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Cross talk between progesterone receptors and retinoic acid receptors in regulation of cytokeratin 5-positive breast cancer cells

Oncogene volume 36, pages 6074–6084 (2017)Cite this article

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Abstract

Half of estrogen receptor-positive breast cancers contain a subpopulation of cytokeratin 5 (CK5)-expressing cells that are therapy resistant and exhibit increased cancer stem cell (CSC) properties. We and others have demonstrated that progesterone (P4) increases CK5+ breast cancer cells. We previously discovered that retinoids block P4 induction of CK5+ cells. Here we investigated the mechanisms by which progesterone receptors (PR) and retinoic acid receptors (RAR) regulate CK5 expression and breast CSC activity. After P4 treatment, sorted CK5+ compared to CK5− cells were more tumorigenic in vivo. In vitro, P4-treated breast cancer cells formed larger mammospheres and silencing of CK5 using small hairpin RNA abolished this P4-dependent increase in mammosphere size. Retinoic acid (RA) treatment blocked the P4 increase in CK5+ cells and prevented the P4 increase in mammosphere size. Dual small interfering RNA (siRNA) silencing of RARα and RARγ reversed RA blockade of P4-induced CK5. Using promoter deletion analysis, we identified a region 1.1 kb upstream of the CK5 transcriptional start site that is necessary for P4 activation and contains a putative progesterone response element (PRE). We confirmed by chromatin immunoprecipitation that P4 recruits PR to the CK5 promoter near the −1.1 kb essential PRE, and also to a proximal region near −130 bp that contains PRE half-sites and a RA response element (RARE). RA induced loss of PR binding only at the proximal site. Interestingly, RARα was recruited to the −1.1 kb PRE and the −130 bp PRE/RARE regions with P4, but not RA alone or RA plus P4. Treatment of breast cancer xenografts in vivo with the retinoid fenretinide reduced the accumulation of CK5+ cells during estrogen depletion. This reduction, together with the inhibition of CK5+ cell expansion through RAR/PR cross talk, may explain the efficacy of retinoids in prevention of some breast cancer recurrences.

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Acknowledgements

We thank the University of Colorado Cancer Center Flow Cytometry, Biorepository Core, and Tissue Culture Cores supported by P30CA046934 and the University of Colorado Department of Pathology Sequencing Core for their technical assistance and services. We thank Andrea Osypuk and Story Wilson for their assistance with Aperio imaging and analysis. This work was supported by grants from the Colorado Clinical and Translational Sciences Institute NIH TL1 TR001081 (LMF), National Institutes of Health grants NIH F31 CA210519 (LMF), NIH 2R01 CA140985 (CAS) and Breast Cancer Research Foundation (16-072, CAS, co-PI).

Author contributions

LMF performed most of the studies. OM performed experiments in Figure 2a and b, and Figure 5b and c. JF-S performed experiments in Table 1 and Supplementary Figure 1. SKN engineered and provided reagents for Figure 4. LMF, JF-S, DVL, SKN and CAS contributed intellectual design and interpretation of results. LMF wrote the manuscript. JF-S, SKN and CAS provided editorial assistance. All authors read and approved the final manuscript.

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Authors and Affiliations

  1. Department of Pathology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    L M Fettig, O McGinn, J Finlay-Schultz, S K Nordeen & C A Sartorius

  2. Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, CO, USA

    D V LaBarbera

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  1. L M Fettig
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Correspondence to C A Sartorius.

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Fettig, L., McGinn, O., Finlay-Schultz, J. et al. Cross talk between progesterone receptors and retinoic acid receptors in regulation of cytokeratin 5-positive breast cancer cells. Oncogene 36, 6074–6084 (2017). https://doi.org/10.1038/onc.2017.204

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