Expression of Cytochromes P-450 2E1, 3A4 and 1A1/1A2 in Growing and Confluent Human HepG2 Hepatoma Cells-Effect of Ethanol

Toxicology in Vitro : an International Journal Published in Association with BIBRA
E AlexandreL Richert

Abstract

In cultured human hepatoma HepG2 cells, cytochrome (CYP) 1A-associated 7-ethoxyresorufin-O-deethylase (EROD), CYP 3A-associated benzyloxyresorufin O-debenzylase (BROD) and CYP 2E1-associated p-nitrophenol-hydroxylase (PNPH) decreased during time in culture. The enzyme activities in cells at confluence were 35-60% of the activities in cells 24 hours after seeding. Similarly, CYP 3A and CYP 2E1 proteins were present at higher concentrations in growing (G) than in confluent (C) HepG2 cells. CYP 1A1/1A2 protein was not detected, neither in G nor in C HepG2 cells but was strongly induced by 3-methylcholanthrene (3-MC) treatment. Ethanol (EtOH) was shown to increase CYP 2E1 and CYP 3A proteins and CYP 1A1/1A2-, CYP 2E1- and CYP 3A-associated mixed-function oxidase activities (MFOs) in HepG2 cells, as has been previously reported for primary cultures of human hepatocytes. These effects were observed only at the beginning of culture, in growing HepG2 cells, demonstrating the influence of the growth stage of HepG2 cells on their response to EtOH treatment. This is, to our knowledge, the first report on increases in CYP proteins and associated MFOs by EtOH in HepG2 cells. It suggests that growing HepG2 cells provide a useful in vitro mod...Continue Reading

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