论文部分内容阅读
目的研究外源性人多药耐药基因(human multidrug resistance gene 1,hMDR1)转染骨髓单个核细胞(mono-nuclear cells,MNCs)移植联合超剂量表阿霉素化疗在4T1乳腺癌治疗中的骨髓保护和治疗作用。方法用携带目的基因hMDR1和绿色荧光蛋白(green fluorescence protein,GFP)报告基因的复制缺陷型重组腺病毒(rAd-hMDR1-GFP)转染原代培养的MNCs以获取hMDR1基因修饰的MNCs(MNCs-rAd-hMDR1-GFP),将修饰后的MNCs移植到4T1乳腺癌模型体内,在表阿霉素超剂量化疗中观察骨髓造血功能的保护和肿瘤的治疗效果。结果 hMDR1基因成功转入小鼠骨髓单个核细胞,体外转染率达26.26%;转染细胞移植后能定植于受体骨髓中并功能性表达。在超剂量化疗中,hMDR1基因能有效保护受体骨髓的造血功能,荷瘤动物能耐受3倍剂量表阿霉素化疗,外周血白细胞数能维持在(5.7±0.6)×109/L;同时超剂量化疗能迅速杀灭肿瘤细胞,使荷瘤动物存活时间明显延长(P<0.05),治愈率明显提高(P<0.05)。结论 hMDR1基因转染骨髓造血细胞移植能明显提高受体骨髓对化疗药物的耐受性,联合超剂量化疗能快速、有效的杀灭肿瘤细胞,提高恶性肿瘤的治愈率。
Objective To investigate the effect of exogenous human multidrug resistance gene 1 (hMDR1) transfection on bone marrow mononuclear cells (MNCs) transplanted in combination with super-dose epirubicin chemotherapy in the treatment of 4T1 breast cancer Bone marrow protection and therapeutic effects. Methods Primary cultured MNCs were transfected with replication-defective recombinant adenovirus (rAd-hMDR1-GFP) carrying target gene hMDR1 and green fluorescence protein (GFP) reporter gene to obtain hMDR1 gene-modified MNCs (MNCs- rAd-hMDR1-GFP). The modified MNCs were transplanted into the 4T1 breast cancer model. The hematopoietic function of the bone marrow and the therapeutic effect of the tumor were observed in epirubicin overdose chemotherapy. Results The hMDR1 gene was successfully transfected into mouse bone marrow mononuclear cells and the transfection rate was 26.26% in vitro. Transfected cells could be implanted into the bone marrow of recipients and expressed functionally. In overdose chemotherapy, hMDR1 gene can effectively protect the hematopoietic function of the recipient bone marrow. The tumor-bearing animals can tolerate 3 times the dose of epirubicin chemotherapy and the number of peripheral white blood cells can be maintained at (5.7 ± 0.6) × 109 / L. At the same time, overdose chemotherapy can kill tumor cells rapidly, prolong the survival time of tumor-bearing animals significantly (P <0.05), and cure rate was significantly increased (P <0.05). CONCLUSION: Transplantation of hMDR1 gene into bone marrow cells can significantly increase the tolerance of the recipient bone marrow to chemotherapeutic agents. Combining with super-dose chemotherapy, it can kill tumor cells rapidly and effectively and improve the cure rate of malignant tumors.