沉默ERCC1基因对卵巢癌细胞A2780顺铂耐药性的影响.docx

沉默ERCC1基因对卵巢癌细胞A2780顺铂耐药性的影响

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作者：孙业红，陈富强，朱莹，李体远

【摘要】目的：研究RNA干扰切除修复交叉互补基因1(ERCC1)的表达后，卵巢癌细胞A2780对顺铂耐药性的变化。方法：A2780细胞分别转染ERCC1基因的siRNA和GFPsiRNA,并给予顺铂(使其终浓度达到3μmol/L),蛋白质印迹法分别检测顺铂给药前后ERCC1蛋白的表达，并用MTT法检测顺铂不同浓度下A2780细胞的存活率。结果：①转染ERCC1siRNA后，实验组蛋白质表达量较对照组显著降低；②ERCC1siRNA+顺铂组的ERCC1蛋白表达要弱于ERCC1siRNA+PBS组，而GFPsiRNA+顺铂组的蛋白表达要弱于GFPsiRNA+PBS组；③顺铂(浓度在0.75～12μmol/L之间)呈浓度依赖性地降低A2780细胞的存活率；④在相同浓度顺铂的作用下，与GFPsiRNA组相比，ERCC1siRNA组的顺铂量效曲线左移。结论：顺铂能抑制沉默ERCC1基因后A2780细胞的生长，且在一定浓度范围内呈浓度依赖性；ERCC1是参与顺铂耐药性产生的重要

因素之一。

【关键词】RNA干扰；切除修复交叉互补基因1;顺铂；耐药性；卵巢癌

[Abstract]Objective:ToinvestigatetheinfluenceofRNAinterferingtheexpressionofexcisionrepaircrosscomplementing1(ERCC1)geneonthesensitivityofovariancancercellA2780toCisplatin.Methods:SiRNAandGFPsiRNAwastransfectedtoknockoutERCC1geneinstantaneousandmakeacontrolmannerrespectively.Cisplatin(finalconcentrationof3μmol/L)wasgiventotheknockoutedcells.WesternblotwastakenfortheanalysesofERCC1proteinexpressioninA2780cellsbeforeandafterCisplatinwasgiven.Methylthiazolyltetrazolium(MTT)colorimetricassaywereusedtodeterminethelivabilityofthecancercellsindifferentCisplatinconcentration.Results:①AftertransfectionofERCC1siRNA,TheresultsshowedtheERCC1proteinexpressionlevelofcontrolgroupwassignificantlyhigherthanthatofexperimentalgroup;②ERCC1siRNA+CisplatingroupandGFPsiRNA+CisplatingroupexpressedlessERCC1proteinthanERCC1siRNA+PBSgroupandGFPsiRNA+PBSgrouprespectively;③Cisplatin(attheconcentrationof0.75～12μmol/L)inhibitedthelivabilityofA2780cellsinadosedependentmannerindespiteofsiRNA;④Atasameconcentrationof

Cisplatin,thedoseeffectcurveofknockoutgroupwaslefttothatofcontrolgroup.C