Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity
Molecular and Cellular Endocrinology
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Nelson, A., Groen, A., Miller, J., Warren, A., Holmes, K., Tarulli, G., Tilley, W., et al. (2017). Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity. Molecular and Cellular Endocrinology, 440 138-150. https://doi.org/10.1016/j.mce.2016.11.016
Estrogen Receptor-β (ERβ) has been implicated in many cancers. In prostate and breast cancer its function is controversial, but genetic studies implicate a role in cancer progression. Much of the confusion around ERβ stems from antibodies that are inadequately validated, yet have become standard tools for deciphering its role. Using an ERβ-inducible cell system we assessed commonly utilized ERβ antibodies and show that one of the most commonly used antibodies, NCL-ER-BETA, is non-specific for ERβ. Other antibodies have limited ERβ specificity or are only specific in one experimental modality. ERβ is commonly studied in MCF-7 (breast) and LNCaP (prostate) cancer cell lines, but we found no ERβ expression in either, using validated antibodies and independent mass spectrometry-based approaches. Our findings question conclusions made about ERβ using the NCL-ER-BETA antibody, or LNCaP and MCF-7 cell lines. We describe robust reagents, which detect ERβ across multiple experimental approaches and in clinical samples.
Antibody, Breast, Cancer, Estrogen receptor beta, Prostate
The authors acknowledge the Breast Cancer Research Foundation, Cancer Research UK, ERC Consolidator award (grant number 646876), Cambridge Biomedical Research Campus and Cambridge Cancer Centre, which fund the tissue bank and the Urology Biorepository.
Cancer Research UK (C14303_do not transfer)
ECH2020 EUROPEAN RESEARCH COUNCIL (ERC) (646876)
External DOI: https://doi.org/10.1016/j.mce.2016.11.016
This record's URL: https://www.repository.cam.ac.uk/handle/1810/263396
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: http://creativecommons.org/licenses/by-nc-nd/4.0/