Construction Of A Vector And The Efficacy Evaluation Of Inhibition For RNA Interference Targeting VEGF Gene Of HepG-2

Objective: There are many reasons for malignant proliferation of hepato- carcinoma.Vascular endothelial growth factor(VEGF)has been implicated as a critical molecular signal in tumour development,by promoting angiogenesis and possibly by exerting autocrine functions in tumour cells.Some studies showed that VEGF was over expressed in hepatocarcinoma.This paper attempts to investigate the inhibitory effect of retroviral vector-based RNA interference(RNAi) on the expression of vascular endothelial growth factor(VEGF) protein in HepG-2 cells and provide an experimental basis for cure hepatocarcinoma with RNAi methods.Methods: The pSUPER.retro.neo-VEGF-siRNA expression vector was constructed by gene recombination. The retroviral vector which expresses the specific small hairpin RNA targeting VEGF gene was transfected into hepatoma cells(HepG-2) by Lipofectamine 2000 and positive colones were screened by G418. The VEGF expression of HepG-2 cells before and after transfection were detected by RT-PCR and Western-blot methods.Results:pSUPER.retro.neo-VEGF-siRNA expression vector was successfully constructed. The expressions of VEGFmRNA and protein were decreased markedly after pSUPER.retro.neo-VE??iRNA transfected using Lipofectamine2000 and the inhibitive ratio was 65%and 64% respectively.Conclusion:The research provided an experimental basis for cure hepatocarcinoma with RNAi methods.