【摘要】 目的 研究利用RNA干扰(RNAi)技术来逆转人白血病K562/ADM细胞多药耐药能力的可行性。方法 设计两条针对MDR1基因的发夹状RNA(shRNA)，并将其分别构建在pGenSil-1质粒中；然后将这两条shRNA分别转染K562/ADM。利用罗丹明外排法、荧光定量PCR来检测这两条shRNA对K562/ADM的多药耐药性的逆转作用。结果 在稳定转染的细胞中可以观察到不同程度的MDR逆转现象。在两个稳定转染的克隆中，MDR现象被完全逆转。结论 本研究显示可以利用shRNA逆转K562/ADM细胞的多药耐药问题。

    【关键词】 多药耐药；白血病；shRNA;MDR1

    The complete reversal of multidrug resistance in leukemia cells by shRNA

    ZHANG Xue-qin,CHEN Huan,LI Wen-yi.The Songgang People's Hospital,Bao'an District,Shenzhen 518105,China

    【Abstract】 Objective To study the efficiency of reversing MDR with suppressing MDR1 gene with RNAi in K562/ADM cells.Methods For reversing MDR by RNAi technology,two different short hairpin RNAs (shRNAs) were designed and constructed into pGenSil-1 plasmid respectively.They were then transected into the highly adriamycin-resistant K562/ADM cell line.The RNAi effects on MDR was evaluated by real-time PCR,and Rhodamine123 (Rh123) efflux assay.Results The stable transfected clones showed various degrees of reversal of MDR phenotype.Surprisingly,the MDR phenotype was completely reversed in two transected clones.Conclusions This study demonstrates that MDR can be reversed by the shRNA-mediated RNAi in K562/ADM cells,which provides a valuable clue as to sensitizing multidrug-resistant hepatoma cells to anti-cancer drugs. ......