人多药耐药基因-1及二氢叶酸还原酶基因共转导小鼠造血细胞  

Cotransduction of Human mdr-1 and Dihydrofolate Reductase Genes into Murine Hematopoietic Cells

作　　者：刘晓丹 于晓妉[1] 郭子宽[1] 李秀森[1] 刘元林[1] 毛宁[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《中国实验血液学杂志》2000年第2期110-113,共4页Journal of Experimental Hematology

摘　　要：耐药基因转导骨髓细胞对化疗病人造血系统的保护作用已受到人们的重视。本研究利用多药耐药基因 1(mdr 1)及二氢叶酸还原酶 (dhfr)双基因转染小鼠骨髓细胞 ,观察造血细胞对两种耐药谱化疗药物的抵抗能力。结果表明 ,所用逆转录病毒载体转染率达到 15 %左右 ,转基因骨髓细胞CFU GM对taxol及MTX的耐药能力明显增强 ,说明外源基因在造血祖细胞中的正确表达 ;转基因骨髓细胞回输给同系小鼠 7个月后 ,FCM技术仍可检测到外周血白细胞 gp170的表达 ,且其基因组DNA中可检测到mdr 1及dhfr基因特异序列 ,表明两种基因可能已稳定整合至造血干 /祖细胞中。上述结果为以骨髓细胞为靶体的耐药基因治疗提供了有用的实验室资料。Transfer of drug resistance genes into hematopoietic cells is an attractive approach to protect hematopoietic system from the toxic effects by chemotherapeutic agents in cancer patients.In this study,transduction of mdr 1 in combination with dihydrofolate reductase(dhfr) gene was performed, and the expression of exogenous genes and chemoprotection capacity in mouse bone marrow cells were observed. The results showed that approximately 15% of bone marrow cells transfected with the retroviral vector expressed mdr 1 as assayed by flow cytometry. Gene transfer resulted in about 0.9-13 fold and 0.5-2.6 fold increase in the resistance of CFU GM to taxol and methotrexate in vitro, respectively (P<0.05). Moreover, seven months after transplantation to syngeneic mice with mdr 1 and dhfr transfected bone marrow cells, peripheral blood cells in recipients were still positive for gp170 as evaluated by FACS as well as for mdr 1 and dhfr by PCR amplification. These results indicate that hematopoietic progenitors can be transfected by retrovirus containing mdr 1 and dhfr genes, and that functional drug resistance accompanies their expressions. Furthermore, genetic chimerism might exist in hematopoietic stem cells. In conclusion, transfer and expression of mdr-1 and dhfr genes in bone marrow cells might be applicable in gene therapy research in cancer patients.

关 键 词：耐药基因 二氢叶酸还原酶基因 造血细胞 肿瘤 

分 类 号：R73-362[医药卫生—肿瘤]