| ObjectiveTo investigate silencing of survivin by siRNA and its effect on the induction of apoptosis and the expression of ER stress protein in liver cancer cells.MethodsLipotransfect HepG2 cells with psiRNA-survivin1 or psiRNA-survivin 2 or psiRNA-survivin1+2. The transfection efficiency of experiment group and control group was examined by FCM and inverse fluoresence microscope. Detect the mRNA level of survivin and ER stress protein within HepG2 cells via Real Time PCR.Monitor the apoptosis-inducing effects of the survivin silencing with inverse microscope and flow cytometry after Annexin-V/PI double-labeling.ResultsThe results of experiments showed that the mRNA level of survivin in HepG2 transfected with psiRNA-survivin1/2/1+2 was significantly decreased compared to that of HepG2 transfected with pgenesil8-HK (irrelevent plasmid) or without transfection, while no significant difference was seen between the mRNA level of survivin in HepG2 transfected with pgenesil8-HK and that of non-transfected HepG2. Moreover, it was found the occurrence of apoptosis increased remarkbly in HepG2 following transfection with psiRNA-survivin1/2/1+2, and psiRNA-survivin1+2 silencing expression of survivin and inducing apoptosis is the most significant. In addition, the mRNA level of ER stress protein in HepG2 transfected with psiRNA-survivin 1/2/1+2 was significantly increased compared to that of HepG2 transfected with pgenesil8-HK or without transfection, while mRNA level of ER stress protein in HepG2 transfected with pgenesil8-HK and non-transfected HepG2 did not display Statistical difference.Conclusion: siRNA can silence the expression of survivin in liver cancer HepG2 cells, induces the apoptosis of HepG2,and it proves that psiRNA-survivin1+2 silencing expression of survivin and inducing apoptosis is the best; further study show that after survivin silencing by siRNA, it can also increases ER stress protein gene expression, its mechanism and significance remains to be explored. |
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