Enzyme induction by L-buthionine (S,R)-sulfoximine in cultured mouse hepatoma cells.
Induction of Phase II enzymes of the [Ah] gene battery by L-buthionine (S,R)-sulfoximine (BSO) and other agents was examined in mouse hepatoma Hepa-1c1c7 cells. BSO, a nonelectrophilic inhibitor of gamma-glutamylcysteine synthetase (GCS), is routinely used to examine the toxicological implications of GSH depletion. Exposure to BSO for 24 h produced a 75-85% depletion of GSH levels, proportional to the inhibition of GCS activity, as well as small increases in the UDP-glucuronosyltransferase (UGT, 60%) and glutathione transferase (GST, 30%) enzyme activities in Hepa-1 wild-type (wt) cells. However, for the NAD(P)H:menadione oxidoreductase (NMO1) and cytosolic aldehyde dehydrogenase class 3 (AHD4) enzyme activities, BSO produced larger increases (110% and 170%, respectively). The mechanisms of NMO1 and AHD4 induction were examined further. In Hepa-1 wt cells, NMO1 and AHD4 activities were increased by the aromatic hydrocarbon inducer 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and by the electrophile tert-butylhydroquinone (tBHQ), known inducing agents for these enzymes. However, NMO1 and AHD4 were induced in Ah receptor nuclear translocation-defective mutant (c4) cells by BSO and tBHQ, but not by TCDD, suggesting that the induction by BSO and tBHQ is not Ah receptor-mediated. In wt cells, N-acetylcysteine produced a concentration-dependent increase in intracellular cysteine levels, but not GSH levels, in the absence or presence of BSO. Furthermore, N-acetylcysteine had no effect on NMO1 activity under any conditions examined, suggesting that GSH levels per se, rather than change in overall thiol status, might be mediating increased NMO1 activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Shertzer
H G
HG
Department of Environmental Health, University of Cincinnati Medical Center, Ohio 45267-0056, USA.
Vasiliou
V
V
Liu
R M
RM
Tabor
M W
MW
Nebert
D W
DW
D019881:Aminoacyltransferases; D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D015152:Blotting, Northern; D019328:Buthionine Sulfoximine; D002460:Cell Line; D003609:Dactinomycin / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006873:Hydroquinones / Q000494:pharmacology; D008114:Liver Neoplasms, Experimental; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives* / Q000494:pharmacology; D051379:Mice; D010458:Peptidyl Transferases / Q000096:biosynthesis* / Q000187:drug effects; D013438:Sulfhydryl Compounds / Q000378:metabolism
D000963:Antimetabolites; D000975:Antioxidants; D006873:Hydroquinones; D013438:Sulfhydryl Compounds; D008717:Methionine Sulfoximine; D003609:Dactinomycin; D019328:Buthionine Sulfoximine; C018855:2-tert-butylhydroquinone; D019881:Aminoacyltransferases; D010458:Peptidyl Transferases; C093784:glutathione gamma-glutamylcysteinyltransferase