Objective:To investigate the possibility of survivin inhibition by lentiviral vector-mediated RNA interference and the influence on cell apoptosis in pancreatic cancer cell line.
Methods:The lentiviral vector of SiRNA targeted against survivin(LV-shRNA-survivin-1, LV-shRNA-survivin-2, LV-shRNA-survivin-3) was constructed and transfected into the packaging cells 293T, and then the supernatant with virus was collected to transfect SW1990 cells. Quantitative real-time fluorescent PCR and Western-blot were used to detect the expression of survivin. DAPI staining and detection of enzymatic activity of caspase 3/7 were employed to examine cell apoptosis.
Results:Three lentiviral vector-survivin-shRNA were constructed successfully. In the LV-shRNA-survivin-1 group,the survivin mRNA and protein expression inhibitory rate was 73.50% and 87.64% respectively; when compared to control group, the activity of caspase-3/7 increased significantly,which showed a 14.5-fold increase, and apoptosis increased 11.95%.
Conclusions:Lentiviral vector-mediad RNA interference targeted against survivin can effectively inhibit survivin expression and increase cell apoptosis significantly.