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AIM:To inhibit the expression of vascular endothelial growth factor(VEGF)in colon cancer cell line by RNA interference(RNAi). METHODS:Followed the service of E-RNAi,we designed and constructed two kinds of shRNA expression vectors aiming at the VEGF gene,then transfected them into colon cancer HT29 cells by lipofectamine~(TM)2000.The level of VEGF mRNA was investigated by RT-PCR and Northern blotting.The protein expression of VEGF was observed by immunofluoresence staining and Western blotting. RESULTS:We got two kinds of VEGF specific shRNA expression vectors which could efficiently inhibit the expression of VEGF in HT29 cells.RT-PCR,Northern blotting,immunofluoresence staining and Western blotting showed that inhibition rate for VEGF expression was up to 42%,89%,73% and 82%,respectively. CONCLUSION:The expression of VEGF can be inhibited by RNA interference in HT29 cells.
AIM: To inhibit the expression of vascular endothelial growth factor (VEGF) in colon cancer cell line by RNA interference (RNAi). METHODS: Followed the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors aiming at the VEGF gene, then transfected them into colon cancer HT29 cells by lipofectamine ~ (TM) 2000. The level of VEGF mRNA was investigated by RT-PCR and Northern blotting. The protein expression of VEGF was observed by immunofluoresence staining and Western blotting. RESULTS: We got two kinds of VEGF specific shRNA expression vectors which could highly inhibit the expression of VEGF in HT29 cells. RT-PCR, Northern blotting, immunofluoresence staining and Western blot showed that inhibition rate for VEGF expression was up to 42%, 89%, 73 % and 82%, respectively. CONCLUSION: The expression of VEGF can be inhibited by RNA interference in HT29 cells.