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
Cancer Research UK Cambridge Institute, University of Cambridge, Robinson Way, Cambridge, CB2 ORE, UK; Academic Urology Group, Department of Surgery, University of Cambridge, Cambridge, CB2 0QQ, UK; Department of Urology, Addenbrooke's Hospital, Cambridge University Hospitals NHS Foundation Trust, Hills Road, Cambridge, CB2 0QQ, UK.
Cancer Research UK Cambridge Institute, University of Cambridge, Robinson Way, Cambridge, CB2 ORE, UK.
Mol Cell Endocrinol. 2017 Jan 15;440:138-150. doi: 10.1016/j.mce.2016.11.016. Epub 2016 Nov 23.
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.
雌激素受体-β（ERβ）与多种癌症有关。在前列腺癌和乳腺癌中，其功能存在争议，但遗传学研究表明它在癌症进展中起作用。围绕ERβ的许多困惑源于未充分验证的抗体，但这些抗体已成为解读其作用的标准工具。我们使用ERβ诱导细胞系统评估了常用的ERβ抗体，结果表明最常用的抗体之一NCL-ER-BETA对ERβ不具有特异性。其他抗体的ERβ特异性有限，或仅在一种实验模式中具有特异性。ERβ通常在MCF-7（乳腺癌）和LNCaP（前列腺癌）癌细胞系中进行研究，但我们使用经过验证的抗体和基于质谱的独立方法，在这两种细胞系中均未发现ERβ表达。我们的研究结果对使用NCL-ER-BETA抗体或LNCaP和MCF-7细胞系得出的关于ERβ的结论提出了质疑。我们描述了在多种实验方法和临床样本中均能检测到ERβ的可靠试剂。
