微小RNA-218-5p对膀胱癌细胞增殖和凋亡的影响及分子机制  

Effect of microRNA-218-5p on the proliferation and apoptosis of bladder cancer cells and its molecular mechanism

作　　者：孙鑫波 陈从波 陈绪华 黄力 SUN Xinbo;CHEN Congbo;CHEN Xuhua;HUANG Li(Department of Urology,Hubei Medical College Affiliated to Shiyan Taihe Hospital,Shiyan 442000,Hubei,China;Department of Dermatology,Hubei Medical College Affiliated to Shiyan Taihe Hospital,Shiyan 442000,Hubei,China)

机构地区：[1]十堰市太和医院(湖北医药学院附属医院)泌尿外科,湖北十堰4420000 [2]十堰市太和医院(湖北医药学院附属医院)皮肤科,湖北十堰4420000

出　　处：《癌症进展》2021年第5期458-462,518,共6页Oncology Progress

摘　　要：目的探讨miRNA-218-5p对膀胱癌细胞增殖和凋亡的影响及其作用机制。方法将BIU-78膀胱癌细胞分为miRNA-con组(转染miRNA-con)、miRNA-218-5p组(转染miRNA-218-5p mimics)、anti-miRNA-con组(转染anti-miRNA-con)、anti-miRNA-218-5p组(转染anti-miRNA-218-5p)、si-con组(转染si-con)、si-98序列相似的家族成员A(FAM98A)组(转染si-FAM98A)、miRNA-218-5p+pcDNA-con组(共转染miRNA-218-5p和pcDNAcon)、miRNA-218-5p+pcDNA-FAM98A组(共转染miRNA-218-5p和pcDNA-FAM98A)。采用定量逆转录聚合酶链反应(qRT-PCR)检测miRNA-218-5p和FAM98A mRNA的表达水平;蛋白质印迹法(Western blot)检测蛋白表达;MTT法检测细胞增殖率;流式细胞术检测细胞凋亡率;双荧光素酶报告基因检测实验检测荧光活性。结果相较于正常膀胱组织细胞SV-HUC-1,膀胱癌细胞BIU-78、T24、EJ中FAM98A mRNA和蛋白表达水平均升高,miRNA-218-5p表达水平均降低(P﹤0.05)。过表达miRNA-218-5p和敲减FAM98A均可抑制膀胱癌细胞BIU-78增殖,诱导细胞凋亡,抑制cyclin D1、B细胞淋巴瘤/白血病-2(Bcl-2)表达,促进Bcl-2相关X蛋白(BAX)表达。miRNA-218-5p靶向负调控FAM98A的表达,过表达FAM98A能逆转miRNA-218-5p对膀胱癌细胞BIU-78的增殖抑制和凋亡促进作用。过表达miRNA-218-5p抑制磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(AKT)信号通路中p-PI3Kp85和p-AKT蛋白的表达,过表达FAM98A能逆转miRNA-218-5p对p-PI3Kp85和p-AKT表达的抑制作用。结论miRNA-218-5p可抑制膀胱癌细胞增殖,诱导细胞凋亡,其机制可能与FAM98A及PI3K/AKT信号通路有关,将可为膀胱癌的预防和治疗提供新靶点。Objective To investigate the effect of miRNA-218-5p on proliferation and apoptosis of bladder cancer cells and its underlying mechanism.Method The BIU-78 bladder cancer cells were divided into miRNA-con group(transfected with miRNA-con),miRNA-218-5p group(transfected with miRNA-218-5p mimics),anti-miRNA-con group(transfected with anti-miRNA-con),anti-miRNA-218-5p group(transfected with anti-miRNA-218-5p),si-con group(transfected with si-con),si-family with sequence similarity 98,member A(FAM98 A) group(transfected with siFAM98 A),miRNA-218-5p+pcDNA-con group(co-transfected with miRNA-218-5p and pcDNA-con),miRNA-218-5p+pcDNA-FAM98 A group(co-transfected with miRNA-218-5p and pcDNA-FAM98 A).The expressions of miRNA-218-5p and FAM98 A mRNA were detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR);protein expression was detected by Western blot;cell viability was detected by MTT assay;cell apoptosis was detected by flow cytometry;fluorescence activity was detected by dual-luciferase reporter assay.Result Compared with normal bladder tissue cells SV-HUC-1,the expression levels of FAM98 A mRNA and protein in bladder cancer cells BIU-78,T24,and EJ were sigmficantly increased while the expression of miRNA-218-5p was significantly decreased(P<0.05).Overexpression of miRNA-218-5p and knockdown of FAM98 A inhbited the proliferation of bladder cancer cell line BIU-78 and induced their apoptosis,inhibited cyclin D1,B-cell lymphoma/leukemia-2(Bcl-2) expression,and promoted Bcl-2 related protein X(BAX) expression.miRNA-218-5p could target FAM98 A and negatively regulates its expression.Overexpression of FAM98 A could reverse the proliferation inhibition and apoptosis-promoting effect of miRNA-218-5p on bladder cancer cell line BIU-78.Overexpression of miRNA-218-5p could inhibit p-PI3 Kp85 and p-AKT in the phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(AKT) signaling pathway,and overexpression of FAM98 A could reverse this effect.Conclusion miRNA-218-5p could inhibit the proliferation of bladd

关 键 词：miRNA-218-5p 98序列相似的家族成员A 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 膀胱癌 增殖 凋亡 

分 类 号：R737.14[医药卫生—肿瘤]