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Research ArticleMetabolism, Transport, and Pharmacogenomics

Regulation of Estrogen Sulfotransferase Expression by Confluence of MCF10A Breast Epithelial Cells: Role of the Aryl Hydrocarbon Receptor

Jiaqi Fu, Hailin Fang, Michelle Paulsen, Mats Ljungman, Thomas A. Kocarek and Melissa Runge-Morris
Journal of Pharmacology and Experimental Therapeutics November 2011, 339 (2) 597-606; DOI: https://doi.org/10.1124/jpet.111.185173
Jiaqi Fu
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Hailin Fang
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Michelle Paulsen
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Mats Ljungman
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Thomas A. Kocarek
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Melissa Runge-Morris
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Abstract

Estrogen sulfotransferase (SULT1E1) catalyzes the sulfonation of estrogens, which limits estrogen mitogenicity. We recently reported that SULT1E1 expression is low in preconfluent MCF10A human breast epithelial cells but increases when the cells become confluent. Pulse-chase labeling experiments with 5-bromouridine demonstrated that the confluence-mediated increase in SULT1E1 expression was due to increased mRNA synthesis. Because aryl hydrocarbon receptor (AhR) activation has been shown to suppress SULT1E1 expression and loss of cell-cell contact has been shown to activate the AhR in other cell types, we tested whether the confluence-associated changes in SULT1E1 expression were mediated by the AhR. Relative to confluent MCF10A cells, preconfluent cells had higher levels of CYP1A1 mRNA and greater activation of an AhR-responsive luciferase reporter, demonstrating that the AhR was active in the preconfluent cells. AhR and aryl hydrocarbon receptor nuclear translocator mRNA and protein levels were also higher in preconfluent than in confluent cultures. Treatment of preconfluent cells with the AhR antagonist, 3′-methoxy-4′-nitroflavone (MNF), or AhR knockdown significantly increased SULT1E1 expression. MCF10A cells stably transfected with a luciferase reporter containing ∼7 kilobases of the SULT1E1 5′-flanking region showed both MNF- and confluence-inducible luciferase expression. Preconfluent cells transiently transfected with the reporter showed both MNF treatment- and AhR knockdown-mediated luciferase induction, but mutation of a computationally predicted dioxin response element (DRE) at nucleotide (nt) −3476 did not attenuate these effects. These results demonstrate that SULT1E1 expression in MCF10A cells is transcriptionally regulated by confluence through a suppressive action of the AhR, which is not mediated through a DRE at nt −3476.

Footnotes

  • This work was supported by the National Institutes of Health National Institute of Environmental Health Sciences [Grant ES05823]; and Uniting Against Lung Cancer.

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

    doi:10.1124/jpet.111.185173.

  • ABBREVIATIONS:

    SULT
    cytosolic sulfotransferase
    SULT1E1
    estrogen sulfotransferase
    ER
    estrogen receptor
    AhR
    aryl hydrocarbon receptor
    TCDD
    2,3,7,8-tetrachlorodibenzo-p-dioxin
    MNF
    3′-methoxy-4′-nitroflavone
    BrU
    5-bromouridine
    GAPDH
    glyceraldehyde-3-phosphate dehydrogenase
    DMSO
    dimethyl sulfoxide
    ARNT
    aryl hydrocarbon receptor nuclear translocator
    PCR
    polymerase chain reaction
    hnRNA
    heterogeneous nuclear RNA
    RT
    reverse transcriptase
    nt
    nucleotide
    PBS
    phosphate-buffered saline
    DRE
    dioxin response element
    kb
    kilobase
    Ct
    cycle threshold.

  • Received June 17, 2011.
  • Accepted August 8, 2011.
  • Copyright © 2011 by The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 339 (2)
Journal of Pharmacology and Experimental Therapeutics
Vol. 339, Issue 2
1 Nov 2011
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Research ArticleMetabolism, Transport, and Pharmacogenomics

SULT1E1 Regulation by Confluence of MCF10A Cells

Jiaqi Fu, Hailin Fang, Michelle Paulsen, Mats Ljungman, Thomas A. Kocarek and Melissa Runge-Morris
Journal of Pharmacology and Experimental Therapeutics November 1, 2011, 339 (2) 597-606; DOI: https://doi.org/10.1124/jpet.111.185173

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Research ArticleMetabolism, Transport, and Pharmacogenomics

SULT1E1 Regulation by Confluence of MCF10A Cells

Jiaqi Fu, Hailin Fang, Michelle Paulsen, Mats Ljungman, Thomas A. Kocarek and Melissa Runge-Morris
Journal of Pharmacology and Experimental Therapeutics November 1, 2011, 339 (2) 597-606; DOI: https://doi.org/10.1124/jpet.111.185173
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