VEGF-siRNA expression construction vector and its inhibition of vascular endothelial growth factor in Tca8113 cells

Objective To construct the vector carrying siRNA of human vascular endothelial growth factor(VEGF) and to evaluate its inhibitory effect on VEGF in Tca8113 cells.Methods Sequencer of short hairpin RNA targeting VEGF of Tca8113 cells was designed and synthesized.Psilencer 2.1-U6-neo-VEGF-siRNA(Pu-VEGF-siRNA) was constructed and transfected to Tca8113 cells by lipofectamine 2000.Eukaryotic expression vector(Pu-HK) as the experiment control group,non-transfection cell was used as negative control group,all of which were transfected to Tca8113 cells by lipofectamine 2000.The expression of VEGF protein in Tca8113 cells transfected with Pu-VEGF-siRNA was detected by immunohistochemistry and enzyme linked immunosorbent assay(ELISA) respectively.Results After evaluation and sequencing,Pu-VEGF-siRNA was successfully constructed.Compared to the negative and experiment control groups,the expression of VEGF protein were significantly decreased in the Pu-VEGF-siRNA group(P 0.05);but there was no significant difference between the two control groups(P 0.05).Conclusion Pu-VEGF-siRNA expression vector is successfully constructed and it can significantly inhibit VEGF gene expression in Tca8113 cells.