Effect of inhibition of hPOT1 expression mediated by siRNA on Caspase-3 in HeLa cells / 中国药理学通报

ABSTRACT

Aim To inhibit the expression of human protection of telomere gene hPOT1 in HeLa cells using vector based on RNA interference(RNAi) technique and to investigate its effects on the activation of Caspase-3 in HeLa cell.Methods Three recombinant plasmids(our group constructed before) containing different hPOT1 target sequences(pmU6-shRNA1,pmU6-shRNA2 and pmU6-shRNA3)were transfected into HeLa cells by liposomal.The expression inhibition of hPOT1 was detected by RT-PCR and EMSA.The expression and activation of Caspase-3 were inspected by RT-PCR,Western blot analysis and Caspase-3 kit.Results After 48 hour′s transfecting,hPOT1 mRNA and protein level in HeLa cells reduced and the expression of Caspase-3 mRNA had no change.Caspase-3 zymogen decreased but its activity increased.Conclusions hPOT1 expression in HeLa cells can be significantly inhibited by using siRNAvectored RNAi and the down-regulation of hPOT1 expression can activate Caspase-3.