Abstract
Progesterone (P4) has emerged as an important hormone-regulating mammary stem cell (MaSC) populations. In breast cancer, P4 and synthetic analogs increase the number of stem-like cells within luminal estrogen receptor (ER)- and progesterone receptor (PR)-positive breast cancers. These cells gain expression of de-differentiated cell markers CD44 and cytokeratin 5 (CK5), lose luminal markers ER and PR, and are more therapy resistant. We previously described that P4 downregulation of microRNA (miR)-29a contributes to the expansion of CD44high and CK5+ cells. Here we investigated P4 downregulation of miR-141, a member of the miR-200 family of tumor suppressors, in facilitating an increase in stem-like breast cancer cells. miR-141 was the sole member of the miR-200 family P4-downregulated at the mature miRNA level in luminal breast cancer cell lines. Stable inhibition of miR-141 alone increased the CD44high population, and potentiated P4-mediated increases in both CD44high and CK5+ cells. Loss of miR-141 enhanced both mammosphere formation and tumor initiation. miR-141 directly targeted both PR and signal transducer and activator of transcription 5A (Stat5a), transcription factors important for MaSC expansion. miR-141 depletion increased PR protein levels, even in cell lines where PR expression is estrogen dependent. Stat5a suppression via small interfering RNA or a small-molecule inhibitor reduced the P4-dependent increase in CK5+ and CD44high cells. These data support a mechanism by which P4-triggered loss of miR-141 facilitates breast cancer cell de-differentiation through deregulation of PR and Stat5a, two transcription factors important for controlling mammary cell fate.
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Acknowledgements
We thank the University of Colorado Cancer Center Flow Cytometry and Tissue Culture Cores supported by P30CA046934 and the University of Colorado, Department of Pathology Sequencing Core for their technical assistance and services. This work was supported by Department of Defense BCRP grants W81XWH-11-1-0210 (CAS, JKR) and W81XWH-11-1-0101 (DMC) and NIH R01 CA140985 (CAS). PK was supported by NIH K08 CA164048.
Author Contributions
JF-S and PH performed most of the studies. DMC performed experiments in Figures 2a, b and 5c, d. PP performed experiments in Figure 3a. PK provided the PR exonic-binding site for miR-141 (Figure 4c) and technical advice. JF-S, DMC, BMJ, JKR and CAS contributed intellectual design and interpretation of results. JFS wrote the manuscript. BMJ, JKR and CAS provided editorial assistance. All authors read and approved the final manuscript.
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Finlay-Schultz, J., Cittelly, D., Hendricks, P. et al. Progesterone downregulation of miR-141 contributes to expansion of stem-like breast cancer cells through maintenance of progesterone receptor and Stat5a. Oncogene 34, 3676–3687 (2015). https://doi.org/10.1038/onc.2014.298
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DOI: https://doi.org/10.1038/onc.2014.298
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