中国生物工程杂志

Objective To inhibit the expression of Vascular Endothelial Growth Factor (VEGF) by RNA interference in HT29 cells. Methods Using the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors which were aimed at the VEGF gene, then transfected them into colon cancer HT29 cells by lipofectamineTM 2000. The level of VEGF mRNA was determined by RT-PCR and Northern blotting. The protein expression of VEGF was examined by Immunofluoresence staining and Western blotting. Results The two kinds of VEGF specific shRNA expression were found to efficiently inhibit the expression of VEGF in HT29 cells, the inhibition rate showed by RT-PCR, Northern blotting, Immunofluoresence staining and Western vectors blotting were 42%, 88%,73% and 82%, respectively. Conclusion The expression of VEGF could be inhibited by RNA interference in HT29 cells.