糖皮质激素受体在白血病CYP3A5耐药中的信号转导作用

Signal transduction effect of glucocorticoid receptor in drug resistance of CYP3A5 in leukemia

目的探讨糖皮质激素受体在白血病CYP3A5耐药中的信号转导作用。方法采用DNA重组技术构建ptracer-hGR重组质粒,并用报告基因检测方法研究糖皮质激素受体对CYP3A5药物代谢酶转录、表达调控中的信号转导作用。结果菌落PCR及测序证实成功构建ptracer-hGR重组质粒。经阿糖胞苷、甲氨蝶呤和长春新碱三种化疗药物加药后共转染ptracer-hGR质粒的实验组荧光素酶相对活性显著高于单纯转染CYP3A5报告基因质粒的对照组,差异具有统计学意义(P=0.0362、0.0034和0.0164);而卡铂和米托蒽醌加药后,荧光素酶相对活性无明显增高趋势。结论阿糖胞苷、甲氨蝶呤和长春新碱三种化疗药物可能通过激活糖皮质激素受体这一信号途径而上调CYP3A5 mRNA的转录。

Objective To investigate the signal transduction effect of glucocorticoid receptor in the drug resistance of CYP3A5 in leukemia. Methods The ptracer-hGR recombinant plasmid was constructed using DNA recombinant technology,and the signal transduction effect of glucocorticoid receptor in the regulation of drugs on the transcription and expression of CYP3A5 was analysed by reporter gene assay. Results Colony PCR and sequencing indicated that the ptracer-hGR recombinant plasmid was constructed successfully.The relative luciferase activity in the test cells co-transfected with ptracer-hGR was remarkably higher than that in the control cells transfected with pGL3-CYP3A5 promotor alone after the cells were subjected to the chemotherapy drugs such as cytosine arabinoside（Ara-C）,methotrexate（MTX） and vincristine（VCR）（P=0.036 2,0.003 4 and 0.016 4,respectively）.However,the relative luciferase activity had either no or only a slight increase after the cells were subjected to the other chemotherapy drugs such as carboplatin and mitoxantrone. ConclusionAra-C,MTX and VCR may up-regulate the transcription of CYP3A5 mRNA by activation of glucocorticoid receptor.