Abstract
Purpose
Significant controversy exists regarding the expression patterns of estrogen receptor beta (ERβ) in normal and diseased breast tissue. To address this issue, we have validated two ERβ antibodies, optimized the IHC protocols for both antibodies and now report the expression patterns of ERβ in normal and malignant breast tissues.
Methods
ERβ antibody specificity was determined using western blot and IHC analysis. ERβ protein expression patterns were assessed via IHC in normal breast tissue and invasive breast carcinoma. Further, we report the detailed protocol of the ERβ IHC assay developed in our CAP/CLIA certified laboratory to provide a standardized method for future studies.
Results
We have confirmed the specificity of two independent ERβ monoclonal antibodies, one that detects total (i.e., full length plus splice variants 2–5, which do not include the ligand binding domain) ERβ protein (PPZ0506) and one that detects only the full-length form, which includes the ligand binding domain, of ERβ (PPG5/10). Using these two antibodies, we demonstrate that ERβ is highly expressed in normal human breast tissue as well as in 20–30% of invasive breast cancers. Further, these two antibodies exhibited similar staining patterns across multiple different tissues and were highly concordant with regard to determining ERβ positivity in breast cancers.
Conclusions
ERβ protein was shown to be abundant in the majority of normal breast epithelial cells and is present in 20–30% of breast cancers. Use of these two antibodies, along with their standardized IHC protocols, provide a reference for future studies aimed at determining the utility of ERβ as a prognostic and/or predictive biomarker in various tissues of benign or malignant states.
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Acknowledgements
We gratefully acknowledge the Mayo Clinic Immunostains Laboratory and the Mayo Clinic Pathology Research Core for their time and effort in developing the ERβ IHC assays.
Funding
This work was supported by the National Cancer Institute of the National Institutes of Health through the Mayo Clinic Breast Cancer SPORE: P50CA116201 (MPG and JRH), as well as the Eisenberg Foundation (JRH and MS) and Mayo Clinic.
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M.P.G. reports personal fees from Genomic Health, consulting fees from Lilly, Biovica, Novartis, Sermonix, Context Pharm, Pfizer, and Biotheranostics and grant funding from Pfizer and Lilly for efforts that are outside the context of the present study. All other authors declare that they have no conflicts of interest.
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This article does not contain any studies utilizing animals. All of the studies performed with the use of human tissue were in accordance with the ethical standards of the Institutional and/or National Research Committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Collection and assessment of ERβ in human tissues was approved by the Mayo Clinic IRB under protocols 16-007352, 13-000585 and 12-004582.
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Informed consent for tissue collection and use for future research was obtained from all individuals included in the study.
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Hawse, J.R., Carter, J.M., Aspros, K.G.M. et al. Optimized immunohistochemical detection of estrogen receptor beta using two validated monoclonal antibodies confirms its expression in normal and malignant breast tissues. Breast Cancer Res Treat 179, 241–249 (2020). https://doi.org/10.1007/s10549-019-05441-3
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DOI: https://doi.org/10.1007/s10549-019-05441-3