miR-29a靶向调控IFITM3表达对肝癌HepG2细胞生长、增殖和凋亡的影响  

Effect of miR-29a targeted-regulating IFITM3 expression on growth,proliferation and apoptosis of hepatocellular carcinoma HepG2 cells

作　　者：潘春鹏 李峰[1] 杨孙虎[1] PAN Chun-peng;LI Feng;YANG Sun-hu(Department of General Surgery,Shanghai Ninth People's Hospital,Shanghai JiaoTong University School of Medicine,Shanghai 200433,China)

机构地区：[1]上海交通大学医学院附属第九人民医院普外科,上海200433

出　　处：《临床和实验医学杂志》2019年第4期369-373,共5页Journal of Clinical and Experimental Medicine

摘　　要：目的探究微小RNA-29a(miR-29a)靶向调控干扰素诱导跨膜蛋白3(IFITM3)表达对肝癌HepG2细胞生长、增殖和凋亡的影响。方法采用实时荧光定量PCR(qRT-PCR)检测肝癌细胞HepG2和正常肝细胞株LO2中miR-29a的表达水平;采用脂质体法将miR-29a minmic及阴性对照转染至HepG2细胞,并分为过表达组和对照组,转染48 h后用QRCR法检测两组miR-29a水平;采用MTT法和流式细胞术检测各组细胞增殖及凋亡情况; Western blotting检测各组Bax、caspase-3凋亡相关基因及IFITM3的表达;采用双荧光素酶报告基因实验验证miR-29a对IFITM3的靶向调控作用。结果肝癌HepG2细胞miR-29a水平低于正常LO2细胞(P <0. 05);过表达组HepG2细胞miR-29a表达低于对照组(P <0. 05);过表达组HepG2细胞增殖率低于对照组(P <0. 05),凋亡率及Bax、caspase-3、IFITM3表达高于对照组(P <0. 05); miR-29a可显著抑制野生型FITM3-3'UTR质粒转染细胞的荧光素酶活性(P <0. 05),但其对突变型FITM3-3'UTR质粒转染细胞的荧光素酶活性无显著影响(P> 0. 05)。结论 miR-29a在肝癌中表达降低,可靶向调控IFITM3表达,抑制肝癌细胞生长、增殖,并促进其凋亡,可作为肝癌的潜在治疗靶点。Objective To investigate the effect of miR-29a targeted-regulating interferon induced transmembrane protein 3(IFITM3)expression on growth,proliferation and apoptosis of hepatocellular carcinoma HepG2 cells.Methods qRT-PCR was used to detect the expression of miR-29a in HepG2 cells and normal liver cell line LO2.miR-29a mimic and blank liposome(control purpose)were transfected into HepG2 cells by lipofectamine method,namely overexpression group and control group.The expression of miR-29a was detected by QRCR method after 48 h of transfection.Proliferation and apoptosis of cells in each group were detected by MTT assay and flow cytometry.Bax and caspase apoptosis-related genes and IFITM3 expression were detected by Western blotting;dual-luciferase reporter gene assay was used to verify targeted regulation of miR-29a on IFITM3.Results The level of miR-29a in HepG2 cells was lower than in normal LO2 cells(P﹤0.05).The expression of miR-29a in HepG2 cells in overexpression group was lower than in control group(P﹤0.05).The proliferation rate of HepG2 cells in overexpression group was lower than in control group(P﹤0.05)while apoptosis rate,expression of Bax,caspase-3 and IFITM3 were higher than in control group(P﹤0.05).miR-29a can significantly inhibit luciferase activity of wild-type FITM3-3'UTR plasmid transfected cells(P﹤0.05),but had no significant effect on that of mutant FITM3-3'UTR plasmid transfected cells(P﹥0.05).Conclusion Hepatocarcinoma cells tend to have lower miR-29a,whose expression can positively regulate expression of IFITM3,which can inhibit growth and proliferation of hepatocellular carcinoma cells and promote apoptosis,indicating a potential therapeutic target in hepatocarcinoma.

关 键 词：肝癌HEPG2细胞 miR-29a 干扰素诱导跨膜蛋白3 靶向调控 

分 类 号：R735.7[医药卫生—肿瘤]