mdr-1基因转染的骨髓细胞对大剂量MMF治疗的耐受性研究  

Tolerance of mdr-1 transfected bone marrow cells to overdose mycophenolate mofetil

作　　者：李钺[1] 栾晓峰[1] 陈勇[1] 龚建平[1] 陈先锋[1] 丁雄[1] 刘作金[1] 

机构地区：[1]重庆医科大学附属第二医院肝胆外科,重庆市肝胆外科重点实验室,重庆400010

出　　处：《第三军医大学学报》2010年第3期242-245,共4页Journal of Third Military Medical University

基　　金：中国博士后科学基金(20070411151);重庆市自然科学基金(CSTC2007BB0266;CSTC2008BB5229);重庆市涪陵区自然科学基金(2008-48)~~

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。Objective To investigate whether bone marrow cells （BMC） transfected with muhidrug 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 mdrl 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）. Gonclusion mdr-1 transfected MBC can efficiently tolerate to overdose MMF, thus providing a basis of its in vivo experiments.

关 键 词：多药耐药基因-1 霉酚酸脂 骨髓细胞 

分 类 号：R331.22[医药卫生—人体生理学] R394.2[医药卫生—基础医学]