Effect Of RNA Interference Targeting Survivin Gene On The Apoptosis And Chemosensitivity Of Hepatocellular Carcinoma Cell Line HepG2

Objective:To observe the effects of RNA interference targeting survivn gene on the apoptosis and chemosensitivity of Hepatocellular carcinoma cell line HepG2and to investigate the feasibility of treatment on hepatoma with RNA interference targeting survivin gene.Methods:The survivin gene sequence specific shRNA vectors pshRNA-survivin-387 were constructed. Transfection of shRNA vectors was performed using the Lipofectmine2000 liposome. Then did the following indexes:(1)HepG2 cells were cultured in 6-well culture dish, and divided into 5 groups:blank control group(without any durg),negative control group (3.2ugpshRNA-survivin-NC/well), and low dose (0.8ug pshRNA-survivin-387/well), medium dose(1.6ug pshRNA-survivin-387/well) and high dose (3.2ug pshRNA-survivin-387/well) pshRNA-survivin-387 groups.After transfection for 48 h, cultured cells were harvested to be examined the Apoptotic index by flow cytometry.(2)HepG2 cells were cultured in 96-well culture dish, and divided into 3 groups:blank control group(without any durg), negative controlgroup (0.2ugpshRNA-survivin-NC/well), and pshRNA-survivin-387 groups (0.2ug pshRNA-survivin-387/well). MTT assay was used to detectcell rate of inhibition which were effected by C-DDP (2.0ug/ml) after 24h,36h,48h and 60h.Results:(1) The Apoptotic index of the low, medium and high dose pshRNA-survivin-387 groups were significantly higher than those of the other control groups(compared with blank control group, F=228.87,*q=14.21-38.21, p<0.01; compared with negative control group,#q=10.45-34.4, p< 0.01),and the Apoptotic index of high dose pshRNA-survivin-387 groups were significantly higher than the low dose pshRNA-survivin-387 groups((F=228.87, q=23.99, p<0.01).(2) The rate of inhibition of C-DDP in pshRNA-survivin-387 groups were significantly higher than those of other control groups(F=235.88-910.86, q=28.04-52.28,p<0.01).Conclusion:Transfection the pshRNA-survivin-387 to HepG2 cell can induce liver cancer cells apoptosis, sensitize liver cancer cells to chemotherapy.