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目的探讨MDR1基因转染荷瘤小鼠骨髓单个核细胞(BM-MNCs)后在体内的分布和对骨髓造血细胞的作用。方法将32只H22肝癌荷瘤BALB/c小鼠随机分为4组:正常对照组(不照射不回输)、空白对照组(照射并回输生理盐水)、阴性对照组(照射并回输未转染的BM-MNCs)、转染实验组(照射并回输已转染的BM-MNCs),每组8只。阿霉素超剂量化疗,监测化疗过程中各组外周血细胞和肿瘤体积的变化,用RT-PCR法和免疫组化法检测外源性人MDR1基因和P-gp在荷瘤小鼠体内的表达和分布。结果化疗后各组红细胞和血小板均无明显变化,差异无统计学意义(P>0.05);转染实验组白细胞数于化疗后第4周降至(3.32±0.26),与化疗前(6.64±0.39)相比减少有统计学意义(P<0.05),化疗后第3周起转染实验组白细胞数高于阴性对照组,增高有统计学意义(P<0.05);化疗3周后与3个对照组相比转染实验组肿瘤体积较小,减小有统计学意义(P<0.05);外源性人MDR1基因和P-gp在骨髓和外周血单个核细胞有表达和分布,在心、肝、肺、脾和肾等组织中无表达和分布;肿瘤组织中无外源性人MDR1基因分布,有P-gp表达。结论MDR1基因转染荷瘤小鼠BM-MNCs后在超剂量化疗中对骨髓造血功能有保护作用,外源性MDR1基因在心、肝、肺、脾和肾等组织中无表达。
Objective To investigate the distribution of MDR1 gene in bone marrow-derived bone marrow-derived mononuclear cells (BM-MNCs) in vivo and its effect on bone marrow hematopoietic cells. Methods Twenty-two H22 hepatocellular carcinoma-bearing BALB / c mice were randomly divided into 4 groups: normal control group (no irradiation without return), blank control group (irradiation and saline return), negative control group Untransfected BM-MNCs) were transfected into the experimental group (irradiated and transfused transplanted BM-MNCs), 8 in each group. Adriamycin overdose chemotherapy to monitor the changes of peripheral blood cells and tumor volume in each group during chemotherapy, and the expression of exogenous human MDR1 gene and P-gp in tumor-bearing mice was detected by RT-PCR and immunohistochemistry And distribution. Results After chemotherapy, there was no significant difference in erythrocytes and platelets between the two groups (P> 0.05). The number of leukocytes in transfection group decreased to (3.32 ± 0.26) in the fourth week after chemotherapy, 0.39) (P <0.05). The number of leukocytes in the transfection experiment group from the third week after chemotherapy was higher than that of the negative control group (P <0.05). After 3 weeks of chemotherapy, Compared with the transfected group, the tumor volume of the control group was smaller and decreased statistically (P <0.05). Exogenous human MDR1 gene and P-gp were expressed and distributed in the bone marrow and peripheral blood mononuclear cells, , No expression and distribution in liver, lung, spleen, kidney and other tissues. There was no exogenous human MDR1 gene distribution in tumor tissues and P-gp expression. Conclusion The MDR1 gene has the protective effect on the hematopoietic function of bone marrow after transfection with BM-MNCs in tumor-bearing mice. The exogenous MDR1 gene has no expression in heart, liver, lung, spleen and kidney.