Induction of cytochrome P450-dependent monooxygenase in serum-free cultured Hep G2 cells.
We examined the induction of cytochrome P450-dependent mixed-function monooxygenase (MFO) in the human hepatoma cell line Hep G2 by means of several factors. The MFO activities induced in the cells cultured in medium containing five commercial sera varied significantly, and the activity in the cells cultured in the absence of serum was about twice as high as that in cells supplemented with serum. The activity of ethoxycoumarin O-deethylase was highest 12 hr after adding 3-methylcholanthrene, and it was induced by several polycyclic aromatic hydrocarbons such as benzo(a)anthracene and benzo(a)pyrene, which are usually found in urban air as environmental contaminants. Furthermore, an extract from the total suspended particles collected using a high volume air sampler, which was mutagenic in the Ames assay using Salmonella typhimurium TA98, induced the same enzyme activities in Hep G2 cells. These findings suggest that serum-free culture allows the stable and highly sensitive measurement of induced MFO activity, and that studies of MFO induction by environmental samples using human hepatoma Hep G2 cells should provide helpful information regarding the risk associated with environmental contaminants.
Nakama
A
A
Osaka City Institute of Public Health and Environmental Sciences, Japan.
Kuroda
K
K
Yamada
A
A
D015660:7-Alkoxycoumarin O-Dealkylase / Q000096:biosynthesis* / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D016895:Culture Media, Serum-Free; D019363:Cytochrome P-450 CYP1A1; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000378:metabolism; D004790:Enzyme Induction; D006801:Humans; D006843:Hydrocarbons, Chlorinated / Q000633:toxicity; D008113:Liver Neoplasms / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000096:biosynthesis* / Q000378:metabolism; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity; D010088:Oxidoreductases / Q000096:biosynthesis* / Q000378:metabolism; D011083:Polycyclic Compounds / Q000633:toxicity; D014407:Tumor Cells, Cultured
D016895:Culture Media, Serum-Free; D006843:Hydrocarbons, Chlorinated; D009153:Mutagens; D011083:Polycyclic Compounds; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; D015660:7-Alkoxycoumarin O-Dealkylase; D019363:Cytochrome P-450 CYP1A1