重组腺病毒Ad-EGFP-mdr-1转染大鼠骨髓细胞模型的构建

Construction of recombinant adenovirus Ad-EGFP-mdr-1 transfected bone marrow cells

目的构建转染多药耐药基因-1(multidrug resistance gene-1,mdr-1)的骨髓细胞。方法将重组腺病毒Ad-EGFP-mdr-1转染大鼠骨髓细胞。RT-PCR法和Western blot法检测mdr-1 mRNA及蛋白的表达水平;柔红霉素排出试验检测转染mdr-1基因后的骨髓细胞有无功能表达。结果骨髓细胞病毒转染率为30.14%,存活率为89.25%,转染后骨髓细胞mdr-1的mRNA及蛋白质水平都存在功能性表达,导入的mdr-I表达产物P-gp有药物外排泵功能。结论转染mdr-1的大鼠骨髓细胞模型的成功构建,为进一步研究肿瘤的多药耐药提供实验基础。

Objective To construct bone marrow cells （BMC） transfected with adenovirus Ad - EGFP - mdr - 1 （ multidrug resistance gene - 1 ）. Methods BMC were transfected with recombinant adenovirus, Ad - EGFP - mdr - 1. Ex- pression levels of mdr - 1 mRNA and protein were detected by RT - PCR, Western blotting and daunorubicin （DAM）. Results The transfection efficiency of the virus was 30. 14% ,and the survival rate of BMC was 89.25%. The mdr - 1 mRNA and protein were functionally expressed in transfected BMC. The expressed product, P - gp, was found to have a pumping function induced by mdr - 1. Conclusion Constructing the bone marrow cells were transfected recombinant adenovirus Ad - EGFP - mdr - 1 helps to further study reversing multidrug resistance.