mdr1单因素耐药白血病细胞株的构建

目的 构建mdrl单因素耐药白血病细胞株，为研究RNA干扰逆转mdr1所致耐药提供细胞模型。方法 将含mdr1 cDNA全长的真核表达载体通过脂质体转染人对化疗药物敏感的K562细胞内，G418筛选获得转基因单克隆细胞．用RT-PCR检测有无mdr1表达、流式细胞技术检测细胞表面P-gp蛋白含量、Rh123泵出实验检测P-gp功能、MTT检测细胞对药物的敏感性。结果 转mdr1基因细胞K562／mdr1能较高水平表达mdr1，明显表现出对化疗药物的耐药性。结论 通过转基因方法构建mdr1单因素耐药细胞株可为RNA干扰逆转mdr1提供更精确的细胞研究模型。

Establishment of a Drug Resistant Leukemia Cell Line Caused by mdr1 Gene

Objective To establish a drug resistant leukemia cell line caused by mdr1 gene for the mdr1 reversing research. Methods The leukemia sensitive cell line K562 was transfected with mdr1 cDNA by liposome-induced gene transfection and selected by G418. The mdr1 phenotype was identified by RT-PCR, the P-gp expression was detected by flow cytometry, the function of P-gp was measured by Rh123 efflux experiment, and the sensitivity of cell lines to the drug was detected by MTT test. Results The transfected K562/mdr1 with mdr1 cDNA highly expressed mdr1 and showed a remarkable drug resistant feature. Conclusion mdr1 plays an important role in drug resistance and the mdr1 cell line established by gene transfection can provide a better cell model for mdr1 reversing research.