Nuclear uptake of the Ah (dioxin) receptor in response to omeprazole: transcriptional activation of the human CYP1A1 gene.
In the presence of halogenated and polycyclic aromatic hydrocarbons, the CYP1A1 gene is regulated through induction after ligand binding to the cytosolic Ah receptor (AhR). Ligand-dependent AhR activation leads to nuclear translocation and binding of the receptor to dioxin-responsive element (DRE) sequences, an event that initiates transcriptional activation of the CYP1A1 gene. We recently established a human hepatoma cell line stably integrated with the human CYP1A1 promoter and 5'-flanking enhancer sequences fused to the firefly luciferase gene. This cell line, 101L, was used to determine whether the induction of CYP1A1 by omeprazole, a gastric proton pump inhibitor, is AhR mediated. Treatment of 101L cells with either 50 microM omeprazole or 5 nM 2,3,7,8-tetrachlorodibenzo-p-dioxin for 12-72 hr resulted in maximal activity at 24 hr for both inducers. A dose-response curve for omeprazole induction at 24 hr was determined and the EC50 for omeprazole induction of the human CYP1A1 gene was estimated to be 100 microM. The induction of the CYP1A1 gene by omeprazole corresponds to increases in CYP1A1 mRNA. To examine whether omeprazole-initiated transcriptional activation of the CYP1A1 gene correlates with nuclear accumulation of the AhR, binding of nuclear proteins to the DRE was examined. When gel mobility shift assays were performed using nuclear extracts isolated from 101L cells treated with omeprazole or 2,3,7,8-tetrachlorodibenzo-p-dioxin, specific binding of the AhR to the DRE was observed. These studies demonstrate that omeprazole initiates AhR activation and that induction of the human CYP1A1 gene by omeprazole is AhR dependent.
Department of Pharmacology, UCSD Cancer Center, La Jolla 92093.
L C
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Quattrochi
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Tukey
D001483:Base Sequence; D002467:Cell Nucleus / Q000187:drug effects* / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000187:drug effects* / Q000235:genetics; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009853:Omeprazole / Q000494:pharmacology*; D000072317:Polychlorinated Dibenzodioxins / Q000494:pharmacology; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug / Q000187:drug effects* / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
D000072317:Polychlorinated Dibenzodioxins; D018336:Receptors, Aryl Hydrocarbon; D011955:Receptors, Drug; D003577:Cytochrome P-450 Enzyme System; D008156:Luciferases; D009853:Omeprazole