微小RNA188-3p靶向调节N-myc下游调节基因1抑制肝癌细胞的增殖  

MicroRNA-188-3p and its target gene N-myc downstream regulated gene 1 on proliferation activity of hepatocarcinoma cells

作　　者：孔晓宇[1] 罗振钊[2] 刘显畅 魏礼清[2] Kong Xiaoyu;Luo Zhenzhao;Liu Xianchang;Wei Liqing(Department of Hepatobilary&Pancreatic Surgery,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China;Department of Laboratory,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430014,China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院肝胆外科,430014 [2]华中科技大学同济医学院附属武汉中心医院检验科,430014

出　　处：《中华实验外科杂志》2020年第11期2094-2096,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省卫生健康科研基金(WJ2019M031)。

摘　　要：目的探讨微小RNA(miRNA,miR)-188-3p在肝癌发生发展中的作用及其机制。方法收集2018年2月至2019年10月武汉市中心医院肝胆胰外科经手术切除并病理证实的21例原发性肝癌组织标本及对应的13例癌旁组织标本,细胞培养肝癌细胞,用生物信息学预测miR-188-3p和N-myc下游调节基因1(NDRG1)的靶向匹配关系,并用双荧光素酶报告基因系统鉴定。Lipo3000转染miR-188-3p摸拟物后,实时荧光定量聚合酶链反应(Real-time qPCR)检测miR-188-3p和NDRG1 mRNA表达;蛋白质印迹法(Western blot)检测NDRG1蛋白表达;细胞计数试剂盒(CCK-8)实验检测细胞增殖。组间比较采用t检验。结果miR-188-3p和NDRG1匹配良好,miR-188-3p能够结合NDRG1 mRNA 3’端非编码区(3’UTR),并有效抑制其mRNA及蛋白表达水平(0.56±0.04比1.12±0.13,t=10.777,P<0.01;0.43±0.07比0.88±0.15,t=9.123,P<0.05)。miR-188-3p能够负向调控NDRG1表达和肝癌细胞增殖(0.76±0.07比1.26±0.19,t=7.509,P<0.05)。结论miR-188-3p可以通过下调靶向癌基因NDRG1的表达来抑制肝癌细胞的增殖。Objective To investigate the effect of microRNA(miRNA,miR)-188-3p on N-myc downstream regulated gene 1(NDRG1)and its role in the proliferation activity of hepatocarcinoma cells(HCC).Methods Collected 21 cases of HCC tissue specimens and 13 cases of adjacent cancer tissues that were surgically resected and pathologically confirmed by the Department of Hepatobiliary and Pancreatic Surgery,Wuhan Central Hospital from February 2018 to October 2019,HepG2 cells were cultured in vitro.In this study,the miR-188-3p was predicted with bioinformatics and identified with dual luciferase report system.Expression of miR-188-3p and NDRG1 was determined with real-time quantitative polymerase chain reaction(Real-time PCR)and Western blotting after transfection of miR-188-3p.The proliferation of HepG2 cells was detected in vitro by CCK-8 assay.Results MiR-188-3p can inhibit NDRG1 expression by binding to 3’untranslated regions(3’UTR)of NDRG1 mRNA.miR-188-3p can negatively control the mRNA and protein expression of NDRG1(0.56±0.04 vs.1.12±0.13,t=10.777,P<0.01、0.43±0.07 vs.0.88±0.15,t=9.123,P<0.05).The proliferation of HepG2 cells was negatively controlled by miR-188-3p(0.76±0.07 vs.1.26±0.19,t=7.509,P<0.05).Conclusion MiR-188-3p could effectively inhibit the proliferation of HCC through targting NDRG1,suggesting miR-188-3p could serve as an innovative therapeutic strategy in terms of HCC.

关 键 词：肝癌 增殖 微小RNA-188-3p N-myc下游调节基因1 

分 类 号：R735.7[医药卫生—肿瘤]