MiR-1184靶向胰岛素样生长因子结合蛋白4调控肝癌细胞增殖、迁移及凋亡

MiR-1184 targets insulin-like growth factor binding protein 4 to regulate the proliferation,migration and apoptosis of hepatocellular carcinoma cells

目的:探究miR-1184对人肝癌细胞增殖、迁移及凋亡功能的影响及其机制。方法:实时荧光定量PCR(qRT-PCR)检测人肝癌细胞系(Huh7、Hep3B、HepG2、HCCLM3、SK-HEP1和MHCC97H)和正常人肝细胞(LO2)内miR-1184的表达,分别用miR-1184 mimic和miR-1184 NC转染Huh7和Hep3B细胞后用qRT-PCR法检测miR-1184的表达并完成CCK-8、Transwell实验及流式细胞术实验。利用Targetscan Human 8.0预测了miR-1184的靶基因,并通过qRT-PCR和蛋白印迹法(Western Blot)检测转染后细胞内IGFBP4表达水平变化。结果:miR-1184在肝癌细胞系中低表达(P<0.05);过表达miR-1184后,肝癌细胞增殖(P<0.01)、迁移减弱(P<0.01),但凋亡能力增强(P<0.05);过表达miR-1184后,IGFBP4 mRNA及蛋白表达水平增强(P<0.05)。结论:miR-1184可通过调控IGFBP4的表达水平抑制肝癌细胞增殖、迁移并促进凋亡。

Objective:To explore the effect and mechanism of miR-1184 on proliferation,migration and apoptosis of human hepatocellular carcinoma(HCC)cells.Methods:Real-time quantitative PCR(qRT-PCR)was adopted to detect the expression of miR-1184 in hepatocellular carcinoma cell lines(Huh7,Hep3B,HepG2,HCCLM3,SK-HEP1,and MHCC97H)and normal liver cells(LO2).After transfection of Huh7 and Hep3B cells with miR-1184 mimic and miR-1184 NC,the expression of miR-1184 was also detected by qRT-PCR,and CCK-8,Transwell assay and apoptosis assay were performed as well.The target gene of miR-1184 was predicted using Targetscan Human 8.0.The qRT-PCR and Western Blot were used to detect the expression of IGFBP4 in transfected cells.Results:The expression of miR-1184 was low in liver cancer cell lines,and the difference was statistically significant(P<0.05).After overexpression of miR-1184,proliferation(P<0.01)and migration of HCC cells were decreased(P<0.01),but apoptosis was enhanced(P<0.05).After overexpression of miR-1184,the mRNA and protein expression levels of IGFBP4 were increased(P<0.01).miR-1184 can inhibit the proliferation and migration but promote apoptosis of HCC cells by regulating the expression level of IGFBP4.