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Research ArticleABSORPTION, DISTRIBUTION, METABOLISM, AND EXCRETION

Induction of Cytochrome P-450 1A2 by Oxidized Tryptophan in Hepa lclc7 Cells

Ram K. Sindhu, Masato Mitsuhashi and Yutaka Kikkawa
Journal of Pharmacology and Experimental Therapeutics March 2000, 292 (3) 1008-1014;
Ram K. Sindhu
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Masato Mitsuhashi
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Yutaka Kikkawa
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Abstract

Recent studies from this laboratory have demonstrated thatl-tryptophan, after oxidation either by UV-irradiation or ozone, induces aryl hydrocarbon receptor (AhR) activation and binding of the liganded AhR complex to its specific DNA recognition site, thereby initiating transcription of the cytochrome P-450 1a1 (Cyp1a1) gene with concomitant increase of CYP1A1 protein and 7-ethoxyresorufin O-deethylase activity in wild-type mouse hepatoma cells, Hepa lclc7 (Hepa-1), in culture. Temporary inhibition of protein synthesis by cycloheximide resulted in superinduction of oxidized tryptophan-inducible CYP1A1 mRNA, protein, and 7-ethoxyresorufin O-deethylase activity in Hepa-1 cells. In the present communication, the results obtained by immunoblot analyses with monoclonal CYP1A1/1A2 antibody (NIH 1-7-1) demonstrate that both UV- or ozone-oxidized tryptophan also induce CYP1A2 protein in Hepa-1 cells. CYP1A2 mRNA, detected by reverse transcription-polymerase chain reaction, was markedly induced in the UV- or ozone-oxidized tryptophan-treated cells. Temporary inhibition of protein synthesis by cycloheximide further induced oxidized tryptophan-inducible CYP1A2 mRNA as well as the protein in Hepa-1 cells. This is the first report demonstrating the induction of CYP1A2 mRNA and protein in Hepa-1 cells.

Footnotes

  • Send reprint requests to: Ram K. Sindhu, Ph.D., Department of Pathology, College of Medicine, University of California-Irvine, Irvine, CA 92697-4800. E-mail: rksindhu{at}uci.edu

  • ↵1 This work was supported by Grant HL-50450 from the National Heart, Lung and Blood Institute of the National Institutes of Health, U.S. Public Health Service.

  • Abbreviations:
    CYP
    cytochrome P-450
    PAH
    polycyclic aromatic hydrocarbon
    Ah
    aryl hydrocarbon
    OT
    ozone-oxidized tryptophan
    UVT
    UV-oxidized tryptophan
    AhR
    aryl hydrocarbon receptor
    EROD
    7-ethoxyresorufin O-deethylase
    TCDD
    2,3,7,8-tetrachlorodibenzo-p-dioxin
    PAGE
    polyacrylamide gel electrophoresis
    RT-PCR
    reverse transcription-polymerase chain reaction
    bp
    base pair
    MC
    3-methylcholanthrene
    • Received July 7, 1999.
    • Accepted November 18, 1999.
  • The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 292 (3)
Journal of Pharmacology and Experimental Therapeutics
Vol. 292, Issue 3
1 Mar 2000
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Research ArticleABSORPTION, DISTRIBUTION, METABOLISM, AND EXCRETION

Induction of Cytochrome P-450 1A2 by Oxidized Tryptophan in Hepa lclc7 Cells

Ram K. Sindhu, Masato Mitsuhashi and Yutaka Kikkawa
Journal of Pharmacology and Experimental Therapeutics March 1, 2000, 292 (3) 1008-1014;

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Research ArticleABSORPTION, DISTRIBUTION, METABOLISM, AND EXCRETION

Induction of Cytochrome P-450 1A2 by Oxidized Tryptophan in Hepa lclc7 Cells

Ram K. Sindhu, Masato Mitsuhashi and Yutaka Kikkawa
Journal of Pharmacology and Experimental Therapeutics March 1, 2000, 292 (3) 1008-1014;
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