The TRAIL On Expression Of HepG2 Cells And The Study Of Apoptosis Mechanism

Objective:Plasmid stability and the high expression of Hep G2 cells in the hepatocellular carcinoma cell line by lentivirus carrying TRAIL gene,Detectionthe expression of protein and proliferation of Hep G2 cells by TRAIL and Observation of TRAIL-Hep G2 cells and the effect of chemotherapy drugs used in combination by the method of MTT,The last to explore the possible mechanism of the apoptosis.Methods:1.Hepatoma cell line Hep G2 cell cultures2.Construction recombinant lentiviral vector p CDH-CMV-TRAIL-EF1-GFP-T2A-Puro;3.Lentivirus infected cell protein expression of TRAIL Hep G24. MTT detection of cell growth inhibition rate5.Hep G2 cell apoptosis and cell cycle conditions by flow cytometryResults:Restriction end nuclease analysis and DNA sequencingconfirmedthe recombinant lentiviral vector carrying TRAIL, and when MOI =40 found that the recombinant lentiviral transfected efficient in vitro,the stable high expressions of TRAIL protein were confirmed by Western blotting.The MTT assay detected stably transfected with TRAIL-HG2 + impact of drugs on cell proliferative activity of cells was significantly lower than Hep G2 and TRAIL-Hep G2 group(P <0.05), After the application of flow cytometry examination Hep G2 cell apoptosis and cell cycle, apoptosis was found in the experimental group was significantly higher, and the p-value <0.05,have the statistically significance.Conclusion:Above all that the Lentiviral vector has been constructed successfully with carrying TRAIL gene and realizes the stable and high expression in Hep G2 cells.Through the application in the control group and the experiment group with the chemotherapy of 5-FU in 0.1 g/m L,we can get the following results that, The apoptosis ratio betwween two groups was statistically significant difference(P<0.05). Further show that the drug can enhanced apoptosis mediated in TRAIL Hep G2 cells, and can significantly inhibit the growth and reproduction of hepatocellular carcinoma cells.