【摘要】 目的检测转染多药耐药基因-1(multidrug resistance gene-1,mdr-1)的骨髓细胞能否耐受大剂量霉酚酸脂(mycophenolate mofetil,MMF)的杀伤作用,为体内实验提供依据。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;MTT法检测不同浓度MMF处理后转染mdr-1基因骨髓细胞的存活情况。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-1表达产物P-gp有药物外排泵功能;不同剂量的MMF处理后,转染mdr-1基因的骨髓细胞存活率没有明显差异(P>0.05)。结论转染mdr-1的大鼠骨髓细胞能有效耐受超大剂量MMF治疗。更多还原

【Abstract】 Objective To investigate whether bone marrow cells(BMC) transfected with multidrug resistance gene-1(mdr-1) can tolerate to the killing effect of overdose mycophenolate mofetil(MMF).Methods BMC were transfected with recombinant adenovirus,Ad-EGFP-mdr-1.Expression levels of mdr1 mRNA and protein were detected by RT-PCR,Western blotting and daunorubicin(DAM),respectively.The survival of mdr-1 transfected BMC after treatment with MMF at different concentrations was detected by MTT.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.No obvious difference was found in survival of mdr-1 transfected BMC(P>0.05).Conclusion mdr-1 transfected MBC can efficiently tolerate to overdose MMF,thus providing a basis of its in vivo experiments.更多还原