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Progestin suppression of miR-29 potentiates dedifferentiation of breast cancer cells via KLF4

Oncogene volume 32, pages 2555–2564 (2013)Cite this article

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Abstract

The female hormone progesterone (P4) promotes the expansion of stem-like cancer cells in estrogen receptor (ER)- and progesterone receptor (PR)-positive breast tumors. The expanded tumor cells lose expression of ER and PR, express the tumor-initiating marker CD44, the progenitor marker cytokeratin 5 (CK5) and are more resistant to standard endocrine and chemotherapies. The mechanisms underlying this hormone-stimulated reprogramming have remained largely unknown. In the present study, we investigated the role of microRNAs in progestin-mediated expansion of this dedifferentiated tumor cell population. We demonstrate that P4 rapidly downregulates miR-29 family members, particularly in the CD44+ cell population. Downregulation of miR-29 members potentiates the expansion of CK5+ and CD44+ cells in response to progestins, and results in increased stem-like properties in vitro and in vivo. We demonstrate that miR-29 directly targets Krüppel-like factor 4 (KLF4), a transcription factor required for the reprogramming of differentiated cells to pluripotent stem cells, and for the maintenance of breast cancer stem cells. These results reveal a novel mechanism, whereby progestins increase the stem cell-like population in hormone-responsive breast cancers, by decreasing miR-29 to augment PR-mediated upregulation of KLF4. Elucidating the mechanisms whereby hormones mediate the expansion of stem-like cells furthers our understanding of the progression of hormone-responsive breast cancers.

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Acknowledgements

We thank the University of Colorado Cancer Center Cytometry and Cell Sorting Shared Resource Facility supported by P30CA046934. The human CK5 promoter (KRT5) was a gift from Elaine Fuchs (The Rockefeller University). NIH F31CA165668-01 supported ENH. NIH RO1CA140985 supported CAS. DOD BCRP Postdoctoral Fellowship W81XWH-11-1-0101 (DMC) and DOD Idea Award BCRP W81XWH-11-1-0210 (CAS, JKR) supported this work.

DMC performed most of the studies. JFS, PH created BT474 stable cell lines and performed reverse-transcriptase PCRs in BT474 cells. ENH quantified immunofluorescence images. SDA created the KRT5-luciferase reporter. NSS performed IHC. DMC, JFS, CAS, BMJ and JKR contributed intellectual input towards the design, implementation and interpretation of results. DMC wrote the manuscript. BMJ, CAS, JKR 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 Denver, Aurora, CO, USA

    D M Cittelly, J Finlay-Schultz, E N Howe, N S Spoelstra, S D Axlund, P Hendricks, C A Sartorius & J K Richer

  2. Department of Medicine, Division of Endocrinology, University of Colorado Denver, Aurora, CO, USA

    B M Jacobsen

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  1. D M Cittelly
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Cittelly, D., Finlay-Schultz, J., Howe, E. et al. Progestin suppression of miR-29 potentiates dedifferentiation of breast cancer cells via KLF4. Oncogene 32, 2555–2564 (2013). https://doi.org/10.1038/onc.2012.275

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