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Research ArticleArticle

Transcriptional Regulation of CYP2B1 Induction in Primary Rat Hepatocyte Cultures: Repression by Epidermal Growth Factor Is Mediated via a Distal Enhancer Region

Daniel Bauer, Nils Wolfram, Georg F. Kahl and Karen I. Hirsch-Ernst
Molecular Pharmacology January 2004, 65 (1) 172-180; DOI: https://doi.org/10.1124/mol.65.1.172
Daniel Bauer
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Nils Wolfram
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Georg F. Kahl
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Karen I. Hirsch-Ernst
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Abstract

Phenobarbital (PB) alters expression of numerous hepatic genes, including genes involved in xenobiotic metabolism. Phenobarbital-dependent induction of cytochrome P-450 2B1 (CYP2B1) is subject to regulation by cytokines [e.g., by epidermal growth factor (EGF)], hormones [e.g., by growth hormone (GH)], or the cellular redox status. To investigate mechanisms involved in regulation of CYP2B1 transcription, we performed promoter activation studies using primary rat hepatocyte cultures transiently transfected with individual CYP2B1 promoter-luciferase reporter gene constructs. The 2679-bp native 5′-flanking region of the CYP2B1 gene conferred reporter gene activation by PB and the potent PB-like inducer permethrin (PM). Furthermore, this region mediated EGF- and GH-dependent repression of gene activation by PB-like inducers. A wide promoter mapping strategy with constructs bearing internal CYP2B1 promoter deletions led to identification of a distal responsive CYP2B1 enhancer region at -2230 to -2170, encompassing the section equivalent to the 51-bp PB-responsive enhancer module situated in the distal mouse Cyp2b10-5′-flanking region. The distal CYP2B1 enhancer region conferred gene activation by PM, repression of PM-dependent activation by EGF, and enhancement of activation by the antioxidant N-acetylcysteine (NAC). Mutational analyses of the region at -2230 to -2170 suggested that the mechanisms of PB-dependent induction of CYP2B1 and the modulating effects by EGF or NAC are closely related.

  • Received June 27, 2003.
  • Accepted October 6, 2003.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 65 (1)
Molecular Pharmacology
Vol. 65, Issue 1
1 Jan 2004
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Research ArticleArticle

Transcriptional Regulation of CYP2B1 Induction in Primary Rat Hepatocyte Cultures: Repression by Epidermal Growth Factor Is Mediated via a Distal Enhancer Region

Daniel Bauer, Nils Wolfram, Georg F. Kahl and Karen I. Hirsch-Ernst
Molecular Pharmacology January 1, 2004, 65 (1) 172-180; DOI: https://doi.org/10.1124/mol.65.1.172

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Research ArticleArticle

Transcriptional Regulation of CYP2B1 Induction in Primary Rat Hepatocyte Cultures: Repression by Epidermal Growth Factor Is Mediated via a Distal Enhancer Region

Daniel Bauer, Nils Wolfram, Georg F. Kahl and Karen I. Hirsch-Ernst
Molecular Pharmacology January 1, 2004, 65 (1) 172-180; DOI: https://doi.org/10.1124/mol.65.1.172
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