Effect of N,N'-dinitrosopiperazine on in vitro expression of human cytochrome P450 2E1.
OBJECTIVE
To establish an in vitro heterogeneous expression model of human CYP2E1 (hCYP2E1) cDNA and investigate the effect of the chemical carcinogenic N, N'-dinitrosopiperazine (DNP) on the expression of CYP2E1.


METHODS
Exogenous hCYP2E1 was introduced into the mouse derived NIH3T3 cells using the lipofectamine transfection technique. Integration of exogenous hCYP2E1 gene was identified by PCR and Southern blot. After treatment with various concentration of ethanol and DNP on the transfected NIH3T3 cell cultures, RT-PCR and Western blot was applied to detect the expression level of CYP2E1.


RESULTS
Two cell clones with integration and stable expression of exogenous hCYP2E1 were obtained and designated as NIH3T3-2E1-A4 and NIH3T3-2E1-A8 respectively. The expression of both hCYP2E1 mRNA and protein products was promoted after either ethanol or DNP treatment.


CONCLUSIONS
The results suggested that the promoted expression of hCYP2E1 induced by DNP and /or ethanol is due to enhanced transcription. The mechanism of DNP carcinogenes is might be related to this in situ activated metabolism by CYP2E1.
Zhu
Jianhua
J
Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha 410078, China.
He
Zhimin
Z
Wang
Shuiliang
S
Chen
Zhuchu
Z
D016475:3T3 Cells; D000818:Animals; D002273:Carcinogens / Q000633:toxicity*; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics*; D000431:Ethanol / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D006801:Humans; D051379:Mice; D009602:Nitrosamines / Q000633:toxicity*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D002273:Carcinogens; D009602:Nitrosamines; D000431:Ethanol; D019392:Cytochrome P-450 CYP2E1; C026156:N,N'-dinitrosopiperazine