Estrogen receptor beta increases sensitivity to enzalutamide in androgen receptor-positive triple-negative breast cancer

Abstract

Purpose

Androgen receptor (AR) is playing an important role in the progression of a subset of TNBC. We evaluated the impact of ERβ expression along with anti-AR drugs in AR-positive TNBC.

Methods

ERβ expression was examined in AR-positive TNBC cell line using MTT assay, scratch and Annexin V-FITC assay in the presence or absence of anti-androgens. Protein levels of involved molecules were assessed using Western blot. Receptors’ localization was detected by immunofluorescence and their physical association was examined using proximity ligation assay (PLA), which enables the visualization of interacting proteins in fixed cells and tissues.

Results

Transient transfection of ERβ in MDA-MB 453 AR-positive TNBC cell line significantly inhibited cell proliferation, metastatic potential and induced apoptosis. ERβ expression reversed the aggravating role of AR in both indirect and direct ways. Indirectly, ERβ decreased AR activation through the inhibition of PI3K/AKT signaling pathway. Directly, ERβ formed heterodimers with AR in MDA-MB 453 cells and in human tissue samples impeding AR from forming homodimers. Enzalutamide is a more potent anti-androgen in AR + TNBC compared to bicalutamide. ERβ expression increased the sensitivity of MDA-MB 453 cells to anti-androgens and especially to enzalutamide. The administration of enzalutamide enhanced AR:ERβ heterodimers formation increasing the anti-tumor capacity of ERβ.

Conclusions

Collectively, our results provide evidence for a novel mechanism by which ERβ exerts oncosuppressive effect in AR-positive TBNC through direct and indirect interactions with AR. Moreover, ERβ expression may identify a new subset of TNBC that would respond more favorable to anti-androgens.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

Sponsored by ASTELLAS PHARMA EUROPE LTD ISR GR-72-RG-35.

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Authors

Contributions

AA, PS, ST, IZ, DT, AK, AK, DT, EX, and MK made substantial contributions to acquisition, analysis, and interpretation of data. AA, AGP, and MVK made substantial contributions in the conception, design, and interpretation of the data. AA, AGP, and MVK made substantial contributions in drafting the manuscript and revising it critically for important intellectual content.

Corresponding authors

Correspondence to Athanasios G. Papavassiliou or Michalis V. Karamouzis.

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The authors declare that they have no competing interests.

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The protocol of this study was approved by the National and Kapodistrian University Ethics Committee following the principles of the Declaration of Helsinki.

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Anestis, A., Sarantis, P., Theocharis, S. et al. Estrogen receptor beta increases sensitivity to enzalutamide in androgen receptor-positive triple-negative breast cancer. J Cancer Res Clin Oncol 145, 1221–1233 (2019). https://doi.org/10.1007/s00432-019-02872-9

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Keywords

  • Triple-negative breast cancer
  • Androgen receptor
  • Estrogen receptor β (ERβ)
  • Enzalutamide
  • Bicalutamide