多药耐药基因转染人骨髓间充质干细胞对其化疗保护作用的研究  

The transfection of multidrug resistance gene(mdr1) into bone marrow mesenchymal stem cells and reinforcing the resistance to chemotherapy

作　　者：吴冰[1] 陈慧玲[1] 王存邦[1] 白海[1] 

机构地区：[1]兰州军区兰州总医院血液病研究所,甘肃兰州730050

出　　处：《现代肿瘤医学》2011年第2期248-251,共4页Journal of Modern Oncology

摘　　要：目的:慢病毒介导的多药耐药基因(mdr1)转染人骨髓间充质干细胞(hBMSCs),探讨其对化疗药物的耐受性。方法:采用percoll密度离心法自骨髓中分离MSCs并进行纯化和扩增,流式细胞仪鉴定。克隆mdr1基因,构建慢病毒载体,命名为TG-005-mdr1/tap。脂质体转染法转染携带mdr1基因的逆转录病毒载体导入293T包装细胞,获得的病毒上清感染hMSCs,GTP荧光技术和western-blot检测mdr1基因的表达,MTT法检测hMSCs对化疗药物的耐受性。结果:成功构建携带mdr1基因的慢病毒载体,采用GTP荧光技术检测慢病毒转染率达83.44%,western-blot检测mdr1编码产物P-pg蛋白高表达,转染后hMSCs对化疗药物的耐受性明显高于对照组。结论:转染mdr1基因后的hMSCs具有化疗耐受性,可为化疗骨髓防护提供新思路。Objective：Multidrug resistance gene （ mdr1 ） transfected into the bone marrow mesenchymal stem cells （hBMSCs） mediated by lentiviral vector, then investigated the resistance of MSCs to chemotherapy. Methods： hBMSCs were isolated and enriched by Percoll density gradient, then identified by flow cytometer. Cloned mdr1 and constructed its ]entiviral vector named TG -005 - mdr1/tap. The lentiviral vector carrying mdr1 was transfected to the packaging cell 293T by lipofeetamine mediated gene transfer. MSCs were transfected by lentiviral vector. GTP marker and western - blot were used to detect the expression of mdr1, and the resistance of MSCs to chemotherapy were detected by MTT assay. Results： Constrcted TG -005 -mdr1/tap successfully, GTP marker detect the efficiency of transfection was 83.44% ;western -blot detect the high expression of P-gp protein. There was a significant increasing in chemoresistant of the transfected MSCs compared with untransfected MSCs. Conclusion： The hMSCs that transfected by mdr1 have higher resistance to chemotherapy. That may provide a new thought to the protection of bone marrow.

关 键 词：多药耐药基因 慢病毒载体 间充质干细胞 转染 化疗耐受 

分 类 号：R73-36[医药卫生—肿瘤]