Department of Immunology, Genetics and Pathology, Uppsala University, Science for Life Laboratory, 751 85 Uppsala, Sweden.
Department of Chemistry, Uppsala University, Science for Life Laboratory, 75123 Uppsala, Sweden.
Nat Commun. 2017 Jun 15;8:15840. doi: 10.1038/ncomms15840.
The discovery of oestrogen receptor β (ERβ/ESR2) was a landmark discovery. Its reported expression and homology with breast cancer pharmacological target ERα (ESR1) raised hopes for improved endocrine therapies. After 20 years of intense research, this has not materialized. We here perform a rigorous validation of 13 anti-ERβ antibodies, using well-characterized controls and a panel of validation methods. We conclude that only one antibody, the rarely used monoclonal PPZ0506, specifically targets ERβ in immunohistochemistry. Applying this antibody for protein expression profiling in 44 normal and 21 malignant human tissues, we detect ERβ protein in testis, ovary, lymphoid cells, granulosa cell tumours, and a subset of malignant melanoma and thyroid cancers. We do not find evidence of expression in normal or cancerous human breast. This expression pattern aligns well with RNA-seq data, but contradicts a multitude of studies. Our study highlights how inadequately validated antibodies can lead an exciting field astray.
雌激素受体 β（ERβ/ESR2）的发现是一个具有里程碑意义的发现。据报道，它与乳腺癌药物靶点 ERα（ESR1）的表达和同源性，燃起了人们对改善内分泌治疗的希望。经过 20 年的深入研究，这一愿望并未实现。我们在这里使用经过充分验证的对照和一系列验证方法，对 13 种抗 ERβ 抗体进行了严格的验证。我们得出的结论是，只有一种抗体，即很少使用的单克隆抗体 PPZ0506，在免疫组织化学中特异性地针对 ERβ。我们应用这种抗体对 44 份正常和 21 份恶性人类组织的蛋白质表达谱进行分析，在睾丸、卵巢、淋巴样细胞、颗粒细胞瘤以及一部分恶性黑色素瘤和甲状腺癌中检测到 ERβ 蛋白。我们没有发现 ERβ 在正常或癌变的人类乳腺中的表达证据。这种表达模式与 RNA-seq 数据非常吻合，但与大量研究相矛盾。我们的研究强调了未经充分验证的抗体如何误导一个令人兴奋的领域。
