| 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 |
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| 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: |
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| 作者姓名： | 张巍 王季石 杨远 方琴 |
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| 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 |
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| 基金项目： | 国家自然科学基金项目 |
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| 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。
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| 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 |
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| 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: |
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| Authors: | ZHANG Wei WANG Ji-shi YANG Yuan FANG Qin |
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| 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. |
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| Keywords: | CYP3A4 cyclophosphamide gene directed enzyme prodrug therapy neoplasm therapy gene transfection |
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