摘要
目的探讨人双突变的二氢叶酸还原酶(DHFR)基因对小鼠化疗保护作用。方法以反转录病毒为载体,将DHFR基因转染入小鼠骨髓干细胞,观察氨甲喋呤(MTX)处理后的骨髓细胞中粒细胞-巨噬细胞克隆形成单位(CFU-GM)的生成情况;观察大剂量MTX化疗后转基因小鼠血象、体重及生存率的变化;用RT-PCR检测转基因小鼠骨髓细胞耐药基因的表达。结果转染SFG-F／S- NeoR耐药基因的骨髓细胞有耐药克隆的形成,供体小鼠为15．8％,受体小鼠为18．0％,对照组为0;大剂量化疗后,含耐药基因组小鼠血象、体重逐渐恢复正常,生存率为83．3％(第40天),对照组为0;转基因小鼠骨髓细胞经RT-PCR检测,显示有F／S基因条带(400 bp)。结论DHFR耐药基因可导入小鼠骨髓细胞并获得表达,提高了骨髓细胞对MTX的耐药性。
Objective To explore the feasibility of transfecting DHFR （ human double-mutant dihydrofolate reductase） gene into mouse bone marrow cells and the effect of resistance to high dose MTX chemotherapy. Methods After DHFR gene was transfected into mouse bone marrow cells with retroviral vector, the cells were treated with methotrexate （MTX） and then CFU-GM （ granulocyte-macrophage colonyforming unit） assay was performed. Peripheral blood leucocytes and platelets, body weight and survival rate were observed. After treatment with high dose MTX, the expression of drug resistance gene was checked by RT-PCR in the transfected bone marrow cells. Results SFG-F/S-NeoR gene-transfected mice bone marrow cells yielded drug-resistance colonies to MTX （donor mice： 15.8% , recipient mice： 18.0% , control： 0）. The peripheral blood leucocytes and platelets, body weight recovered gradually and the survival rate was 83.3% at the 40th day, while 0 in controls in gene transfected mice after large dose MTX treatment. RTPCR of transgenic mouse marrow cells showed the band of F/S gene （400 bp）. Conclusion DHFR gene can not only be integrated and expressed in bone marrow cells but also improve their drug-resistence to MTX.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2006年第8期583-585,共3页
Chinese Journal of Oncology
作者简介
通讯作者：路平，E-mail：lupingllll@yahoo．com
