关键词: RNA干扰, Survivin, hTERT, 结肠癌

Abstract: Objective To investigate the Influence of Survivin and hTERT gene on cell proliferation and apoptosis in human colorectal carcinoma cell line SW480 and to provide experiment evidence for gene therapy of colorectal carcinoma. Methods Plasmids carrying shRNAs targeting survivin and hTERT were designed, constructed and transfected into SW480 cells. SW480 cells were divided into blank group, blank Plasmid control group, survivin RNAi group, hTERT RNAi group and Survivin-hTERT RNAi group. The telomerase activity was examined by TRAP-PCR-ELISA analysis 48h after hTERT-shRNA transfection．Survivin and hTERT mRNA and protein expression was analyzed by RT-PCR and Western blot. Cell apoptosis, proliferation were measured by flow cytometry, cck-8 assay. Results Telomerase activity in SW480 cells of Survivin-hTERT RNAi groups were significantly decreased compared with the blank group（P<0.01）. The expression of survivin and hTERT mRNA, proteins in the Survivin-hTERT RNAi group was reduced by 82.8% and 73.6%（P<0.01），79.2% and 66.7% （P<0.01）respectively. The inhibitory rate of cell proliferation of Survivin-hTERT RNAi group was 43.6%±0.1%（P<0.01）. The apoptosis rate was 39.2%±2.3%（P<0.01） in the Survivin-hTERT RNAi group. Conclusion the Survivin-hTERT RNAi group could significantly reduce the protein expression of survivin and hTERT mRNA, inhibit cell proliferation, induce cell apoptosis in human colorectal carcinoma cell line SW480.

Key words: RNA interference, Survivin, hTERT, Colorectal carcinoma