[Stable expression of human cytochrome P450 2E1 cDNA in nasopharyngeal cancer cell line CNE-2].
OBJECTIVE
This study as designed to investigate the function of CYP2E1 protein in chemical carcinogen metabolism in vitro and its action mechanisms in nasopharyngeal carcinogenesis.


METHODS
DAN recombination technique was used to construct a eukaryotic expression vector pcDNA3.1-2E1 of human cytochrome P450 2E1 cDNA. The vector was introduced into a nasopharyngeal cancer cell line CNE-2 mediating by lipofectamine. After these transforming cells were treated with G418 for 2 weeks, several G418-resistant clones were selected and amplified in culture.


RESULTS
The authors acquired two cell clones CNE2-2E1-1 and CNE2-2E1-2 with stable integration and expression of exogenous CYPE2E1 cDNA. After Southern blot analysis, RT-PCR and Western blot were fulfilled.


CONCLUSIONS
The cell models setting here will be a good tool in the further study of CYP2E1. At the same time, its use will help to screen chemical carcinogenesis and to elucidate the mechanism of carcinogenesis which is related to chemical carcinogen activated by CYP2E1.
Wang
S L
SL
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, China.
He
Z M
ZM
Chen
Z C
ZC
D002273:Carcinogens; D003001:Cloning, Molecular; D019392:Cytochrome P-450 CYP2E1 / Q000096:biosynthesis* / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000378:metabolism* / Q000473:pathology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014407:Tumor Cells, Cultured
D002273:Carcinogens; D018076:DNA, Complementary; D011994:Recombinant Proteins; D019392:Cytochrome P-450 CYP2E1