Breast Cancer Research and Treatment

, Volume 146, Issue 3, pp 515–523 | Cite as

RANKL expression in normal and malignant breast tissue responds to progesterone and is up-regulated during the luteal phase

  • Hong Hu
  • Jun Wang
  • Akash Gupta
  • Ali Shidfar
  • Daniel Branstetter
  • Oukseub Lee
  • David Ivancic
  • Megan Sullivan
  • Robert T. ChattertonJr.
  • William C. Dougall
  • Seema A. Khan
Preclinical study


The receptor activator of nuclear factor-κB ligand (RANKL) acts as a paracrine factor in progesterone-induced mammary epithelial proliferation and tumorigenesis. This evidence comes mainly from mouse models. Our aim was to examine whether RANKL expression in human normal and malignant breast is under the control of progesterone throughout the menstrual cycle. Breast epithelial samples were obtained by random fine needle aspiration (rFNA) of the contralateral unaffected breasts (CUB) of 18 breast cancer patients, with simultaneous serum hormone measurements. Genes correlated with serum progesterone levels were identified through Illumina microarray analysis. Validation was performed using qRT-PCR in rFNA samples from CUB of an additional 53 women and using immunohistochemistry in tissue microarrays of 61 breast cancer samples. Expression of RANKL, DIO2, and MYBPC1 was correlated with serum progesterone in CUB, and was significantly higher in luteal phase. RANKL and MYBPC1 mRNA expression were highly correlated between CUB and matched tumor samples. RANKL protein expression was also significantly increased in the luteal phase and highly correlated with serum progesterone levels in cancer samples, especially in hormone receptor positive tumors. The regulatory effects of progesterone on the expression of RANKL, DIO2, and MYBPC1 were confirmed in three-dimensional cultures of normal breast organoids. In normal breast and in breast cancer, RANKL mRNA and protein expression fluctuate with serum progesterone with highest levels in the luteal phase, suggesting that RANKL is a modulator of progesterone signaling in normal and malignant breast tissue and a potential biomarker of progesterone action and blockade.


RANKL Progesterone Menstrual cycle Luteal phase 



This study was funded by the Lynn Sage Cancer Research Foundation, R01CA12055501, and a generous donation from Ms. Dariel Eklund. Additional support was provided by the Amgen Inc.

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

10549_2014_3049_MOESM1_ESM.pdf (1.1 mb)
Supplementary material 1 (PDF 1178 kb)


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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Hong Hu
    • 1
  • Jun Wang
    • 1
  • Akash Gupta
    • 1
  • Ali Shidfar
    • 1
  • Daniel Branstetter
    • 5
  • Oukseub Lee
    • 1
  • David Ivancic
    • 1
  • Megan Sullivan
    • 2
  • Robert T. ChattertonJr.
    • 3
    • 4
  • William C. Dougall
    • 6
  • Seema A. Khan
    • 1
    • 4
  1. 1.Department of SurgeryNorthwestern University Feinberg School of MedicineChicagoUSA
  2. 2.Department of PathologyNorthwestern University Feinberg School of MedicineChicagoUSA
  3. 3.Department of Obstetrics & GynecologyNorthwestern University Feinberg School of MedicineChicagoUSA
  4. 4.Robert H. Lurie Comprehensive Cancer CenterNorthwestern University Feinberg School of MedicineChicagoUSA
  5. 5.Department of PathologyAmgen, IncSeattleUSA
  6. 6.Therapeutic Innovation UnitAmgen, IncSeattleUSA

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