Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity. (15th January 2017)

• Record Type: Journal Article
• Authors: Nelson, Adam W.; Groen, Arnoud J.; Miller, Jodi L.; Warren, Anne Y.; Holmes, Kelly A.; Tarulli, Gerard A.; Tilley, Wayne D.; Katzenellenbogen, Benita S.; Hawse, John R.; Gnanapragasam, Vincent J.; Carroll, Jason S.
• Is Part Of: Molecular and cellular endocrinology. Volume 440(2017)
• Page Start: 138
• Page End: 150
• Publication Date: 2017-01-15
• Subjects: Estrogen receptor beta -- Prostate -- Breast -- Cancer -- Antibody; Endocrinology -- Periodicals; Molecular biology -- Periodicals; Cytology -- Periodicals; Endocrinology -- Periodicals; Hormones -- Periodicals; Endocrinologie -- Périodiques; Cytology; Endocrinology; Molecular biology; Periodicals; 573.4
• DOI: 10.1016/j.mce.2016.11.016
• Languages: English
• ISSNs: 0303-7207

Abstract

Abstract: 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. Highlights: ERβ is important in prostate and breast cancer, but its role is controversial. ERβ antibodies are problematic, with varying specificity. We tested a panel of ERβ antibodies and show the most commonly used is non-specific. Two antibodies were validated across multiple experimental approaches. Using multiple techniques, we show cell lines used to study ERβ lack its expression.