MDR1基因修饰的自体骨髓移植后体内表达及时限的研究

目的 研究多药耐药基因1(multidrug resistanle gene 1, MDR1)转染的骨髓单个核细胞自体移植后,外源性MDR1基因在骨髓中的功能性表达及时限.方法 体外浓缩病毒上清转染法将MDR1基因导入兔骨髓单个核细胞;经大剂量环磷酰胺化疗预处理后,将转染的骨髓单个核细胞行自体骨髓移植;采用PCR法、免疫组织化学法和柔红霉素(daunorubicin, DNR)排出试验检测MDR1基因在受体骨髓中的整合及功能性表达.结果 自体骨髓移植后1～4个月,PCR法检测到外源性MDR1基因在骨髓细胞基因组中的整合;免疫组化法测得骨髓单个核细胞中P-gp阳性率分别为9.5%、8.5%、6.0%、3.5%;柔红霉素排除试验检测到定植的MDR1基因能功能性的表达.结论 转染MDR1基因的骨髓单个核细胞自体移植后,MDR1基因能定植于骨髓中并功能性的表达4个月.

Functional expression and temporality of MDR1 gene in bone marrow of rabbits after autologous transplantation with the gene modification to mononuclear cells

Objective To explore the functional expression and temporality of MDR1 gene in bone marrow of rabbits after autologous bone marrow transplantation with MDR1 transferred bone marrow mononuclear cells. Methods The supernatant of the amphotropic virus producer cell line PA317-HaMDR1/A was collected and concentrated to cocultivate with the bone marrow mononuclear cells of the rabbits. After large dose of chemotherapy with cyclophosphamide,the transferred cells were autotransplanted into the bone marrow. The integration,transfection rate and physiological function of MDR1 gene were tested by PCR,SP immunohistochemical method and daunorubicin (DNR) extrusion test respectively. Results After autologous bone marrow transplantation had been executed for 1-4 months,the integration of MDR1 gene in genome of bone marrow mononuclear cells was detected by PCR,and the expression rates of P-gp in cells tested by SP immunohistochemical method were 9.5%,8.5%,6.0% and 3.5% respectively. The physiological function of MDR1 gene in bone marrow cells was proved by DNR extrusion test. Conclusion After the autotransplantation with bone marrow mononuclear cells transferred by MDR1 gene,the MDR1 gene can implant into the bone marrow of rabbits and has expressed functionally for 4 months,which has provided a basis for further research on chemoprotection experiment of the MDR1 gene transferred into the bone marrow cells.