EFFECT OF RNA INTERFERENCE TARGETING AT SURVIVIN GENE ON APOPTOSIS AND CHEMOTHERAPY OF HEPATOCELLULAR CARCINOMA CELL LINE HepG2

Objective To investigate the effect of RNA interference targeting at survivin gene on the apoptosis and chemosensitivity of hepatocellular carcinoma cell line HepG2. Methods Survivin gene sequence specific shRNA vectors pshRNA-survivin-387 were constructed.HepG2 cells cultured in 6-well culture dish were divided into five groups: blank control group(without any drug),negative control group(3.2 μg pshRNA-survivin-NC 3/well),low-dose group(0.8 μg pshRNA-survivin-387/well),medium-dose group(1.6 μg pshRNA-survivin-387/well) and high-dose group(3.2 μg pshRNA-survivin-387/well).After 48 h of transfection,cultured cells were harvested,and the apoptotic index was examined by flow cytometry.HepG2 cells cultured in 96-well culture dish were divided into three groups:blank control group(without any drug),negative control group(0.2 μg pshRNA-survivin-NC/well),and pshRNA-survivin-387 group(0.2 μg pshRNA-survivin-387/well).MTT assay was used to detect the rate of cellular inhibition effected by C-DDP(2.0 mg/L) after 24,36,48 and 60 hours. Results The apoptotic index of the low-,medium-and high-dose pshRNA-survivin-387 groups were significantly higher than those of the blank control group(F=228.87,q=14.21-38.21,P0.01) and those of the negative control group(q=10.45-34.40,P0.01).Apoptotic index of high-dose pshRNA-survivin-387 groups were significantly higher than that of the low-dose pshRNA-survivin-387 groups(q=23.99,P0.01).The rate of inhibition of C-DDP in pshRNA-survivin-387 groups were significantly higher than those of the control groups(F=235.88-910.86,q=28.04-52.28,P0.01). Conclusion Transfection of the pshRNA-survivin-387 to HepG2 cell can induce the apoptosis of liver cancer cells,and increase the sensitivity of the cells to chemotherapy.