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

作　　者：刘艳红 李静 王娜[2] 张骥[2] 李坤[2] LIU Yanhong;LI Jing;WANG Na;ZHANG Ji;LI Kun(Experiment Teaching Center of Basic Chemistry,Sichuan University,Chengdu 610064,China;College of Chemistry,Sichuan University,Chengdu 610064,China)

机构地区：[1]四川大学基础化学实验教学中心,成都610064 [2]四川大学化学学院,成都610064

出　　处：《实验室研究与探索》2023年第9期146-150,共5页Research and Exploration In Laboratory

基　　金：国家自然科学基金项目(32071276);四川大学新世纪高等教育教学改革工程研究项目(SCU10259);四川大学研究生教育教学改革研究项目(GSSCU2021093)。

摘　　要：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.

关 键 词：基因沉默 小干扰RNA 基因传输 细胞凋亡 

分 类 号：G642.423[文化科学—高等教育学]