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Molecular Pharmacology

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Research ArticleArticle

Dependence on the Microtubule Network and 90-kDa Heat Shock Protein of Phenobarbital-Induced Nuclear Translocation of the Rat Constitutive Androstane Receptor

Yuichiro Kanno, Yasuo Miyama, Mina Ando and Yoshio Inouye
Molecular Pharmacology February 2010, 77 (2) 311-316; DOI: https://doi.org/10.1124/mol.109.060434
Yuichiro Kanno
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Yasuo Miyama
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Mina Ando
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Yoshio Inouye
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Abstract

The role of the microtubule network in the constitutive androstane receptor (CAR)-mediated transactivation of CYP2B induced by phenobarbital (PB) in rat primary hepatocytes was investigated using the microtubule-disrupting agent nocodazole (NCZ). In human hepatocytes, it was reported that CAR mRNA expression was decreased by a microtubule-disrupting agent through the inhibition of glucocorticoid receptor (GR)-mediated transactivation. However, in the present study, we show that the rat CAR gene was unaffected by the GR-mediated pathway in rat primary hepatocytes treated with NCZ. The PB-induced expression of CYP2B mRNA was repressed in the presence of NCZ for 2 h before and during 4 h of PB treatment, whereas the CAR mRNA and protein expression levels were not affected. Furthermore, the transactivation of the PB-responsive enhancer module-luciferase reporter gene and the nuclear transport of CAR induced by PB were also repressed in the presence of NCZ. Based on these findings, microtubular integrity might be required for PB-induced nuclear translocation of CAR in rat primary hepatocytes. In the same procedures, except that NCZ was replaced with radicicol, the CYP2B mRNA expression induced by PB was also repressed. Taking these into consideration, PB-mediated nuclear translocation of rCAR might be dependent on the 90-kDa heat shock protein as well as the microtubule network.

Footnotes

  • ↵Embedded Image The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.

  • This work was partially supported by the Ministry of Education, Science, Sports, and Culture, Grant-in-Aid for Young Scientists; and by the “Open Research Center” Project.

  • Article, publication date, and citation information can be found at http://molpharm.aspetjournals.org.

    doi:10.1124/mol.109.060434

  • ABBREVIATIONS:

    CAR
    constitutive androstane receptor
    PB
    phenobarbital
    NCZ
    nocodazole
    COL
    colchicine
    RAD
    radicicol
    GR
    glucocorticoid receptor
    Hsp
    heat shock protein
    TCPOBOP
    1,4-bis[2-(3,5-dichloropyridyloxy)]benzene
    Dex
    dexamethasone
    GFP
    green fluorescent protein
    NLS
    nuclear localization signal
    PBREM
    phenobarbital-responsive enhancer module
    GA
    geldanamycin
    PCR
    polymerase chain reaction
    RT-PCR
    reverse transcription polymerase chain reaction
    DMSO
    dimethyl sulfoxide
    PBS
    phosphate-buffered saline
    PAGE
    polyacrylamide gel electrophoresis
    FK506
    tacrolimus
    SV40
    simian virus 40.

  • Received August 20, 2009.
  • Accepted November 16, 2009.
  • Copyright © 2010 The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 77 (2)
Molecular Pharmacology
Vol. 77, Issue 2
1 Feb 2010
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Research ArticleArticle

Dependence on the Microtubule Network and 90-kDa Heat Shock Protein of Phenobarbital-Induced Nuclear Translocation of the Rat Constitutive Androstane Receptor

Yuichiro Kanno, Yasuo Miyama, Mina Ando and Yoshio Inouye
Molecular Pharmacology February 1, 2010, 77 (2) 311-316; DOI: https://doi.org/10.1124/mol.109.060434

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Research ArticleArticle

Dependence on the Microtubule Network and 90-kDa Heat Shock Protein of Phenobarbital-Induced Nuclear Translocation of the Rat Constitutive Androstane Receptor

Yuichiro Kanno, Yasuo Miyama, Mina Ando and Yoshio Inouye
Molecular Pharmacology February 1, 2010, 77 (2) 311-316; DOI: https://doi.org/10.1124/mol.109.060434
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