Effects Of ShRNA Targeted Survivin And VEGF On Proliferation, Apoptosis And Angiogenesis For Human Pancreatic Cancer Cells

Objective:To investigate the effects of short hairpin RNA (shRNA)-targeted Survivin and/or VEGF inhibition on the proliferation, apoptosis of human pancreatic cancer cells Panc-1and human umbilical vein endothelial cells (HUVECs), which were seeded in the culture medium from Panc-1cells transfected with siRNA.Methods:Targeted small interfering RNA (siRNA) expression vectors of Survivin and VEGF were constructed and transfected into Panc-1cells. The downregulation of Survivin and VEGF expression was evaluated by Real-time PCR and Western blot analysis. The effects of targeted shRNA on the proliferation and apoptosis of Panc-1cells were analyzed by MTT assay and flow cytometry (FCM). The culture medium from Panc-1cells transfected with siRNA was collected and human umbilical vein endothelial cells (HUVECs) were seeded in this media. The proliferation and apoptosis of the HUVECs were also investigated by MTT assay and FCM.Results:1.Four vectors were designed, which included4Survivin-specific siRNAs designated as S1, S2, S3and S4and the expression of Survivin was downregulated. S4directed at Survivin were selected as the most effective inhibitors for investigation in the latter experiments. The cell viabilities of Panc-1cells and HUVECs in inhibition group were decreased compared with the controls. The cell apoptosis rates of Panc-1cells and HUVECs in inhibition groups were observed to be increased compared with the controls.2. Four vectors were designed, which included4VEGF-specific siRNAs designated as V1, V2, V3and V4and the expression of VEGF was downregulated. V3directed at VEGF were selected as the most effective inhibitors for investigation in the latter experiments. The cell viabilities of Panc-1cells and HUVECs in inhibition group were decreased compared with the controls. The cell apoptosis rates of Panc-1cells and HUVECs in inhibition groups were observed to be increased compared with the controls.3. The cell viabilities of Panc-1cells and HUVECs in the combined inhibition groups were markedly decreased compared with the controls. The cell apoptosis rates of Panc-1cells and HUVECs in the combined inhibition groups were observed to be significantly increased compared with the controls.Conclusion:1.The RNA interference-mediated downregulation of Survivin or VEGF expression inhibited proliferation and induced the apoptosis of human pancreatic cancer cells and inhibit tumor angiogenesis by inhibiting proliferation and inducing the apoptosis of human vascular endothelial cells.2. The simultaneous RNA interference-mediated downregulation of Survivin and VEGF expression inhibited proliferation and induced the apoptosis of human pancreatic cancer cells and inhibit tumor angiogenesis by inhibiting proliferation and inducing the apoptosis of human vascular endothelial cells.