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Estrogen receptor α is the major driving factor for growth in tamoxifen-resistant breast cancer and supported by HER/ERK signaling

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Abstract

Resistance to tamoxifen is a major clinical challenge in the treatment of breast cancer; however, it is still unclear which signaling pathways are the major drivers of tamoxifen-resistant growth. To characterize resistance mechanisms, we have generated different tamoxifen-resistant breast cancer cell lines from MCF-7. In this model, we investigated whether signaling from human epidermal growth factor receptors (HERs), their downstream kinases, or from the estrogen receptor α (ERα) was driving tamoxifen-resistant cell growth. Increased expression of EGFR and increased phosphorylation of HER3 were observed upon acquisition of tamoxifen resistance, and the extracellular activated kinase (ERK) signaling pathway was highly activated in the resistant cells. The EGFR inhibitor gefitinib and the ERK pathway inhibitor U0126 resulted in partial and preferential growth inhibition of tamoxifen-resistant cells. All the tamoxifen-resistant cell lines retained ERα expression but at a lower level compared to that in MCF-7. Importantly, we showed via ERα knockdown that the tamoxifen-resistant cells were dependent on functional ERα for growth and we observed a clear growth stimulation of resistant cell lines with clinically relevant concentrations of tamoxifen and 4-OH-tamoxifen, indicating that tamoxifen-resistant cells utilize agonistic ERα stimulation by tamoxifen for growth. The tamoxifen-resistant cells displayed high phosphorylation of ERα at Ser118 in the presence of tamoxifen; however, treatment with U0126 neither affected the level of Ser118 phosphorylation nor expression of the ERα target Bcl-2, suggesting that ERK contributes to cell growth independently of ERα in our cell model. In support of this, combined treatment against ERα and ERK signaling in resistant cells was superior to single-agent treatment and as effective as fulvestrant treatment of MCF-7 cells. Together, these findings demonstrate that ERα is a major driver of growth in tamoxifen-resistant cells supported by HER/ERK growth signaling, implying that combined targeting of these pathways may have a clinical potential for overcoming tamoxifen resistance.

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Acknowledgments

We thank Jane Lind Christensen for excellent technical assistance. This study was supported by grants from the Danish Medical Research Council (271-07-0704), Danish Cancer Society (R38-A1682), A Race Against Breast Cancer, and Balslev Foundation.

Conflict of interest

  The authors declare that they have no conflict of interests.

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Authors and Affiliations

  1. Breast Cancer Group, Unit of Cell Death and Metabolism, Danish Cancer Society Research Center, Strandboulevarden 49, 2100, Copenhagen Ø, Denmark

    Susan Thrane, Anne E. Lykkesfeldt, Mathilde S. Larsen & Christina W. Yde

  2. Department of Clinical Biochemistry, Aarhus University Hospital, Nørrebrogade 44, 8000, Aarhus C, Denmark

    Boe S. Sorensen

Authors
  1. Susan Thrane
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  2. Anne E. Lykkesfeldt
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  3. Mathilde S. Larsen
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  4. Boe S. Sorensen
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  5. Christina W. Yde
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Correspondence to Christina W. Yde.

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Thrane, S., Lykkesfeldt, A.E., Larsen, M.S. et al. Estrogen receptor α is the major driving factor for growth in tamoxifen-resistant breast cancer and supported by HER/ERK signaling. Breast Cancer Res Treat 139, 71–80 (2013). https://doi.org/10.1007/s10549-013-2485-2

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  • DOI: https://doi.org/10.1007/s10549-013-2485-2

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