siRNA抑制K562／ADM细胞mdr1基因表达并逆转其耐药性.docVIP

siRNA抑制K562／ADM细胞mdr1基因表达并逆转其耐药性 58 《中国癌症杂志》2007年第l7卷第1期 CH[NAONCOLOGY2007Vo1.17No.1 siRNA抑靠I】K562/ADM细胞mdrl 基因表达并逆转其耐药性 [摘要]背景与目的:白血病耐药性是白血病治疗中的难点,RNAi技术具有特异,高效,毒性小的特点,可高 效,特异地抑制特定基因的过度表达.本文研究小干扰RNA分子(siRNA)对白血病多药耐药K562/ADM细胞 mdrl基因表达和耐药性的影响.方法:设计,筛选和合成针对mdrl基因的siRNAs(si—mdrl—l,si—mdrl一2),脂质体 介导转染K562/ADM细胞;RT—PCR法检测mdrlmRNA的转录;流式细胞术测定P一糖蛋白(P—gP)表达水平;MrI丫r 法检测K562/ADM细胞对多柔比星(阿霉素,ADM)的敏感性.结果:si—mdrl一1,si—mdrl-2转染24h和48h,si—mdrl一1 的抑制率分别为55.5%和22.5%,而si—mdrl一2则分别为16.0%和57.6%.si—mdrl—l和si—mdrl一2作用72h时, P—gp的表达强度分别下降74%和85%.si—mdrl—l和si—mdrl一2均可提高K562/ADM细胞对多柔比星的敏感性,逆 转其耐药性,逆转倍数分别为2.52倍和1.96倍.结论:siRNA可特异性地沉默mdrl基因的表达,逆转P—gP介导 的白血病细胞耐药性. [关键词]siRNA;mdrl基因;白血病;耐药性 中图分类号:R73—362;R733.7文献标识码:A文章编号:1007—3639(2007)0l-0058-04 siRNAssilenceexpressionofmdrlgeneanditsroleinreversingdrug—resistanceinl62/ADM cellsGAOLi-pingWElHu—lai,jlNGTao,WUYong-fie,CHENJing,sUNjingYIJuan,zHAo Huai.shun(LaboratoryCenterforMedicalScience.LanzhouUniversity;KeyLaboratoryofPreclinicalStud. YforNewDrugsofGansuProvince,Lanzhou,Gansu73000,ChinaJ Correspondeneeto:WEIHu—tE—mail:weihulai@hu.edu.cn 【Abstract】Backgroundandpurpose:Drug—resistanceisthemainobstacleintermsofefficacyofchemotherapyfor leukemia,RNAinterference(RNAi)strategypossessesthecharacteristicsofspecilization,high—efficiencyandlow—toxicity, andcaneffectivelyandspecificallyinhibittheoverexpressionofgivengene.Thisstudywasdesignedtoinvestigatethe effectofsmallinterferingRNA(siRNA)onexpressionofmdrlgeneanddrug—resistanceinmuhidrug—resistanthumanleu— kemiaK562/ADMcel1.Methods:Humanmuhidrug.resistantleukemiacelllineK562/ADMover—expressingmdrlgenewas usedasthetargetcells,TwosiRNAs(si—mdrl—landsi—mdrl一2)targetedmdrlgenewerechemicallysynthesizedandtrans. fectedintoK562/ADMcells.ExpressionofmdrlmRNAwasdeterminedbyRT—PCR.P—glycoprotein(P—gP)expression wasmeasuredusingflowcytometry(FCM).andthesensitivityofK562/ADMcellstoadriamvcinwasassessedwithaMqq eolorimetricassay.Results:TwosiRNAsfsi—mdrl—landsi—mdrl一21speciallydesignedinthisstudycouldmarkedlydown r