The anti-tumor effect of Cytochrome oxidase P450 3A4 combined with Cyclophosphamide in vitro
- VernacularTitle: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
- Author:
Wei ZHANG
;
Jishi WANG
;
Yuan YANG
;
Qin FANG
- Publication Type:Journal Article
- Keywords:
CYP3A4;
cyclophosphamide;
gene directed enzyme prodrug therapy;
neoplasm therapy;
gene transfection
- From:
Fudan University Journal of Medical Sciences
2009;36(4):384-388
- CountryChina
- Language:Chinese
-
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.
