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1、siRNA誘發(fā)K562／Adr細(xì)胞凋亡的機(jī)制研究?232?ModemPracticalMedicine,April2006.Vo1.1,.siRNA誘發(fā)K562/Adr細(xì)胞凋亡的機(jī)制研究馮敏華,張我,顧靜文,林果為球【摘要】目的研究siRNA引起K562/Adr細(xì)胞凋亡的機(jī)制.方法siRNA重組質(zhì)粒pSilenceMDR-1和pSilenceneo-GSTTr轉(zhuǎn)染K562/Adr細(xì)胞,透射電鏡觀察K562/Adr細(xì)胞超微結(jié)構(gòu)變化;Westernblot檢測(cè)凋亡相關(guān)蛋白Bcl-2的改變.結(jié)果透射電鏡下,pSilenceMDR
2、一1和pSilencenee—GSTxr轉(zhuǎn)染組細(xì)胞胞漿濃縮,線粒體出現(xiàn)腫脹空泡樣變,胞核內(nèi)染色質(zhì)濃集,染色質(zhì)形成新月體樣改變并有凋亡小體形成等典型的凋亡改變;Westernblot結(jié)果表明pSileneeMDR?l和pSilenceneo-GST~r轉(zhuǎn)染組發(fā)生凋亡的K562/Adr細(xì)胞中,Bcl-2含量明顯減少,與mock相比差異具有顯著性意義(P<0.05).結(jié)論siRNA重組質(zhì)粒pSilenceMDR-1和pSilenceneo-GSTxr通過(guò)抑制Bcl-2表達(dá)誘導(dǎo)K562/Adr細(xì)胞發(fā)生凋亡,逆轉(zhuǎn)其多藥耐藥的發(fā)
3、生,從而為siRNA逆轉(zhuǎn)白血病多藥耐藥提供堅(jiān)實(shí)的理論依據(jù).【關(guān)鍵詞】siRNA;pSilenceMDR-1和pSilenceneo—GST'rr;細(xì)胞凋亡;基因,bclI2【中圖分類號(hào)】R733.7【文獻(xiàn)標(biāo)識(shí)碼】A【文章編號(hào)】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]白血病化療失敗的主要原因是白血病細(xì)胞的多藥耐藥性(MD
8、R),主要涉及多藥耐藥基因(MDRI),耐藥酶(GSTrr)等多種機(jī)制.近年來(lái)的研究表明,一些小的干擾RNA(smallinterferingRNA,siRNA)可以高效,特異地阻斷體內(nèi)靶基因的mRNA,使其降解,發(fā)生轉(zhuǎn)錄后基因沉默,稱為RNA干擾(RNAinterfering,RNAi).本實(shí)驗(yàn)利用成
