逆转录病毒载体介导MDR1基因转染人外周血造血干细胞的研究

目的：多药耐药基因（ＭＤＲ１）的异常扩增和过度表达是肿瘤细胞产生多药抗药性的重要原因之一。本研究试图将ＭＤＲ１导入骨髓或外周血造知干细胞并使入获得耐药表型，以用于减轻大剂量化疗对造血细胞的毒性。方法：应用反转录病毒载体ｐＨａＭＤＲ１／Ａ将ＭＤＲ１阳性。ＰＣＲ法检测转染细胞所 ＣＦＵ－ＧＭ落集中，外源ＭＤＲ１阳性比例为１／４；而未转染细胞所形成的ＣＦＵ－ＧＭ集匀为阴性。集落形成试验证明，转染和非转染

Retroviral Vector - Mediated Introduction of MDR1 cDNA Into Human Hematopoietic Stem Cells form Peripheral Blood

Objective: Over-experession of human multi-drug resistance gene ( MDR1) is one of major mechanisms for resistance of tumor cells to many naturally-occuring anticancer agents. In this study, authors tried to get introduction of exogenous MDR1 into hematopoietic stem celk which might confer higher level of expression of P170 in these cells and alleviate the marrow toxicity of high dose chemotherapy. Method: By using retrovial vector (pHa MDR1/A) to introduce MDR1 cDNA into human hematopoietic stemcells. Results:The authors successfully transfered MDR1 cDNA into mobilized hematopoietic progenitors form the peripheral blood of a patient with breast cancer using the retrovial vecto. The overall transduction efficiency was about 3.74%, measured by flowcytometry. PCR assays proved that 1/4 of CFU - GM formed by transduced stem cells were exogenous MDR1-positive. Colony formation assays also confirmd that MDR1 - transduced hematopoietic progenitors were more resistant to treatment of Taxol compared with the untransduced cells. Conclusion: These preliminary results suggest the feasibility of retroviral vector-introduced MDR1 cDNA into human hematopoietic stem cells.