RNA Interference Inhibited VEGF Expression And Mediated Gene Therapy Of Colon Cancer

Objective: To inhibit VEGF expression by RNA interference in vitro, and observe the apoptosis of Colon cancer cell when VEGF was knocked down, so that RNA interference could be used in clinical therapy.Methods: (1) Followed the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors which aimed VEGF gene, and verified by gene sequencing.(2) Transfected them into HEK293, colon cancer HT29, Hela and HepG2 cells by lipofectamineTM 2000 in vitro, the expression of VEGF was investigated by RT-PCR, Immunofluoresence staining,Northern Blotting and Western blotting. When the level of VEGF was knocked down, the apoptosis of tumor cells was investgated by Tunel,elctron microscope and Flow cytometer.Results: (1) We got two kinds of VEGF specific shRNA expression vectors, their gene sequence were correct. (2) We found they could efficiently inhibit the expression of VEGF in serveral cells lines showed by RT-PCR, Immunofluoresence staining,Northern Blotting and Western blotting, but the inhibition rates of VEGF were varied in different cells lines in vitro.When the level of VEGF was knocked down, Tunel,elctron microscope and Flow cytometer showed that the apoptosis of tumor cells were increased.Conclusion: The expression of VEGF could be inhibited by RNA interference, but differed with the change of cells lines, showed that RNA interference was cells lines dependent. The inhibition of VEGF could induce the apoptosis of tumor cells in vitro.