Bromobenzene detoxification in the human liver-derived HepG2 cell line.
1. The applicability of the human hepatoma cell line, HepG2, as a cell culture model for studying xenobiotic liver toxicity has been investigated using the well-characterized hepatotoxic chemical, bromobenzene. 2. Bromobenzene caused a concentration- (0-10 mM) and time-dependent (0-180 min) decrease in HepG2 cell viability. The degree of toxicity was dependent upon the culture medium composition and the state of cell growth. Toxicity in Modified Earle's and Williams' E Media was maximal at 7 days growth compared with 3 and 10 days, and was greater in Williams' than in Earle's medium. Toxicity in Dulbecco's medium was apparent only at 10 days growth and was less than the maximum toxicity in the other media. 3. Bromobenzene was detoxified by epoxide hydrase. The question of metabolic activation by P450 remained unresolved, but any involvement of P450 was by forms not inhibited by ketoconazole. 4. The mechanism of bromobenzene toxicity did not appear to involve lipid peroxidation, depletion of reduced glutathione, calcium-mediated proteolysis or metabolic activation by prostaglandin synthetase, but may have involved direct solvent-induced cell damage. 5. This study demonstrates the potential usefulness of HepG2 cells in toxicity testing and highlights the importance of standardizing culture conditions.
Duthie
S J
SJ
Department of Biomedical Sciences, University of Aberdeen, Marischal College, UK.
Melvin
W T
WT
Burke
M D
MD
D000976:Antipain / Q000494:pharmacology; D001969:Bromobenzenes / Q000378:metabolism* / Q000493:pharmacokinetics / Q000633:toxicity; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004851:Epoxide Hydrolases / Q000378:metabolism; D005609:Free Radicals; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008658:Inactivation, Metabolic; D015227:Lipid Peroxidation; D008099:Liver / Q000378:metabolism*; D008113:Liver Neoplasms / Q000378:metabolism; D008954:Models, Biological; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014810:Vitamin E / Q000494:pharmacology
D001969:Bromobenzenes; D003470:Culture Media; D005609:Free Radicals; D014810:Vitamin E; D000976:Antipain; D003577:Cytochrome P-450 Enzyme System; C032036:bromobenzene; D011451:Prostaglandin-Endoperoxide Synthases; D004851:Epoxide Hydrolases; D005978:Glutathione