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The rexinoid bexarotene represses cyclin D1 transcription by inducing the DEC2 transcriptional repressor

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Abstract

Bexarotene is an RXR-selective vitamin A analog that has been shown to prevent ER-negative mammary tumorigenesis in animal models. While investigating the mechanism by which bexarotene prevents ER-negative breast cancer development, we found that the expression of cyclin D1, a critical cell cycle promoter, was repressed by bexarotene in vitro and in vivo. Time course and cycloheximide experiments show that repression of cyclin D1 is a late effect and requires new protein synthesis. Previously we discovered that DEC2 (differentially expressed in chondrocytes-2), a helix–loop–helix transcription repressor, was induced by bexarotene in human mammary epithelial cells. Therefore, we hypothesized that bexarotene represses the transcription of cyclin D1 through induction of DEC2. Luciferase reporter studies demonstrated that either bexarotene treatment or forced expression of DEC2 can repress the transcription of a cyclin D1 promoter reporter by affecting the basal transcriptional activity. Results from chromatin immunoprecipitation experiments showed that bexarotene treatment causes the recruitment of DEC2 and HDAC1 (histone deacetylase 1) to the cyclin D1 promoter. Co-immunoprecipitation confirms the interaction between DEC2 and HDAC1, suggesting that the recruitment of HDAC1 to the cyclin D1 promoter is through DEC2. Trichostatin A, a HDAC inhibitor, reverses the cyclin D1 repression by bexarotene, suggesting that repression of cyclin D1 involves histone deacetylation. Knock-down of DEC2 by siRNA abolishes the cyclin D1 repression, further supporting our hypothesis. Finally, we demonstrated that overexpression of DEC2 dramatically inhibited cell proliferation and repressed the expression of cyclin D1 in human mammary epithelial cells. These results suggest that bexarotene down-regulates cyclin D1 through induction of DEC2, followed by recruitment of HDAC1 to the cyclin D1 promoter causing transcriptional repression. By elucidating the mechanism by which rexinoids inhibit cell proliferation, it will be possible to develop more effective and less toxic drugs to prevent ER-negative breast cancers.

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Abbreviations

HMEC:

Human mammary epithelial cells

RAR:

Retinoic acid receptor

RXR:

Retinoid X receptor

RT-PCR:

Reverse transcriptase PCR

ER:

Estrogen receptor

SERM:

Selective estrogen receptor modulator

DEC2:

Differentially expressed in chondrocyte 2

HDAC1:

Histone deacetylase 1

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Acknowledgments

We thank Dr. Julie Mayer for critical review and helpful discussion of the manuscript. We thank Kimberly Young-Jenkins for her assistance in submitting this manuscript. This work was supported by: NIH RO1 CA078480 and a Postdoctoral Fellowship from The Susan G. Komen Breast Cancer Foundation to Yuxin Li.

Conflict of interest

Reid P. Bissonnette is employed in the Department of Retinoid Research, Ligand Pharmaceuticals, Inc., San Diego, CA 92121. W. W. Lamph is a previous employee of Ligand Pharmaceuticals. Powel H. Brown serves as a Scientific Advisory Board member for Susan G. for the Cure, not for profit organization. All other authors have no conflict of interest.

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Authors and Affiliations

  1. Department of Clinical Cancer Prevention, The University of Texas, Anderson Cancer Center Cancer, Prevention Building (CPB6. 3468) 1515 Holcombe Blvd., Unit 1360, Houston, TX, 77030, USA

    Yuxin Li, Qiang Shen, Hee-Tae Kim & Powel H. Brown

  2. Department of Retinoid Research, Ligand Pharmaceuticals Inc, San Diego, CA, 92121, USA

    Reid P. Bissonnette

  3. San Diego, CA, USA

    William W. Lamph

  4. Department of Biomedical and Pharmaceutical Sciences, University of Rhode Island, Kinston, RI, 02881, USA

    Bingfang Yan

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  1. Yuxin Li
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  2. Qiang Shen
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  3. Hee-Tae Kim
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  4. Reid P. Bissonnette
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  5. William W. Lamph
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  7. Powel H. Brown
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Correspondence to Powel H. Brown.

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Li, Y., Shen, Q., Kim, HT. et al. The rexinoid bexarotene represses cyclin D1 transcription by inducing the DEC2 transcriptional repressor. Breast Cancer Res Treat 128, 667–677 (2011). https://doi.org/10.1007/s10549-010-1083-9

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  • DOI: https://doi.org/10.1007/s10549-010-1083-9

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