SiRNA targeting survivin enhances human osteosarcoma cell MG63 apoptosis

[Objective] To explore the effect of siRNA on the expression of survivin in gene and leading apoptosis in osteosarcoma cell line MG63 using siRNA eukaryotic expression vector. [Method] According to survivin cDNA coding sequence, the specific RNA interference (RNAi) fragments targeting survivin gene were designed and synthesized, which were cloned into pSilencer 3.0-H1 neo plasmid vector, and the siRNA eukaryotic expression vector siRNA survivin targeting survivin gene was constructed. After the vector was constructed, MG63 cells were transfected with negative control vector or RNAi vectors and selected by G418. Expression of protein of surviving in the stable transfected cells was investigated by Western blot. The above cells were cultured, then growth curve was drawn. These cells cultured on cover slips were observed through electron microscopy. Apoptosis analysis was finished by AnnexinⅤand Hoechst staining. [Results] The specific siRNA eukaryotic expression vector PsiS targeting survivin gene were constructed successfully. The stable transfectants containing negative control vector and PsiS were obtained. Expression of protein of survivin was inhibited significantly in MG63/PsiS cells, whereas survivin gene expression levels were hardly changed in other groups. Otherwise, apoptosic rate was increased significantly in MG63/PsiS cells than other two group (P 0.01). [Conclusion] Survivin gene expression can be suppressed markedly by specific siRNA which induced apoptosis in MG63 cells, the current results establish the experimental foundation for further studying the biological functions and its mechanisms of survivin in MG63 cells.