218 mimics组细胞中miR-218表达及培养24、48、72、96 h时细胞增殖抑制率显著升高(P<0.05),NEDD9蛋白表达及细胞克隆形成数量和侵袭细胞数量显著减少(P<0.05);空白对照组与阴性对照组细胞中以上各指标比较差异均无统计学意义(P>0.05)。miR-218 mimics+NEDD93′UTR-Wt组细胞中荧光素酶活性显著低于阴性对照组+NEDD93′UTR-Wt组、阴性对照组+NEDD93′UTR-Mut组和miR-218 mimics+NEDD93′UTR-Mut组(P<0.05)。与空白对照组和阴性对照组比较,miR-218 mimics组细胞中E-cadherin蛋白表达显著升高,Objective To investigate the effect of microRNA-218(miR-218)on the proliferation and invasion of gastric cancer cell line SGC-7901,and explore its mechanism.Methods Human gastric cancer cell line SGC-7901 cells were cultured in vitro.The logarithmic growth phase cells were divided into blank control group,negative control group and miR-218 mimics group.The cells in the negative control group and miR-218 mimics group were transfected with negative control sequence and miR-218 mimics respectively;the cells in the blank control group were not transfected.After transfection,the expression of miR-218 in the three group was detected by real-time fluorescence quantitative polymerase chain reaction;the proliferation activity of cells in the three group was detected by cell counting kit-8 assay at 24,48,72,96 h after cultured;the cloning ability of cells in the three group was detected by plate cell clone formation assay;the invasion ability of cells in the three group was detected by Transwell assay.The expressions of neural precursor cell expressed developmentally down-regulated 9(NEDD9),E-cadherin,vimentin and N-cadherin were detected by Western blot.The SGC-7901 cells were divided into negative control group+NEDD93′untranslated coding regions(3′UTR)-Wt group(the negative control sequence and pCHECK2-NEDD93′UTR-Wt were co transferred into SGC-7901 cells),miR-218 mimics+NEDD93′UTR-Wt group(miR-218 mimics and pCHECK2-NEDD93′UTR-Wt were co transferred into SGC-7901 cells),negative control group+NEDD93′UTR-Mut group(the negative control sequence and pCHECK2-NEDD93′UTR-Mut were co transferred into SGC-7901 cells)and miR-218 mimics+NEDD93′UTR-Mut group(the miR-218 mimics and pCHECK2-NEDD93′UTR-Mut were co-transferred into S GC-7901 cells),after 48 hours of conventional culture,the cells in each group were collected,and the luciferase activity of the cells in each group was measured by light-emitting detector to analyze the targeting relationship between miR-218 and NEDD9.Results Compared with the blank cont

关 键 词：胃癌 增殖 侵袭 microRNA-218 神经元表达发育下调基因9 

分 类 号：R735.2[医药卫生—肿瘤]