Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Control of mammary stem cell function by steroid hormone signalling


The ovarian hormones oestrogen and progesterone profoundly influence breast cancer risk1,2,3, underpinning the benefit of endocrine therapies in the treatment of breast cancer4. Modulation of their effects through ovarian ablation or chemoprevention strategies also significantly decreases breast cancer incidence5,6. Conversely, there is an increased risk of breast cancer associated with pregnancy in the short term7. The cellular mechanisms underlying these observations, however, are poorly defined. Here we demonstrate that mouse mammary stem cells (MaSCs)8,9 are highly responsive to steroid hormone signalling, despite lacking the oestrogen and progesterone receptors10. Ovariectomy markedly diminished MaSC number and outgrowth potential in vivo, whereas MaSC activity increased in mice treated with oestrogen plus progesterone. Notably, even three weeks of treatment with the aromatase inhibitor letrozole was sufficient to reduce the MaSC pool. In contrast, pregnancy led to a transient 11-fold increase in MaSC numbers, probably mediated through paracrine signalling from RANK ligand. The augmented MaSC pool indicates a cellular basis for the short-term increase in breast cancer incidence that accompanies pregnancy. These findings further indicate that breast cancer chemoprevention may be achieved, in part, through suppression of MaSC function.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Steroid hormone deprivation reduces MaSC activity.
Figure 2: Marked increase in the number of MaSCs in mid-pregnancy.
Figure 3: RANKL is involved in MaSC activation observed during pregnancy.

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

All microarray data are available from the Gene Expression Omnibus database ( under accession codes GSE20401 and GSE20402.


  1. Clemons, M. & Goss, P. Estrogen and the risk of breast cancer. N. Engl. J. Med. 344, 276–285 (2001)

    CAS  Article  Google Scholar 

  2. Hankinson, S. E., Colditz, G. A. & Willett, W. C. Towards an integrated model for breast cancer etiology: the lifelong interplay of genes, lifestyle, and hormones. Breast Cancer Res. 6, 213–218 (2004)

    CAS  Article  Google Scholar 

  3. Pike, M. C., Spicer, D. V., Dahmoush, L. & Press, M. F. Estrogens, progestogens, normal breast cell proliferation, and breast cancer risk. Epidemiol. Rev. 15, 17–35 (1993)

    CAS  Article  Google Scholar 

  4. Early Breast Cancer Trialists’ Collaborative Group Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet 365, 1687–1717 (2005)

    Article  Google Scholar 

  5. Parker, W. H. et al. Ovarian conservation at the time of hysterectomy and long-term health outcomes in the nurses’ health study. Obstet. Gynecol. 113, 1027–1037 (2009)

    Article  Google Scholar 

  6. Visvanathan, K. et al. American society of clinical oncology clinical practice guideline update on the use of pharmacologic interventions including tamoxifen, raloxifene, and aromatase inhibition for breast cancer risk reduction. J. Clin. Oncol. 27, 3235–3258 (2009)

    CAS  Article  Google Scholar 

  7. Lambe, M. et al. Transient increase in the risk of breast cancer after giving birth. N. Engl. J. Med. 331, 5–9 (1994)

    CAS  Article  Google Scholar 

  8. Shackleton, M. et al. Generation of a functional mammary gland from a single stem cell. Nature 439, 84–88 (2006)

    ADS  CAS  Article  Google Scholar 

  9. Stingl, J. et al. Purification and unique properties of mammary epithelial stem cells. Nature 439, 993–997 (2006)

    ADS  CAS  Article  Google Scholar 

  10. Asselin-Labat, M. L. et al. Steroid hormone receptor status of mouse mammary stem cells. J. Natl Cancer Inst. 98, 1011–1014 (2006)

    CAS  Article  Google Scholar 

  11. Anderson, E. & Clarke, R. B. Steroid receptors and cell cycle in normal mammary epithelium. J. Mammary Gland Biol. Neoplasia 9, 3–13 (2004)

    Article  Google Scholar 

  12. Mueller, S. O., Clark, J. A., Myers, P. H. & Korach, K. S. Mammary gland development in adult mice requires epithelial and stromal estrogen receptor alpha. Endocrinology 143, 2357–2365 (2002)

    CAS  Article  Google Scholar 

  13. Mallepell, S., Krust, A., Chambon, P. & Brisken, C. Paracrine signaling through the epithelial estrogen receptor alpha is required for proliferation and morphogenesis in the mammary gland. Proc. Natl Acad. Sci. USA 103, 2196–2201 (2006)

    ADS  CAS  Article  Google Scholar 

  14. Brisken, C. et al. A paracrine role for the epithelial progesterone receptor in mammary gland development. Proc. Natl Acad. Sci. USA 95, 5076–5081 (1998)

    ADS  CAS  Article  Google Scholar 

  15. Lydon, J. P. et al. Mice lacking progesterone receptor exhibit pleiotropic reproductive abnormalities. Genes Dev. 9, 2266–2278 (1995)

    CAS  Article  Google Scholar 

  16. Mulac-Jericevic, B., Lydon, J. P., DeMayo, F. J. & Conneely, O. M. Defective mammary gland morphogenesis in mice lacking the progesterone receptor B isoform. Proc. Natl Acad. Sci. USA 100, 9744–9749 (2003)

    ADS  CAS  Article  Google Scholar 

  17. Forbes, J. F. et al. Effect of anastrozole and tamoxifen as adjuvant treatment for early-stage breast cancer: 100-month analysis of the ATAC trial. Lancet Oncol. 9, 45–53 (2008)

    Article  Google Scholar 

  18. Visvader, J. E. Keeping abreast of the mammary epithelial hierarchy and breast tumorigenesis. Genes Dev. 23, 2563–2577 (2009)

    CAS  Article  Google Scholar 

  19. Lim, E. et al. Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nature Med. 15, 907–913 (2009)

    CAS  Article  Google Scholar 

  20. Fernandez-Valdivia, R. et al. Transcriptional response of the murine mammary gland to acute progesterone exposure. Endocrinology 149, 6236–6250 (2008)

    CAS  Article  Google Scholar 

  21. Ciarloni, L., Mallepell, S. & Brisken, C. Amphiregulin is an essential mediator of estrogen receptor alpha function in mammary gland development. Proc. Natl Acad. Sci. USA 104, 5455–5460 (2007)

    ADS  CAS  Article  Google Scholar 

  22. Asselin-Labat, M. L. et al. Gata-3 is an essential regulator of mammary-gland morphogenesis and luminal-cell differentiation. Nature Cell Biol. 9, 201–209 (2007)

    CAS  Article  Google Scholar 

  23. Fisher, C. R., Graves, K. H., Parlow, A. F. & Simpson, E. R. Characterization of mice deficient in aromatase (ArKO) because of targeted disruption of the cyp19 gene. Proc. Natl Acad. Sci. USA 95, 6965–6970 (1998)

    ADS  CAS  Article  Google Scholar 

  24. Lydon, J. P., Sivaraman, L. & Conneely, O. M. A reappraisal of progesterone action in the mammary gland. J. Mammary Gland Biol. Neoplasia 5, 325–338 (2000)

    CAS  Article  Google Scholar 

  25. Srivastava, S. et al. Receptor activator of NF-κB ligand induction via Jak2 and Stat5a in mammary epithelial cells. J. Biol. Chem. 278, 46171–46178 (2003)

    CAS  Article  Google Scholar 

  26. Fata, J. E. et al. The osteoclast differentiation factor osteoprotegerin-ligand is essential for mammary gland development. Cell 103, 41–50 (2000)

    CAS  Article  Google Scholar 

  27. Fernandez-Valdivia, R. et al. The RANKL signaling axis is sufficient to elicit ductal side-branching and alveologenesis in the mammary gland of the virgin mouse. Dev. Biol. 328, 127–139 (2009)

    CAS  Article  Google Scholar 

  28. Gonzalez-Suarez, E. et al. RANK overexpression in transgenic mice with mouse mammary tumor virus promoter-controlled RANK increases proliferation and impairs alveolar differentiation in the mammary epithelia and disrupts lumen formation in cultured epithelial acini. Mol. Cell. Biol. 27, 1442–1454 (2007)

    CAS  Article  Google Scholar 

  29. Kim, N. S. et al. Receptor activator of NF-κB ligand regulates the proliferation of mammary epithelial cells via Id2. Mol. Cell. Biol. 26, 1002–1013 (2006)

    CAS  Article  Google Scholar 

  30. Howell, A. The endocrine prevention of breast cancer. Best Pract. Res. Clin. Endocrinol. Metab. 22, 615–623 (2008)

    CAS  Article  Google Scholar 

  31. Hu, Y. & Smyth, G. K. ELDA: Extreme limiting dilution analysis for comparing depleted and enriched populations in stem cell and other assays. J. Immunol. Methods 347, 70–78 (2009)

    CAS  Article  Google Scholar 

  32. Dontu, G. et al. In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev. 17, 1253–1270 (2003)

    CAS  Article  Google Scholar 

  33. Gentleman, R. C. et al. Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80 (2004)

    Article  Google Scholar 

  34. Ritchie, M. E. et al. A comparison of background correction methods for two-colour microarrays. Bioinformatics 23, 2700–2707 (2007)

    CAS  Article  Google Scholar 

  35. Smyth, G. K. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3, 1–28 (2004)

    ADS  MathSciNet  Article  Google Scholar 

  36. Ritchie, M. E. et al. Empirical array quality weights in the analysis of microarray data. BMC Bioinform. 7, 261 (2006)

    Article  Google Scholar 

  37. Smyth, G. K., Michaud, J. & Scott, H. S. Use of within-array replicate spots for assessing differential expression in microarray experiments. Bioinformatics 21, 2067–2075 (2005)

    CAS  Article  Google Scholar 

Download references


We are grateful to A. Morcom and T. Ward for technical assistance, T. Bouras for advice, S. Mihajlovic for histology and F. Battye for FACS support. We thank C. Clarke for providing the hPRa7 antibody, A. Burgess for AG1478, A. Parlow for prolactin, and the Australian Genome Research Facility for RNA bioanalyses. M.-L.A.-L. is supported by an Australian Research Council Postdoctoral Fellowship. This work was supported by the Victorian Breast Cancer Research Consortium (J.E.V. and G.J.L.), the National Health and Medical Research Council (Australia), the Susan G. Komen Foundation, the National Breast Cancer Foundation and the Australian Cancer Research Foundation.

Author information

Authors and Affiliations



M.-L.A.-L. conceptualized and designed the experiments and performed most of the experiments and data analysis; F.V. performed transplantation experiments and analysis; J.S. performed in vitro inhibitor experiments; B.P. performed quantitative RT–PCR; D.W. and G.K.S. performed bioinformatics analyses; E.R.S. provided ArKO mice and advice; H.Y. provided anti-RANKL inhibitor and advice; T.J.M. provided advice and helped design RANKL experiments; J.E.V. and G.J.L. conceived and directed the study, and J.E.V., G.J.L. and M.-L.A.-L. wrote the manuscript.

Corresponding authors

Correspondence to Geoffrey J. Lindeman or Jane E. Visvader.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures S1-S7 with legends. The Supplementary Tables were added on 14 April 2010. A small correction was made to Supplementary Table 5 on 19 May 2010. (PDF 2300 kb)

Supplementary Table 1

This file contains the gene profiling data (Log2-intensity and the log2-fold-change) for mammary populations from control versus ovariectomized (Ovx) and 12.5 day pregnant mice. (XLS 623 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Asselin-Labat, ML., Vaillant, F., Sheridan, J. et al. Control of mammary stem cell function by steroid hormone signalling. Nature 465, 798–802 (2010).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:

Further reading


By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing