Evaluation of a recombinant yeast cell estrogen screening assay.
Evaluation of a recombinant yeast cell estrogen screening assay.
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评估重组酵母细胞雌激素筛查测定法。
DOI：
10.1289/ehp.97105734
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发表时间：
1997-07
影响因子：
10.4
通讯作者：
Sauer MJ
中科院分区：
文献类型：
--
作者：
Coldham NG;Dave M;Sivapathasundaram S;McDonnell DP;Connor C;Sauer MJ
据报告，源自植物和环境的各种结构的化学品具有激素活性。人类可能会明显接触到这些物质，因此有必要开发敏感的筛选方法，以量化和评估对公众的风险。用编码人雌激素受体和与报告基因连接的雌激素应答启动子的质粒转化的酵母细胞被评价用于筛选具有雌激素活性的化合物。通过参考使用重组酵母细胞、MCF-7人乳腺癌细胞和青春期前小鼠子宫营养生物测定得出的17 β-雌二醇（E2）校准曲线，评价对雌激素的相对敏感性。重组酵母细胞生物测定法（RCBA）对E2的敏感性分别比MCF-7细胞和子宫营养测定法高出两个和五个数量级。使用RCBA测定了53种化学物质的雌激素效能，包括类固醇激素、合成雌激素、环境污染物和植物雌激素。相对于E2（100），RCBA产生的效价值包括雌酮（9.6）、己烯雌酚（74.3）、他莫昔芬（0.0047）、α-玉米赤霉醇（1.3）、雌马酚（0.085）、4-壬基苯酚（0.005）和邻苯二甲酸丁基苄酯（0.0004），这些值与文献值相似，但通常高于子宫营养测定产生的值。灵敏度高，没有测试化合物的生物转化，易于使用，并测量抗雌激素活性的可能性是重要的属性，认为RCBA在筛选潜在的异种雌激素，以评估对人类，野生动物和环境的风险的适用性。
A wide range of chemicals with diverse structures derived from plant and environmental origins are reported to have hormonal activity. The potential for appreciable exposure of humans to such substances prompts the need to develop sensitive screening methods to quantitate and evaluate the risk to the public. Yeast cells transformed with plasmids encoding the human estrogen receptor and an estrogen responsive promoter linked to a reporter gene were evaluated for screening compounds for estrogenic activity. Relative sensitivity to estrogens was evaluated by reference to 17 beta-estradiol (E2) calibration curves derived using the recombinant yeast cells, MCF-7 human breast cancer cells, and a prepubertal mouse uterotrophic bioassay. The recombinant yeast cell bioassay (RCBA) was approximately two and five orders of magnitude more sensitive to E2 than MCF-7 cells and the uterotrophic assay, respectively. The estrogenic potency of 53 chemicals, including steroid hormones, synthetic estrogens, environmental pollutants, and phytoestrogens, was measured using the RCBA. Potency values produced with the RCBA relative to E2 (100) included estrone (9.6), diethylstilbestrol (74.3), tamoxifen (0.0047), alpha-zearalanol (1.3), equol (0.085), 4-nonylphenol (0.005), and butylbenzyl phathalate (0.0004), which were similar to literature values but generally higher than those produced by the uterotrophic assay. Exquisite sensitivity, absence of test compound biotransformation, ease of use, and the possibility of measuring antiestrogenic activity are important attributes that argue for the suitability of the RCBA in screening for potential xenoestrogens to evaluate risk to humans, wildlife, and the environment.