Ceger Patricia, Allen David, Huang Ruili, Xia Menghang, Casey Warren
Integrated Laboratory Systems, Inc., Research Triangle Park, North Carolina, USA.
National Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland, USA.
ALTEX. 2015;32(4):287-96. doi: 10.14573/altex.1505121. Epub 2015 Jun 28.
In 2012, the BG1Luc4E2 estrogen receptor (ER) transactivation (TA) method (BG1Luc ER TA) was accepted by U.S. regulatory agencies and the Organisation for Economic Co-operation and Development to detect substances with ER agonist activity. The method is now part of the Tier 1 testing battery in the Environmental Protection Agency's Endocrine Disruptor Screening Program. The BG1Luc ER TA method uses the BG1 ovarian cell line that endogenously expresses full-length ER (α and β) and is stably transfected with a plasmid containing four estrogen responsive elements upstream of a luciferase reporter gene. To allow increased throughput and testing efficiency, the BG1Luc ER TA ("BG1 manual") method was adapted for quantitative high-throughput screening (BG1 qHTS) in the U.S. Tox21 testing program. The BG1 qHTS test method was used to test approximately 10,000 chemicals three times each, and concentration-response data (n=15) were analyzed to evaluate test method performance. The balanced accuracy of the BG1 qHTS test method (97% [32/33]) was determined by comparing results to ER TA performance standards for the BG1 manual method. Concordance between the BG1 manual and qHTS methods was 92% (57/62) when calculated for a larger set of non-reference chemicals tested in both methods. These data demonstrate that the performance of the BG1 qHTS is similar to the currently accepted BG1 manual method, thereby establishing the utility of the BG1 qHTS method for identifying ER active environmental chemicals.
2012年，BG1Luc4E2雌激素受体（ER）反式激活（TA）方法（BG1Luc ER TA）被美国监管机构和经济合作与发展组织认可，用于检测具有ER激动剂活性的物质。该方法现已成为美国环境保护局内分泌干扰物筛选计划一级测试组合的一部分。BG1Luc ER TA方法使用的BG1卵巢细胞系内源性表达全长ER（α和β），并稳定转染了一个在荧光素酶报告基因上游含有四个雌激素反应元件的质粒。为了提高通量和测试效率，BG1Luc ER TA（“BG1手动”）方法在美国毒物21测试计划中被改编用于定量高通量筛选（BG1 qHTS）。BG1 qHTS测试方法对约10000种化学品各进行了三次测试，并分析了浓度-反应数据（n = 15）以评估测试方法的性能。通过将结果与BG1手动方法的ER TA性能标准进行比较，确定了BG1 qHTS测试方法的平衡准确率为97%（32/33）。当对两种方法中测试的一组更大的非参考化学品进行计算时，BG1手动方法和qHTS方法之间的一致性为92%（57/62）。这些数据表明，BG1 qHTS的性能与目前认可的BG1手动方法相似，从而确立了BG1 qHTS方法在识别具有ER活性的环境化学品方面的实用性。
