title: Bone marrow stromal cells constitutively express high levels of cytochrome P4501B1 that metabolize 7,12-dimethylbenz[a]anthracene.
abstract: The polycyclic aromatic hydrocarbon 7,12-dimethylbenz[a]anthracene (DMBA) is a potent carcinogen that produces immunotoxic effects in bone marrow. Here, we show that bone marrow stromal cells metabolize DMBA to such products as 3,4-dihydrodiol, the precursor to the most mutagenic DMBA metabolite. The BMS2 bone marrow stromal cell line constitutively expressed higher levels of CYP1B1 protein and mRNA than C3H10T1/2 mouse embryo fibroblasts. BMS2 cells also produced a DMBA metabolite profile that was consistent with CYP1B1 activity. Treatment with the potent aryl hydrocarbon receptor (AhR) ligand 2,3, 7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced a approximately 2-fold increase in CYP1B1 mRNA, protein, and activity in BMS2 cells. Two forms of the AhR (97 and 104 kDa) and the AhR nuclear translocator were detected in BMS2 cells. The AhR translocated to the nucleus after treatment with TCDD or DMBA but was approximately 5 times slower with DMBA. Primary bone marrow stromal (BMS) cell cultures established from AhR-/- mice showed similar basal CYP1B1 expression and activity as cell cultures established from heterozygous littermates or C57BL/6 mice. However, primary BMS cells from AhR-/- mice did not exhibit increased CYP1B1 protein expression after incubation with TCDD. BMS cells therefore constitutively express functional CYP1B1 that is not dependent on the AhR. This contrasts with embryo fibroblasts from the same mouse strain, in which basal CYP1B1 expression is AhR dependent. We therefore conclude that bone marrow toxicity may be mediated by CYP1B1-dependent DMBA metabolism, which is regulated by factors other than the AhR.
authors.affiliation: The Center for Environmental Toxicology, University of Wisconsin, Madison, Wisconsin 53706, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Heidel
authors.forename: C J
authors.initials: CJ
authors.lastname: Czuprynski
authors.forename: C R
authors.initials: CR
authors.lastname: Jefcoate
mesh_terms: D015127:9,10-Dimethyl-1,2-benzanthracene / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001854:Bone Marrow Cells / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism*; D002478:Cells, Cultured; D065633:Cytochrome P-450 CYP1B1; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000378:metabolism*; D004789:Enzyme Activation; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity; D018336:Receptors, Aryl Hydrocarbon / Q000172:deficiency / Q000378:metabolism
chemical_list: D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D015127:9,10-Dimethyl-1,2-benzanthracene; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C585498:Cyp1b1 protein, mouse; D065633:Cytochrome P-450 CYP1B1