目的：观察腺相关病毒载体介导外源基因导入人外周血干细胞的转染效率及表达。方法：本文采用腺相关病毒(adeno-associated virus, AAV)载体介导的基因转移方法，将绿色荧光蛋白（green fluorescent protein, GFP）基因和人多药耐药基因（multi-drug resitance, MDR1）导入人外周造血干/祖细胞，并对转染效率和转基因造血细胞的耐药性进行了初步研究。结果：rAAV/GFP感染CD34阳性细胞后，在荧光显微镜下观察，约30%细胞中可见绿色荧光。将rAAV/MDR1重组病毒感染CD34阳性细胞，经PCR和MTT法证实，导入MDR1基因的CD34阳性细胞对秋水仙素的耐药性与未导入MDR1基因的细胞有明显差异。结论： 腺相关病毒载体能有效地将外源基因导入外周血CD34阳性细胞，并可在其中有效地表达。

Objective: To investigate the expression of MDR1 and GFP in the human hematopoietic cells mediated by adeno-associated virus. Methods:The GFP gene was transferred into the human hematopoietic cells by AAV vectors and created strong visible fluorescence by purely molecular biological means. Using adeno-associated virus vectors, we have transferred human mdr-1 gene into human hematopoietic cells and investigated the drug resistence of human hematopoietic cells modified with mdr-1 gene. PCR analysis confirmed that mdr1 cDNA had been successfully transferred into the human hematopoietic cells.An assay of MTT proved that the human hematopoietic cells modified by mdr1 gene had resistance to colchicine.Results: It was about 30% of the hematopoietic cells that expressed the green fluorescent proteins. The resistance of hematopoietic cells was increased parently when the cells were infected by the crude virus stocks. Conclusion: It is conducted that the AAV vector could successfully transfer the foreign gene into the human hematopoietic cells. The cells modified with mdr1 gene have increased the resistance to drugs.

国家自然科学基金（39270826）资助