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细胞色素氧化酶P450 3A4基因联合环磷酰胺的体外抗肿瘤效应
引用本文:张巍,王季石,杨远,方琴. 细胞色素氧化酶P450 3A4基因联合环磷酰胺的体外抗肿瘤效应[J]. 复旦学报(医学版), 2009, 36(4): 384-388. DOI:  
作者姓名:张巍  王季石  杨远  方琴
作者单位:1. 贵阳医学院第一附属医院血液科,贵阳,5501104
2. 贵阳医学院第一附属医院药剂科,贵阳,5501104
基金项目:国家自然科学基金项目 
摘    要:目的 克隆细胞色素氧化酶P450 3A4(CYP3A4)基因,构建真核表达质粒并导入K562细胞进行表达,研究CYP3A4联合环磷酰胺(CPA)的体外抗肿瘤效应。方法 利用RT-PCR从人肝细胞中克隆CYP3A4基因,构建重组真核表达载体。脂质体法导入K562细胞,RT-PCR和Western blot检测CYP3A4基因在K562细胞中的表达并筛选稳定转染的细胞克隆。MTT法检测CYP3A4联合CPA对肿瘤细胞的抑制作用。结果 成功克隆CYP3A4基因并构建了真核表达质粒pcDNA3.1/myc-His A(+)-CYP3A4。RT-PCR和Western blot分别显示转基因组的CYP3A4 mRNA和蛋白质表达水平均显著高于转空载体组和对照组。MTT示转基因组IC50值为1.090 16 mmol/L,明显低于转空载体组和对照组。酶诱导剂和抑制剂分别能够减低和增高转基因组IC50。结论 CYP3A4体外具有联合CPA的抗肿瘤作用,这种作用能够被CYP3A4相应的诱导剂和抑制剂增强或减低。

关 键 词:细胞色素氧化酶P450 3A4;环磷酰胺;基因介导的酶前药治疗;肿瘤治疗;基因转染
收稿时间:2009-01-16

The anti-tumor effect of Cytochrome oxidase P450 3A4 combined with Cyclophosphamide in vitro
ZHANG Wei,WANG Ji-shi,YANG Yuan,FANG Qin. The anti-tumor effect of Cytochrome oxidase P450 3A4 combined with Cyclophosphamide in vitro[J]. Fudan University Journal of Medical Sciences, 2009, 36(4): 384-388. DOI:  
Authors:ZHANG Wei  WANG Ji-shi  YANG Yuan  FANG Qin
Abstract:Objective To clone CYP3A4 gene and to construct CYP3A4 recombinant mammalian expression vectors which are transferred into K562 cells for expression,and detect the anti-tumor effect of CYP3A4 combined with Cyclophosphamide (CPA) in vitro.Methods Mammalian expression vector containing CYP3A4 gene cloned from human hepatocytes by RT-PCR were constructed and transferred into K562 cells via liposome. The expression of CYP3A4 was detected by RT-PCR and Western blot respectively. MTT detected the anti-tumor effect of CYP3A4 recombinant mammalian expression vectors combined with CPA.Results We cloned CYP3A4 gene and constructed the recombinant mammalian expression vectors pcDNA3, 1/myc-His A (+)-CYP3A4 successfully. Both RT-PCR and Western blot showed significantly higher mRNA and protein expression of CYP3A4 in gene-transfected group than in empty vector-transfected controls and in empty cells controls. The IC_(50) values were 1. 090 16 mmol/L in gene-transfected group, which were significantly lower than in other two groups and could be reduced and increased by the revulsant and inhibitor respectively.Conclusions CYP3A4 recombinant mammalian expression vectors had anti-tumor effect cooperating with CPA, and the effect could be increased and reduced by the revulsant and inhibitor of CYP3A4 respectively.
Keywords:CYP3A4  cyclophosphamide  gene directed enzyme prodrug therapy  neoplasm therapy  gene transfection
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