SiRNA逆转肺耐药相关蛋白表达介导的白血病细胞多药耐药

Reversal of muti-drug resistance in leukemia cells induced by lung resistance-related protein by short interfering RNA

目的:研究双链短干扰RNA(siRNA)对白血病多药耐药细胞模型(K562/NaB)肺耐药相关蛋白(LRP)表达及功能的影响.方法:针对LRP基因设计合成特异性siRNA,在脂质体介导下转染K562/NaB;采用半定量逆转录聚合酶链反应(RT-PCR)检测K562细胞LRP mRNA的水平;用流式细胞术检测K562/NaB细胞LRP蛋白表达的变化和细胞内柔红霉素(DNR)的蓄积;MTT法检测阿霉素(ADM)对K562/NaB细胞耐药的半数抑制浓度(IC50).结果:siRNA转染后:K562/NaB细胞的LRPmRMA水平明显降低;LRP蛋白表达由阳性转为阴性;细胞内DNR的蓄积明显增加,DNR平均荧光增强3.28倍;对ADM药物敏感相对逆转效率为78.18%.结论:siRNA可逆转由LRP介导的白血病细胞多药耐药.

AIM： To investigate the effect of short interfering RNA （siRNA） on expression and function of lung resistancerelated protein （LRP） in the multi-drug resistant human leukemia cells （K562/NaB）. METHODS： Multidrug-resistant K562 cells with high level LRP expression treated with sodium butyrate （NaB）, was used as an in vitro model system. LRP specific siRNA was synthesized and transfected into the K562/NaB cells. Expression of LRP mRNA was detected by reverse transcriptasepolymerase chain reaction （ RT-PCR）, and protein level and intracellular daunorubicin （DNR） accumulation in K562/NaB cells were detected by flow cytometry. 50% inhibition concentration （IC50） of adriamycin （ ADM ） on K562/NaB cells was detected by MTY method. RESULTS： LRP mRNA level was decreased obviously; the protein expression was turned from positive result to negative result. Intracellular DNR accumulation was increased and the mean fluorescence of DNR was 3.28 times higher. The relative efficiency to ADM was 78.18%. CONCLUSION： The siRNA could effectively reverse the multi-drug resistance of leukemia cells induced by LRP.