多药耐药基因(MDR1)转染小鼠骨髓造血细胞表达及功能研究

目的：探索含人多药耐药基因（ＭＤＲ１）全长ｃＤＮＡ的逆转录病毒载体转染小鼠骨髓造血细胞转染时间及转染效率的关系。方法：采用逆转录病毒上清法转染小鼠骨髓造血细胞，依据转染时间不同分为２日组及４日组，分别行ＰＣＲ法检测外源ＭＤＲ１基因；免疫组化法检测ＭＤＲ１基因表达，柔红霉素排出试验检测表达外源基因的功能。结果：ＰＣＲ方法证实转入的外源ＭＤＲ１基因可整合入小鼠骨髓造血细胞基因组中；免疫组化法检测转染率最高达３３％，转染４日组转染率大于转染２日组转染率（Ｐ＜０．０５）；柔红霉素排出试验证实转入外源基因表达产物可行使正常功能。结论：逆转录病毒上清法可有效转染外源ＭＤＲ１基因入骨髓造血细胞中；表达产物可行使正常生理功能；在转染４天时间内，转染率随转染时间延长而增高。

Research on the expression and function of MDR1 gene transferring into the murine hematopoietic cells of bone marrow

Objective: Using retrovirus vector to transfer multidrug resistance gene(MDR1gene) into hematopoietic cells of murine bone marrow, and discussing the relation between transduction rate and the time. Methods: Supernatant method is used to transfer MDR1 gene into hematopoietic cells of murine bone marrow. PCR method is used to detect the gene integration. Immunohistochemistry(IC) method is used to measure the transduction rate. DNR extrusion test is used to measure the function of gene product. Results: Gene integration was tested by PCR method.The transduction rate is as high as 33%,and the rate of 4-day group is higher than that of 2-day group.The product of transferred gene has its physical function. Conclusion: Supernatant method is an effective way to transfer MDR1 gene into hematopoietic cells of murine bone marrow, and the product of MDR1 gene has its phsiological function. The transduction rate elevated following the prolonging of the transduction time during 4 days of transduction.