小分子干扰RNA对卵巢癌紫杉醇耐药株的多药耐药性逆转的研究  

Reversal of multidrug resistance in A2780/Taxol cells by small interference RNA of MDR1 gene.

作　　者：肖兰[1] 卢实[1] 高瑞[1] 梁铭霖[1] 刘福安[1] 王泽华[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院妇产科,湖北武汉430022

出　　处：《中国实用妇科与产科杂志》2007年第1期41-43,共3页Chinese Journal of Practical Gynecology and Obstetrics

摘　　要：目的探讨小分子干扰RNA(smallinterferingRNAs,siRNA)对A2780/Taxol细胞多药耐药性的逆转作用。方法2005-07-2005-12华中科技大学同济医学院附属协和医院根据多药耐药基因1(MDR1)序列设计2条siRNA,将siRNA包装入质粒,经转化、克隆、扩增、纯化,与pEGFPN-1质粒一起共转染A2780/Taxol细胞,流式细胞仪、MTT、RT-PCR和Westernblot分别检测转染效率、紫杉醇的半数抑制浓度(IC50)、细胞P-gp蛋白及MDR1mRNA表达。结果两种重组质粒与EGFP质粒转染效率无明显差异,P-gp表达水平明显下调。两条siR-NA对紫杉醇敏感性的相对逆转率分别达63·8%及51·2%。MDR1基因不同程度的沉默,MDR-A能更有效封闭MDR1基因,MDR1mRNA相对水平下降(67·3±0·8)%。结论siRNA能有效逆转MDR1基因编码P-gp介导的A2780/Taxol细胞多药耐药。Objective To investigate the effect of gene silence of MDR1 gene by small interference RNA （siRNA） in drug resistant A2780/Taxol cell line. Methods From July 2005 to December 2005, two siRNAs which were specifically targeted MDR1 （ Multidrug resistance 1 ） gene were synthesized by Union Hospital of Tongji Medical Schod, then the plasmid was transformed into E. coli. From positive clone, the plasmid was obtained and purified. The purified plasmid was co-transfected with pEGFPN-1 plasmid into .A2780/Taxol cells. The positive expression of transfection was determined by the flow cytometry （FCM）. 50% inhibition concentration （ IC50 ） of paclitaxel on A2780/Taxol was determined by MTr method in vitro. P-gp protein was detected by Western blot. MDR1 mRNA was assessed by RT-PCR. Results The transfection efficiency had no difference between the two constitutive plasmids and the EGFP plasmid. The relative reversal efficiency of A2780/Taxol cells to paclixtel for two siRNA was 63.8 % and 51.2% respectively. The expression of p- gp decreased greatly. Treatment of A2780/Taxol cell with the 2 kinds of siRNAs resulted in a silence of MDR1 gene to different extent. The MDR-A siRNA was more effective in the suppression of MDR1 with a significant reduction of （67.3±0. 8）% of the MDR1 mRNA expression. Conclusion The siRNA can effectively reverse the MDR1/P-gp-mediated multidrug resistance of A2780/Taxol cells.

关 键 词：卵巢肿瘤 小分子干扰RNA MDR1基因 多药耐药 

分 类 号：R71[医药卫生—妇产科学]