Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity.
Comprehensive assessment of estrogen receptor beta antibodies in cancer cell line models and tissue reveals critical limitations in reagent specificity.
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DOI：
10.1016/j.mce.2016.11.016
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发表时间：
2017-01-15
影响因子：
4.1
通讯作者：
Carroll JS
中科院分区：
文献类型：
--
作者：
Nelson AW;Groen AJ;Miller JL;Warren AY;Holmes KA;Tarulli GA;Tilley WD;Katzenellenbogen BS;Hawse JR;Gnanapragasam VJ;Carroll JS
雌激素受体-β（ER β）与许多癌症有关。在前列腺癌和乳腺癌中，它的功能是有争议的，但遗传学研究暗示了它在癌症进展中的作用。围绕ER β的大部分混乱源于未经充分验证的抗体，但已成为破译其作用的标准工具。使用ER β诱导细胞系统，我们评估了常用的ER β抗体，并显示最常用的抗体之一NCL-ER-BETA对ER β是非特异性的。其他抗体具有有限的ER β特异性或仅在一种实验模式中具有特异性。ER β通常在MCF-7（乳腺癌）和LNCaP（前列腺癌）癌细胞系中进行研究，但我们使用经验证的抗体和独立的基于质谱的方法，在这两种细胞系中均未发现ER β表达。我们的发现质疑了使用NCL-ER-BETA抗体或LNCaP和MCF-7细胞系得出的关于ER β的结论。我们描述了稳健的试剂，可在多种实验方法和临床样本中检测ER β。ER β在前列腺癌和乳腺癌中很重要，但其作用存在争议。ER β抗体存在问题，具有不同的特异性。我们测试了一组ER β抗体，并显示最常用的是非特异性的。在多种实验方法中验证了两种抗体。使用多种技术，我们显示用于研究ER β的细胞系缺乏其表达。
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. 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.