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ERβ splice variant expression in four large cohorts of human breast cancer patient tumors

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Abstract

Though the role of Estrogen Receptor (ER)α in breast cancer has been studied extensively, there is little consensus about the role of alternative ER isoform ERβ in breast cancer biology. ERβ has significant sequence homology to ERα but is located on a different chromosome and maintains both overlapping and unique functional attributes. Five variants exist, resulting from alternative splicing of the C-terminal region of ERβ. The relevance of ERβ variants in breast cancer outcomes and response to therapy is difficult to assess because of conflicting reports in the literature, likely due to variable methods used to assess ERβ in patient tumors. Here, we quantitatively assess expression of ERβ splice variants on over 2,000 breast cancer patient samples. Antibodies against ERβ variants were validated for staining specificity in cell lines by siRNA knockdown of ESR2 and staining reproducibility on formalin-fixed paraffin-embedded tissue by quantitative immunofluorescence (QIF) using AQUA technology. We found antibodies against splice variants ERβ1 and ERβ5, but not ERβ2/cx, which were sensitive, specific, and reproducible. QIF staining of validated antibodies showed both ERβ1 and ERβ5 QIF scores, which have a normal (bell shaped) distribution on most cohorts assessed, and their expression is significantly associated with each other. Extensive survival analyses show that ERβ1 is not a prognostic or predictive biomarker for breast cancer. ERβ5 appears to be a context-dependent marker of worse outcome in HER2-positive and triple-negative patients, suggesting an unknown biological function in the absence of ERα.

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Acknowledgments

The authors of the study thank Pei Chao and Michael Stagner from Information Management Services (Silver Spring, MD) for data management support; the participants, physicians, pathologists, nurses, and interviewers from participating centers in Poland for their efforts during field-work; and Drs. Montserrat Garcia-Closas, Louise Brinton, Jolanta Lissowska, B. Peplonska for their contributions to the PBCS study design. Funding for this study was partially provided by the Breast Cancer Research Foundation.

Disclaimers

Dr. Rimm is a consultant for, and receives laboratory support from Genoptix Inc.

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Authors and Affiliations

  1. Department of Pathology, Yale University School of Medicine, BML116, 310 Cedar Street, PO Box 208023, New Haven, CT, 06520-8023, USA

    Hallie Wimberly & David L. Rimm

  2. Yale Center for Analytic Sciences, Yale University School of Public Health, New Haven, CT, USA

    Gang Han

  3. Department of Molecular Genetics and Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, M5S 1A8, Canada

    Dushanthi Pinnaduwage & Irene L. Andrulis

  4. Department of Biochemistry & Molecular Genetics, University of Manitoba, Winnipeg, MB, Canada

    Leigh C. Murphy

  5. Division of Cancer Epidemiology & Genetics, National Cancer Institute, Bethesda, MD, USA

    Xiaohong Rose Yang

  6. Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, USA

    Mark Sherman

  7. Division of Cancer Epidemiology & Genetics, Hormonal and Reproductive Epidemiology Branch, National Cancer Institute, Bethesda, MD, USA

    Jonine Figueroa

  8. Mount Sinai Hospital, Lunenfeld Tanenbaum Research Institute, Toronto, ON, M5G 1X5, Canada

    Dushanthi Pinnaduwage

Authors
  1. Hallie Wimberly
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  2. Gang Han
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  9. David L. Rimm
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Correspondence to David L. Rimm.

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Wimberly, H., Han, G., Pinnaduwage, D. et al. ERβ splice variant expression in four large cohorts of human breast cancer patient tumors. Breast Cancer Res Treat 146, 657–667 (2014). https://doi.org/10.1007/s10549-014-3050-3

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  • DOI: https://doi.org/10.1007/s10549-014-3050-3

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