The Molecular Mechanisms Are Involved In The Induction Of Apoptosis Of Human Hepatocellular Carcinoma Cells Hepg₂ By PGL3-hTERTp-HSV-TK/GCV System

Objective:Gene therapy has become more and more popular in the treatment of tumor. The aim of this study was to observe the molecular mechanisms are involved in the induction of apoptosis of human hepatocellular carcinoma cells HepG2 by pGL3-hTERTp-HSV-TK/GCV system.Methods:1.We detected the activity of the HepG2 cells and the L-02 cells through Tunnel and the methyl thiazolyl tetrazolium (MTT) after they were transducted by the recombinant adenovirus and adding GCV of different concentrations.2. The changes of cleavage products of Caspase-8,Caspase-3,Caspase-10,Survivin were analyzed by Western blot after the pGL3-hTERTp-HSV-TK/GCV system treatment of HepG2 cells.3. The inhibitors for Caspase-8, Caspase-3 and Caspase-10 were used to block the apoptotic pathway of HepG2 cells by pGL3-hTERTp-HSV-TK/GCV system.Results:1.pGL3-hTERTp-HSV-TK/GCV system targeting hepatocellular carcinoma cell killing effect, while the normal liver cells almost unaffected.2.Western blotting showed that with time, Caspase-8, Caspase-3 zymogen decreased, the activation segment increased, while Caspase-10 zymogen had no significant change.In addition, Survivin is gradually reduced.3.Caspase-8 inhibitor can inhibit significantly pGL3-hTERTp-HSV-TK/GCV system induced apoptosis in hepatoma cells, while the Caspase-3 inhibitor partially inhibited the apoptosis inhibitor. The inhibitory effect of Caspase-10 inhibitor is weak.Conclusion:PGL3-hTERTp-HSV-TK/GCV system induced apoptosis of HCC primarily activated death receptor pathway of apoptosis upstream of Caspase-8 caused, also involves other members of the caspase family, such as Caspase-3. However, for the upstream apoptosis signal Caspase-10 were not involved in the apoptotic process. In addition, involved in the activity of Survivin reduced.