The effect of microRNA-21-siRNA-lentivirus on biological behaviors in human hepatic cancer cell line HepG2

Abstract

Objective To study the effect of lentivirus-mediated microRNA-21 RNAi on biological behaviors in human hepatic cancer cells.Methods MicroRNA-21 specific siRNA gene was synthesized and cloned into the recombinant lentiviral vector,pGCSIL-GFP.HepG2 cells were infected by microRNA-21 siRNA recombinant lentivirus (miR-21-siRNA-Ⅳ).The HepG2 cells were devided into SI group,NC group and N group in vitro.The expression of the targets of miR-21 was detected by RT-PCR.Cell growth was analyzed by MTT assay.The invasion was dectected by tmnswell method.Apoptosis was detected by Hoechst33258.BALB/c nude mice were randomly divided into SI group and NC group.The growth of transplant tumors in BALB/c nude mice were observed.Results ( 1 ) The expression level of miR-21 was inhibited significantly by miR-21-siRNA-lⅣ.(2) The proliferation of HepG2 was also markedly suppressed in MTT at the 96 h point ( P =0.0031,P ＜ 0.05 ).(3) The number of cells that migrated through the chamber of SI group decreased ( P =0.0004,P ＜ 0.05 ).(4) The cell apopotosis in SI group increased markedly.In addition,the caspase 3 mRNA significantly increased ( P =0.0002,P ＜ 0.05 ).( 5 ) Tumor growth curve was not statistically different between groups ( P =0.0002,P ＜ 0.05 ).Conclusions MicroRNA-21 specific siRNA suppresses the proliferation and migration of HepG2 cells and induces tumor cell apoptosis inhibiting the growth of transplanted tumor in Balb/c nude mice. Key words: Carcinoma, hepatocellular; RNA, small interfering; Animal experimentation; MicroRNA-21