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 LipofectamineTM2000 for 24 h, and then the efficiency of transfection was observed by fluorescence microscopy. After transfection for 48h, the expression of survivin mRNA and protein was assessed by reverse transcription-polymerase 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 flow 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 significant difference comparing with blank group and pGPU6/GFP/Neo- shNC group (p<0.05). The expression of caspase-3 by Western-blot in pGPU6/GFP/Neo-siRNA group was high, there was significant difference comparing with blank group and pGPU6/GFP/Neo-shNC 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 up-regulate expression of caspase-3 and increase apoptosis of rat HST-T6 (Fig. 6, Ref. 24).
Keywords: apoptosis.; caspase-3; hepatic stellate cell; siRNA; survivin.