关键词: RNA干扰, Survivin, hTERT, Sp1

Abstract: Objective Observation of Survivin gene interference effect on the proliferation of SW480 cells, to investigate the effects of the gene expression of hTERT and its mechanism of interference. Methods Small hairpin RNA targeting Survivin was synthesized and recombinant plasmid pGenesil-1.1-Survivin was constructed. The siRNA recombinant was transfected into SW480 cell lines. These cell proliferation activities were detected by MTT and the cell cycles were analyzed by flow cytometry. The expression levels of hTERT and Sp1 mRNA and protein were analyzed by The real-time fluorescence quantitative PCR and western blotting, respectively. Telomerase activity was examined by TRAP-ELISA analysis. To construct the interference vector targeting Sp1 and transfect into SW480 cell, hTERT mRNA and protein, telomerase activity were examined. Results The cell proliferation rate of SW480 cells transfected with siRNA- Survivin recombinant vector was significantly decreased, the inhibitory rate of cell proliferation was 34.44%(P<0.05)，an increase in the percentage of cells in the G0 / G1 phase and a decrease in the percentage of cells in the S and G2/M phase(P<0.01)．that expressions of hTERT and Sp1 mRNA and protein, telomerase activity were decreased in shRNA-Survivin and shRNA-Sp1 transfected cells, respectively(P<0.01). Conclusion silencing Survivin gene significant inhibition the proliferation of SW480 cells and down-regulation of hTERT gene expression, its mechanism may be related with down-regulation of Sp1 gene expression.

Key words: RNA interference, Survivin, hTERT, Sp1