siRNA逆转白血病K562/ADM细胞对抗癌药物耐受性的研究  

Reversal of drug-resistance of leukemia K562/ADM cells to chemotherapeutical agents by small interfering RNA

作　　者：祁萍[1] 安娴[1] 高丽萍[1] 魏虎来[1] 

机构地区：[1]兰州大学医学实验中心甘肃省新药临床前研究重点实验室,甘肃兰州730000

出　　处：《中华肿瘤防治杂志》2008年第9期648-651,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30370607);甘肃省新药临床前研究重点实验室开放基金项目(GSKFKT-0502)

摘　　要：目的:研究小干扰RNA(smallinterfering RNA,siRNA)对多药耐药细胞K562/ADM耐药性的逆转效应。方法:以K562/ADM细胞为靶细胞,针对mdr1基因mRNA不同靶点设计合成4对siRNA分子,脂质体介导转染K562/ADM细胞,RT-PCR和实时荧光定量PCR检测mdr1基因mRNA的表达水平,流式细胞术(FCM)测定P-糖蛋白(P-gp)表达;MTT比色法检测K562/ADM细胞对多柔比星(ADM)、柔红霉素(DNR)和依托泊苷(Vp-16)的敏感性。结果:选择mdr1mRNA的4个靶点设计合成的siRNA中,针对333靶点的siRNA(simdr1/333)对mdr1基因的表达具有较好的沉默效果,RT-PCR和实时定量PCR检测,mdr1 mRNA表达分别降低69%和63%;FCM检测P-gp的表达强度(MFI)降低约50%。si-mdr1/333可提高K562/ADM细胞对ADM、DNR和Vp-16的敏感性,耐药逆转倍数分别为3.45、1.70和2.91倍。结论:siRNA能特异性地阻抑白血病多药耐药细胞K562/ADM细胞mdr1基因的表达,逆转其对抗癌药物的耐受性。OBJECTIVE:To explore the effect of small interfering RNA(siRNA) on reversing multidrug-resistant leukemia cells. METHODS: Human multidrug-resistant leukemia cell line K562/ADM overexpressing mdr1 gene was used as the target cells. Four siRNAs speciafically targeting mdr1 gene were chemically synthesized and transfected into K562/ADM cells with RNAi-Mate. The silencing efficiency of siRNAs were evaluated by the inhibition of mdr1 mRNA expression measured with RT-PCR and Real-time PCR,and the inhibition of P-...

关 键 词：RNA 小分子干扰 基因 mdr1 基因沉默 抗药性 肿瘤 白血病 

分 类 号：R733.7[医药卫生—肿瘤]