title: Stabilization of cytochrome P4502E1 protein by ethanol in primary hamster hepatocyte cultures.
abstract: We report the effect of ethanol on monooxygenase activities in primary hamster hepatocyte cultures maintained on collagen-coated dishes. The addition of 50mM ethanol to cell cultures both from control and ethanol pretreated animals almost completely maintained, at least for 72hr, the P4502E1-dependent aniline hydroxylase (AnH) activity and the 2E1 immunodetectable apoprotein content at the levels of the corresponding 4-hr plated hepatocytes. On the contrary, other P450-dependent monooxygenase activities, as assayed by testosterone hydroxylases, kept decreasing falling-after 72hr of culture-to the levels of the 4-hr plated hepatocytes. In both cases, in the absence of ethanol, a rapid decline of AnH activities and 2E1 apoprotein contents were also observed, attaining undetectable levels at 72hr. The hybridizable 2E1 mRNA also rapidly declined in both cultures, but such decline was not significantly altered by the presence of 50mM ethanol in the culture medium. Furthermore, we show that P4502E1 in the liver possesses a rapid degradation phase with a half-life of about 6hr. Thus, in the hamster, P4502E1 appears regulated at post-translational level, as in rat, probably by a protein stabilization mechanism.
authors.affiliation: Laboratory of Genetic and Biochemical Toxicology, Istituto di Mutagenesi e Differenziamento CNR, via Svezia 10, 56124 Pisa, Italy.
authors.forename: U
authors.initials: U
authors.lastname: Zanelli
authors.forename: V
authors.initials: V
authors.lastname: Longo
authors.forename: A
authors.initials: A
authors.lastname: Paolicchi
authors.forename: P G
authors.initials: PG
authors.lastname: Gervasi
mesh_terms: D000815:Aniline Hydroxylase / Q000378:metabolism; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D015152:Blotting, Northern; D002469:Cell Separation; D002478:Cells, Cultured; D006224:Cricetinae; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D019392:Cytochrome P-450 CYP2E1 / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology*; D008099:Liver / Q000166:cytology* / Q000187:drug effects; D008297:Male; D008647:Mesocricetus; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D013250:Steroid Hydroxylases / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D019392:Cytochrome P-450 CYP2E1; D000815:Aniline Hydroxylase; D001189:Aryl Hydrocarbon Hydroxylases; D019363:Cytochrome P-450 CYP1A1; C022724:testosterone 7-alpha-hydroxylase, hamster