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1、siRNA诱发K562／Adr细胞凋亡的机制研究?232?ModemPracticalMedicine,April2006.Vo1.1,.siRNA诱发K562/Adr细胞凋亡的机制研究冯敏华,张我,顾静文,林果为球【摘要】目的研究siRNA引起K562/Adr细胞凋亡的机制.方法siRNA重组质粒pSilenceMDR-1和pSilenceneo-GSTTr转染K562/Adr细胞,透射电镜观察K562/Adr细胞超微结构变化;Westernblot检测凋亡相关蛋白Bcl-2的改变.结果透射电镜下,pSilenceMDR
2、一1和pSilencenee—GSTxr转染组细胞胞浆浓缩,线粒体出现肿胀空泡样变,胞核内染色质浓集,染色质形成新月体样改变并有凋亡小体形成等典型的凋亡改变;Westernblot结果表明pSileneeMDR?l和pSilenceneo-GST~r转染组发生凋亡的K562/Adr细胞中,Bcl-2含量明显减少,与mock相比差异具有显着性意义(P<0.05).结论siRNA重组质粒pSilenceMDR-1和pSilenceneo-GSTxr通过抑制Bcl-2表达诱导K562/Adr细胞发生凋亡,逆转其多药耐药的发
3、生,从而为siRNA逆转白血病多药耐药提供坚实的理论依据.【关键词】siRNA;pSilenceMDR-1和pSilenceneo—GST'rr;细胞凋亡;基因,bclI2【中图分类号】R733.7【文献标识码】A【文章编号】1671—0800(2006)04—0232—04ApoptoticmechanismofcelllineK562/AdrinducedbySiRNAexpressionvectorFENGMin?Hua,ZHANGTao.GUJing一叫en,eta1.(HuazhanHospitalAO~liat
4、edFudanUniversity.Shanghai2000400.China)【Abstract】ObjectiveTostudytheapoptosismechanismofcelllinesK562/AdrinducedbysiRNA.MethodspSilenceMDR-landpsilencenee-GST盯weretransfectedintoK562/Adr.theuhrastmeturalchangesofK562/Adrcelllinewereobservedbytransmissionelectronm
5、icroscope-tileamountofbcl-2wasexaminedbyWesternblot.ResultsUndertheelectronmicroscope.theeell'SnucleustransfectedbypSilenceMDR.1andpSilenceneo-GSTwbecameshrinking.thecrescentandapoptoticcarpusclewerefoundinchromatin.andthereweremorevacuolasintheconcentratedcytopla
6、sm.AftertransfectedwithpSilence-MDRIandneeGSTTr,thea.mountofbcl-2wasdecreasedfrom0.754-0.03to0.524-0.12,whichWassignificantlydifferentfromcentrelgroup(P<0.05).ConclusionpSilence-MDR1andpSilence.GSTTrcaninducecelllineK562/Adrapoptosisbydecreasingbcl-2.reversethe
7、multidrugresistanceofK562/Adrcellline.andprovidethetheoreticevidenceforreversingmultidrugresistanceofleukemiacellbysiRNA.【KeyWords】siRNA;pSilenceMDR1.pSilencenee—GSTTr;Apoptosis:Genes;bel,2[ModernPracticalMedicine.2006.18(4):232—234.238]白血病化疗失败的主要原因是白血病细胞的多药耐药性(MD
8、R),主要涉及多药耐药基因(MDRI),耐药酶(GSTrr)等多种机制.近年来的研究表明,一些小的干扰RNA(smallinterferingRNA,siRNA)可以高效,特异地阻断体内靶基因的mRNA,使其降解,发生转录后基因沉默,称为RNA干扰(RNAinterfering,RNAi).本实验利用成
