Application of primary hepatocytes from p53-knockout mice for studies of expression of Cyp3a.
CYP3A rapidly disappears in primary hepatocytes, although the primary cells are suitable for studies of the regulation of CYP3A genes. In the present study, we found that Cyp3a mRNA could be expressed in the primary hepatocytes from p53-knockout mice for at least 2 weeks when the cells were cultured in the presence of dexamethasone. Propoxycoumarin O-depropylase activity, which is known to be mainly catalyzed by CYP3A, was maintained at a level of 50% of the initial activity even after 5 days of culture, and the activity correlated with the expression level of Cyp3a mRNA in the primary hepatocytes from p53-knockout mice. The cells remained morphologically intact during 4 weeks. These results suggest that hepatocytes from p53-knockout mice are a useful tool for studies of the expression of Cyp3a.
Shimoji
M
M
Division of Drug Metabolism, Faculty of Pharmaceutical Sciences, Hokkaido University.
Itoh
S
S
Toide
K
K
Nakayama
K
K
Aizawa
S
S
Kamataki
T
T
D015660:7-Alkoxycoumarin O-Dealkylase / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D002455:Cell Division; D002478:Cells, Cultured; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D003907:Dexamethasone / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010089:Oxidoreductases, N-Demethylating / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
D007328:Insulin; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D004815:Epidermal Growth Factor; D017228:Hepatocyte Growth Factor; D003907:Dexamethasone; D003577:Cytochrome P-450 Enzyme System; D015660:7-Alkoxycoumarin O-Dealkylase; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; D010089:Oxidoreductases, N-Demethylating