Survivin基因沉默诱导宫颈癌细胞凋亡的实验设计

Experimental Design of Survivin Gene Silencing to Induce Hela Cell Apoptosis

采用基因干扰的方法敲低Survivin蛋白表达,探究Survivin表达下调对人宫颈癌细胞凋亡的影响。实验将聚乙烯亚胺(PEI)作为siRNA传输的载体,在无血清条件下,聚乙烯亚胺有效压缩、保护和传输siRNA。PEI/siEGFP复合物高效转染Hela/EGFP细胞后,EGFP蛋白的表达量显著下降。而有血清条件下,PEI/siEGFP复合物不稳定,导致PEI/siEGFP的细胞摄取降低,影响了EGFP基因沉默的效果。Survivin基因干扰实验显示高效转染PEI/siSur复合物后,显著降低了Survivin基因表达。而Survivin表达的下调激活了凋亡信号通路中关键性激酶Caspase-3,从而诱导Hela细胞凋亡。该实验为宫颈癌的治疗提供了新的策略。

To explore the effect of down-regulation of Survivin protein expression on the apoptosis of human cervical cancer cells,a gene interference experiment was developed.Polyethylenimine(PEI)was used as a carrier for siRNA delivery.Under serum-free conditions,PEI can effectively condense,protect and deliver EGFP-siRNA.PEI/siEGFP polyplex exhibited high transfection efficiency in Hela/EGFP cells,the expression of EGFP protein is significantly reduced.However,PEI/siEGFP polyplex is unstable in the presence of serum,which results in its dramatical decrease of cellular uptake and gene-silencing efficiency.In addition,PEI/siSur polyplex shows efficiently down-regulated the expression of Survivin mRNA in Hela cells,causes the activation of caspase-3,a key kinase in the apoptosis signaling pathway and induces apoptosis of Hela cells.This work provides a promising strategy for the treatment of cervical cancer.