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目的:将人胞苷脱氨基酶基因(CD)导入小鼠骨髓细胞后,观察小鼠对大剂量阿糖胞苷(Ara—C)的耐受性,探讨骨髓耐受联合化疗的可行性. 方法:以反转录病毒为载体,将人胞苷脱氨基酶基因(CD)通过共培养转染入小鼠骨髓干细胞,观察共培养后的骨髓细胞及受体小鼠骨髓移植后经药物处理后的骨髓细胞耐Ara—C CFU—GM生成情况;从转基因小鼠骨髓细胞提取DNA,用PCR检测转基因小鼠骨髓细胞耐药基因的表达;观察转基因小鼠经大剂量Ara—C化疗后血象、体质量及生存率的变化. 结果:骨髓移植前共培养后供体的骨髓细胞和骨髓移植后受体含有耐药基因(SFG—CD)的骨髓细胞均有耐药克隆的形成(52%,54%;与对照组相比X2分别为124.62, 126.26,P均<0.01),并明显增加了对Ara—C的耐受性:与对照组比较含耐药基因组动物经大剂量化疗后, 生存率明显提高(X2=7.42,P<0.01),血象、体质量下降幅度较小而恢复较快;转基因小鼠骨髓细胞经PCR检测,显示有CD基因条带. 结论:耐药基因可以进入小鼠骨髓细胞并且获得共表达,提高了造血细胞对Aar—C的耐受性.
OBJECTIVE: To observe the tolerance of mice to high-dose cytarabine (Ara-C) and to investigate the feasibility of combined chemotherapy of bone marrow tolerance after the introduction of human cytidine deaminase gene (CD) into mouse bone marrow cells. METHODS: Recombinant human cytidine deaminase gene (CD) was transfected into mouse bone marrow stem cells by co-culture using retrovirus as a vector. The co-cultured bone marrow cells and recipient mice were treated with bone marrow after drug treatment The bone marrow cells were resistant to Ara-C CFU-GM production. DNA was extracted from the bone marrow cells of transgenic mice and the expression of multidrug resistance genes in bone marrow cells of transgenic mice was detected by PCR. , Body weight and survival rate.Results: The bone marrow cells from donor co-cultured before bone marrow transplantation and the bone marrow cells from recipients with bone marrow resistance after transplantation (SFG-CD) all had resistant clone formation (52% , 54% compared with the control group; X2 was 124.62, 126.26 respectively, P <0.01), and significantly increased the tolerance to Ara-C: Compared with the control group, Survival rate was significantly increased (X2 = 7.42, P <0.01), blood, body weight decreased less Faster recovery; transgenic mouse bone marrow cells by PCR, showed CD gene band Conclusion: resistance gene into the bone marrow cells can be obtained and co-expressed, the hematopoietic cells to improve resistance to the Aar-C.