title: Epidermal Growth Factor Receptor Kinase Inhibitors Synergize with TCDD to Induce CYP1A1/1A2 in Human Breast Epithelial MCF10A Cells.
abstract: CYP1A1 and CYP1A2 are transcriptionally activated in the human normal breast epithelial cell line MCF10A following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Shifting MCF10A cultures to medium deficient in serum and epidermal growth factor (EGF) caused rapid reductions in the activated (i.e., phosphorylated) forms of extracellular regulated kinases (ERKs) and the epidermal growth factor receptor (EGFR). Shifting to serum/EGF-deficient medium also enhanced TCDD-mediated induction of cytochrome P450 (CYP)1A1 Treatment of cells cultured in complete medium with the EGFR inhibitors gefitinib (Iressa), AG1478, and CI-1033 resulted in concentration-dependent reductions of active EGFR and ERKs, and increased CYP1A1 mRNA content ∼3- to 18-fold above basal level. EGFR inhibitors synergized with TCDD and resulted in transient CYP1A1 and CYP1A2 mRNA accumulations ∼8-fold greater (maximum at 5 hours) than that achieved with only TCDD. AG1478, gefitinib, and TCDD individually induced small increases (∼1.2- to 2.5-fold) in CYP1A1 protein content but did not cause additive or synergistic accumulations of CYP1A1 protein when used in combination. The mitogen-activated protein kinase kinase inhibitor PD184352 inhibited ERK and EGFR activation in a concentration-dependent fashion without causing CYP1A1 mRNA accumulation. However, cotreatment with PD184352 potentiated TCDD-mediated CYP1A1 induction. TCDD-mediated induction of CYP1A1 in MCF7-TET on-EGFR cells, a MCF7 variant in which EGFR expression can be controlled, was not affected by the activity status of EGFR or ERKs. Hence, EGFR signaling mutes both basal and ligand-induced expression of two aryl hydrocarbon receptor-responsive P450s in MCF10A cultures. However, these effects are cell context-dependent. Furthermore, CYP1A1 mRNA and protein abundance are not closely coupled in MCF10A cultures.
authors.affiliation: Institute of Environmental Health Sciences, Wayne State University, Detroit, Michigan (A.J., P.A.M., J.J.R.); Department of Pharmacology (C.S., J.J.R.) and Department of Oncology (J.B.), Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Aby
authors.initials: A
authors.lastname: Joiakim
authors.affiliation: Institute of Environmental Health Sciences, Wayne State University, Detroit, Michigan (A.J., P.A.M., J.J.R.); Department of Pharmacology (C.S., J.J.R.) and Department of Oncology (J.B.), Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Mathieu
authors.affiliation: Institute of Environmental Health Sciences, Wayne State University, Detroit, Michigan (A.J., P.A.M., J.J.R.); Department of Pharmacology (C.S., J.J.R.) and Department of Oncology (J.B.), Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Catherine
authors.initials: C
authors.lastname: Shelp
authors.affiliation: Institute of Environmental Health Sciences, Wayne State University, Detroit, Michigan (A.J., P.A.M., J.J.R.); Department of Pharmacology (C.S., J.J.R.) and Department of Oncology (J.B.), Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan.
authors.forename: Julie
authors.initials: J
authors.lastname: Boerner
authors.affiliation: Institute of Environmental Health Sciences, Wayne State University, Detroit, Michigan (A.J., P.A.M., J.J.R.); Department of Pharmacology (C.S., J.J.R.) and Department of Oncology (J.B.), Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan john.reiners.jr@wayne.edu.
authors.forename: John J
authors.initials: JJ
authors.lastname: Reiners
mesh_terms: D001549:Benzamides / Q000494:pharmacology; D001940:Breast / Q000187:drug effects* / Q000378:metabolism; D002460:Cell Line; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism*; D019388:Cytochrome P-450 CYP1A2 / Q000378:metabolism*; D004357:Drug Synergism; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D000077156:Gefitinib; D006801:Humans; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors; D009025:Morpholines / Q000494:pharmacology; D000072317:Polychlorinated Dibenzodioxins / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020032:Tyrphostins / Q000494:pharmacology
chemical_list: C120227:2-(2-chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide; D001549:Benzamides; D009025:Morpholines; D000072317:Polychlorinated Dibenzodioxins; D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D012333:RNA, Messenger; D020032:Tyrphostins; C101044:RTKI cpd; C420268:Canertinib; C569463:CYP1A1 protein, human; C504573:CYP1A2 protein, human; D019363:Cytochrome P-450 CYP1A1; D019388:Cytochrome P-450 CYP1A2; D066246:ErbB Receptors; D048369:MAP Kinase Kinase 1; D000077156:Gefitinib