Research of inhibition of survivin in rat HSC-T6 cell by siRNA interference

Abstract

OBJECTIVE: To explore the effects and mechanisms of siRNA targeting survivin of inducing apoptosis in rat HSC-T6 cells. METHODS: The experiment was divided into blank group, pGPU6/GFP/Neo-shNC group and pGPU6/GFP/ Neo-siRNA group. The siRNA was transfected into HSC-T6 cells mediated by Lipofectamine TM2000 for 24 h, and then the effi ciency of transfection was observed by fl uorescence microscopy. After transfection for 48h, the expression of survivin mRNA and protein was assessed by reverse transcription-polyme-rase chain reaction (RT-PCR) method and Western-blot, and the form of cells was observed by microscopy. The apoptosis rate of HSC-T6 cells was measured by the fl ow cytometry with PI staining. The expression of caspase-3 protein was assessed by western-blot. RESULTS: The prominent apoptosis of the pGPU6/GFP/Neo-siRNA group by PI staining was high, there was signifi cant difference comparing with blank group and pGPU6/GFP/Neo-shNC group (p < 0.05). The prominent apoptosis of blank group and pGPU6/GFP/Neo-shNC group was weak, there was no signifi cant difference among blank group and pGPU6/GFP/Neo-shNC group (p > 0.05). The expression of caspase-3 by Westernblot in pGPU6/GFP/Neo-siRNA group was high, there was signifi cant difference comparing with blank group and pGPU6/GFP/Neo-shNC group (p < 0.05), the expression of caspase-3 in blank group and pGPU6/GFP/ Neo-shNC group was weak, there was no signifi cant difference among blank group and pGPU6/GFP/NeoshNC group (p > 0.05). CONCLUSION: siRNA targeting survivin can inhibit the expression of survivin mRNA and protein in rat HSC-T6 cells. Expression of survivin is negative correlation with expression of caspase-3. siRNA targeting survivin may upregulate expression of caspase-3 and increase apoptosis of rat Ref. 24). Text in PDF www.elis.sk.