Effect Of SiRNA Silencing Survivin Gene On Apoptosis Of Nasopharyngeal Carcinoma CNE-2 Cells

Objective Synthesis of Survivin small interfering RNA(siRNA)sequence and transfection of human undifferentiated nasopharyngeal carcinoma cell line CNE2,observed the situation of CNE-2 cells which transfected with Survivin siRNA,including Survivin mRNA and protein expression in vitro.CNE-2 cells were transfected with high efficiency siRNA,and the apoptosis of CNE-2 cells were detected.Methods According to the gene bank of Survivin sequence,design and synthesis of 3 segments specific Survivin siRNA,then siRNA transfected with CNE-2 cell line,set the blank control and negative control group,the Survivin mRNA of CNE-2 cells were detected by RT-qPCR,and the relative expression of mRNA were calculated by 2-AACt method.The expression of Survivin protein were detected by Western blot,and the apoptosis of CNE-2 cells were detected by flow cytometry and in situ apoptosis.Results Each group of siRNA was transfected into CNE-2 cells and compared with the Survivin mRNA relative expression of the control group,the Survivin mRNA relative expression of the transfected siRNA1,2,3 sequence group,was 0.320± 0.061(p<0.01),0.697±0.136(p<0.05),0.517±0.013(p<0.01),respectively,and the difference was statistically significant.Compared with the transfection of nonsense sequence group,the inhibitory rates of mRNA expression in siRNAl and siRNA3 groups were(68.46 ± 7.94)%,(49.44± 3.78)%and siRNA2 group was not statistically significant.The relative expression of the protein was only reduced in the siRNA group and the expression inhibition rate was(30.61±2.47)%.Flow cytometry was used to detect the apoptotic rate of transfected siRNA1,nonsense sequences CNE-2 cells and untransfected cells,the apoptotic rates of each group were 26.14%,10.65%,0%,respectively,and the apoptotic rate of transfected siRNAl group was 15.49%higher than that of the transfected nonsense group.TUNEL method was used to detect CNE-2 cells,and most of the nuclei of transfected siRNA1 group were brown,and the nuclei of the transfected nonsense and non transfected cells were blue,the percentage of apoptosis was increased.Conclusion SiRNA interference silencing Survivin gene can effectively inhibit the proliferation and induce apoptosis of CNE-2 cells.This study provides in vitro experimental support for which Survivin gene may serve as a target for the treatment of nasopharyngeal carcinoma and Survivin gene silencing may an effective way for the treatment of nasopharyngeal carcinoma.