LV-hsa-mir-34a enhance the inhibitory effects of Doxorubicin on hepatocellular carcinoma cells / 中华普通外科杂志

ABSTRACT

Objective To construct recombinant lentiviral vector of microRNA-34a and observe the cell viability,cell cycle and apoptosis of hepatocellular carcinoma cells transfected with the vector system and treated with Doxorubicin.Methods Recombinant lentiviral vector containing microRNA-34a gene was constructed and transfected into 3 hepatocellular carcinoma cell lines,and cells were treated with Doxorubicin.The expression of microRNA-34a gene was detected by real-time PCR.The effect of microRNA-34a overexpression on hepatocellular carcinoma cells proliferation were quantified via MTT assay,cell cycle and apoptosis was evaluated by flow cytometry.Western blotting was used to evaluate the expression of cell cycle and apoptosis related protein.Results The successful construction of microRNA-34a recombinant lentiviral vector was confirmed by plasmid enzyme digestion and DNA sequencing.Compared with the control group,relative expression of microRNA-34a gene in hepatocellular carcinoma cells significantly increased ((HepG2:t=15.36,P＜0.01;Hep3B:t=36.75,P＜0.01;Bel-7402:t=24.17,P＜0.01)).Cells viability decreased (HepG2:t =7.12,P ＜ 0.01;Hep3B:t =8.89,P ＜ 0.01;Bel-7402:t =13.62,P ＜0.01),G1 phase cells increased significantly(HepG2:F =137.65,P ＜ 0.01;Hep3B:F =143.39,P ＜0.01;Bel-7402:F =1 306.47,P ＜ 0.01) and cell apoptosis increased(HepG2:F =386.14,P ＜ 0.01;Hep3B:F =881.94,P ＜ 0.01;Bel-7402:F =885.89,P ＜ 0.01).Conclusions MicroRNA-34a recombinant lentiviral vector (LV-hsa-mir-34a) transfected hepatocellular carcinoma cells overexpress microRNA-34a,reduce the malignant biological behavior.MicroRNA-34a recombinant lentiviral vector (LV-hsa-mir-34a) enhance the in vitro inhibitory effects of Doxorubicin on hepatocellular carcinoma cell lines.