The effect of ethanol and nitric oxide on the N-nitrosodimethylamine formation in HepG2 cells overexpressing CYP2E1.
The influence of lipopolysaccharide (LPS) and the nitric oxide synthase (iNOS) inhibitor--N-nitro-L-arginine methyl ester (L-NAME)--on the formation of N-nitrosodimethylamine (NDMA) by HepG2 cells, engineered to overexpress CYP2E1, was assessed and compared with data from empty vector-transfected cells. HepG2 cells produced significant amounts of NDMA but its levels in the culture media of cells overexpressing CYP2E1 was significantly lower than in empty-vector transfected cells. LPS increased the formation of NDMA, the expression of the iNOS and the production of the nitric oxide (NO). On the other hand, L-NAME significantly decreased NDMA levels. The results above indicate that the synthesis of NDMA by HepG2 cells depends on NO production. Furthermore, ethanol did not affect iNOS expression but decreased NDMA levels in CYP2E1-transfected cells below the detection limit. It is probably caused by the increased N-nitrosodimethylamine metabolism. In conclusion, HepG2 cells' ability to synthesize NO with simultaneous CYP2E1 activation may lead to an increase of carcinogenic products of the NDMA metabolism.
Jabloński
Jakub
J
Department of Toxicology, Medical University of Bialystok, Bialystok, Poland. toksykologia@wp.pl
Hołownia
Adam
A
Jabłońska
Ewa
E
Moniuszko-Jakoniuk
Janina
J
Braszko
Jan
J
Iwanowska
Jolanta
J
Marcińczyk
Magda
M
D045744:Cell Line, Tumor; D003470:Culture Media / Q000032:analysis; D019392:Cytochrome P-450 CYP2E1 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004128:Dimethylnitrosamine; D000431:Ethanol / Q000494:pharmacology*; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009566:Nitrates / Q000032:analysis; D009569:Nitric Oxide / Q000494:pharmacology*; D009573:Nitrites / Q000032:analysis; D009602:Nitrosamines / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
D003470:Culture Media; D008070:Lipopolysaccharides; D009566:Nitrates; D009573:Nitrites; D009602:Nitrosamines; D009569:Nitric Oxide; D000431:Ethanol; D019392:Cytochrome P-450 CYP2E1; D004128:Dimethylnitrosamine; D019331:NG-Nitroarginine Methyl Ester