多药耐药基因修饰的小鼠骨髓细胞对造血功能的保护作用

Protective effect of the bone marrow cells transfected with multidrug resistance gene on the reconstruction of murine hematopoietic function

目的 :将体外转染了人多药耐药基因 (MDR1 )的小鼠骨髓细胞 ,移植给经致死剂量照射的受体小鼠 ,观察该基因对小鼠造血功能的保护作用。方法 :分离经 5 -Fu预处理的供体小鼠骨髓有核细胞 ,体外转染由逆转录病毒介导的人多药耐药基因MDR1 ,然后移植给经 8 5Gy致死剂量照射的同系受体小鼠 ,以紫杉醇 (Taxol)、长春新碱(VCR)、柔红霉素 (DNR)筛选 ,观察小鼠血象、生存期和生存率变化 ,应用聚合酶链反应 (PCR)和流式细胞仪 (FCM)分析人多药耐药基因在小鼠中的整合与表达。结果 :致死剂量辐照后 ,移植组小鼠造血功能逐渐恢复 ,未移植组 1 5d内全部死亡。腹腔注射紫杉醇或静脉注射长春新碱、柔红霉素后 ,实验组生存率和生存期明显高于对照组 (P <0 0 5)。表明MDR1基因能保护骨髓细胞 ,并且具有体内选择和富集作用 ,PCR分析提示 ,实验组外周血、骨髓、肝、脾组织中均检测到原病毒整合 ,RT -PCR与FCM检测到MDR1基因表达。结论 :人MDR1基因修饰的小鼠骨髓细胞 ,能有效重建经致死剂量照射的受体小鼠造血功能 。

AIM: To investigate the protective effect of the bone marrow cells transfected with human multidrug resistance gene (MDR1) on the reconstruction of murine hematopoietic function.METHODS: The mononuclear cells of the bone marrow from donors, BALB/C mice, treated with 5-Fu previously, were isolated and transfected with human multidrug resistance gene in vitro , then transplanted to the tertiary recipients. After lethal irradiation(8.5 Gy) and bone marrow transplantation, the recipients were selected with Taxol 7 mg/kg intraperitoneal injection, VCR 5 mg/kg or DNA 5 mg/kg intravenous injection. The survival rate and blood pictures of mice as well as the integration and expression of target gene MDR1 were studied. RESULTS: The lethal irradiated murine hematopoietic function could be reconstructed and protected from toxicity of high doses Taxol, VCR and DNR selection after reinfusing the hematopoietic progenitor cells containing human multidrug resistance gene (MDR1). The survival rate and survival time of experimental mice were higher than that in the control group. The integration and expression of MDR1 gene in recipients were confirmed by PCR, RT-PCR and FCM. CONCLUSION: The integration and expression of human multidrug resistance gene in recipients may play an important role in the reconstruction and protection of murine hematopoietic function.