Huang Ruili, Sakamuru Srilatha, Martin Matt T, Reif David M, Judson Richard S, Houck Keith A, Casey Warren, Hsieh Jui-Hua, Shockley Keith R, Ceger Patricia, Fostel Jennifer, Witt Kristine L, Tong Weida, Rotroff Daniel M, Zhao Tongan, Shinn Paul, Simeonov Anton, Dix David J, Austin Christopher P, Kavlock Robert J, Tice Raymond R, Xia Menghang
NIH Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD 20850, USA.
National Center for Computational Toxicology, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711, USA.
Sci Rep. 2014 Jul 11;4:5664. doi: 10.1038/srep05664.
The U.S. Tox21 program has screened a library of approximately 10,000 (10K) environmental chemicals and drugs in three independent runs for estrogen receptor alpha (ERα) agonist and antagonist activity using two types of ER reporter gene cell lines, one with an endogenous full length ERα (ER-luc; BG1 cell line) and the other with a transfected partial receptor consisting of the ligand binding domain (ER-bla; ERα β-lactamase cell line), in a quantitative high-throughput screening (qHTS) format. The ability of the two assays to correctly identify ERα agonists and antagonists was evaluated using a set of 39 reference compounds with known ERα activity. Although both assays demonstrated adequate (i.e. >80%) predictivity, the ER-luc assay was more sensitive and the ER-bla assay more specific. The qHTS assay results were compared with results from previously published ERα binding assay data and showed >80% consistency. Actives identified from both the ER-bla and ER-luc assays were analyzed for structure-activity relationships (SARs) revealing known and potentially novel ERα active structure classes. The results demonstrate the feasibility of qHTS to identify environmental chemicals with the potential to interact with the ERα signaling pathway and the two different assay formats improve the confidence in correctly identifying these chemicals.
美国毒物21计划使用两种雌激素受体α（ERα）报告基因细胞系，以定量高通量筛选（qHTS）形式，对约10000种（10K）环境化学品和药物的文库进行了三次独立检测，以检测其ERα激动剂和拮抗剂活性。一种细胞系内源性全长ERα（ER-luc；BG1细胞系），另一种细胞系转染了由配体结合域组成的部分受体（ER-bla；ERαβ-内酰胺酶细胞系）。使用一组39种具有已知ERα活性的参考化合物评估了这两种检测方法正确识别ERα激动剂和拮抗剂的能力。虽然两种检测方法都显示出足够的（即>80%）预测性，但ER-luc检测更敏感，ER-bla检测更具特异性。将qHTS检测结果与先前发表的ERα结合检测数据结果进行比较，一致性>80%。对从ER-bla和ER-luc检测中鉴定出的活性物质进行了构效关系（SAR）分析，揭示了已知的和潜在的新型ERα活性结构类别。结果证明了qHTS用于识别可能与ERα信号通路相互作用的环境化学品的可行性，并且两种不同的检测形式提高了正确识别这些化学品的可信度。
