Rapid yeast estrogen bioassays stably expressing human estrogen receptors α and β, and green fluorescent protein:: a comparison of different compounds with both receptor types
Rapid yeast estrogen bioassays stably expressing human estrogen receptors α and β, and green fluorescent protein:: a comparison of different compounds with both receptor types
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DOI：
10.1016/j.jsbmb.2004.03.118
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发表时间：
2004-07-01
影响因子：
4.1
通讯作者：
Hoogenboom, RLAP
中科院分区：
文献类型：
--
作者：
Bovee, TFH;Helsdingen, RJR;Hoogenboom, RLAP
以前，我们描述了一个快速酵母生物测定的建设稳定表达人雌激素受体α（hER α）和酵母增强绿色荧光蛋白（yEGFP）在响应雌激素。在本研究中，通过测试一系列雌激素化合物进一步研究了该测定法的性质。此外，基于人雌激素受体β（hER β）的稳定表达开发了类似的测定。当暴露于17 β-雌二醇时，ER β细胞传感器的最大转录活性仅为ER α观察到的活性的约40%，但达到半最大激活的浓度（EC 50）低约5倍。相对雌激素效能（REP），定义为17 β-雌二醇的EC 50与化合物的EC 50之间的比率，合成激素二烯雌酚，己雌酚，尤其是美雌醇与ER α的比值更高，而DES与ER β的比值略高。孕激素孕酮和醋酸甲羟孕酮无反应，而雄激素睾酮反应很弱。合成代谢剂19-去甲睾酮与雌激素受体α（而非β）表现出明显的剂量相关反应。植物雌激素香豆雌酚、染料木黄酮、染料木苷、大豆苷元、大豆苷和柚皮素对ERP的作用相对较强。雌激素受体α的雌激素活性排序为：17 β-雌二醇> 8-异戊烯基柚皮素>香豆雌酚>玉米赤霉烯酮>染料木黄酮>柚皮素。雌激素对雌激素受体的影响大小依次为：17 β-雌二醇>香豆雌酚>染料木黄酮>玉米赤霉烯酮> 8-异戊烯基柚皮素>大豆苷元>柚皮素>染料木黄酮>大豆苷元。啤酒花雌激素8-prenylnaringenin与ER α相对更有效。这些数据表明，新开发的生物测定是有价值的工具，快速和高通量筛选雌激素活性。(C)2004 Elsevier Ltd.保留所有权利。
Previously, we described the construction of a rapid yeast bioassay stably expressing human estrogen receptor alpha (hERalpha) and yeast enhanced green fluorescent protein (yEGFP) in response to estrogens. In the present study, the properties of this assay were further studied by testing a series of estrogenic compounds. Furthermore, a similar assay was developed based on the stable expression of human estrogen receptor beta (hERbeta). When exposed to 17beta-estradiol, the maximum transcriptional activity of the ERbeta cytosensor was only about 40% of the activity observed with ERalpha, but the concentration where half-maximal activation is reached (EC50), was about five times lower. The relative estrogenic potencies (REP), defined as the ratio between the EC50 of 17beta-estradiol and the EC50 of the compound, of the synthetic hormones dienestrol, hexestrol and especially mestranol were higher with ERalpha, while DES was slightly more potent with ERbeta. The gestagens progesterone and medroxyprogesterone- acetate showed no response, whereas the androgen testosterone showed a very weak response. The anabolic agent, 19-nortestosterone showed a clear dose-related response with estrogen receptor alpha but not beta. The phytoestrogens coumestrol, genistein, genistin, daidzein, daidzin and naringenin were relatively more potent with ERP. Ranking of the estrogenic potency with ERalpha was: 17beta-estradiol much greater than 8-prenylnaringenin > coumestrol > zearalenone much greater than genistein much greater than genistin > naringenin. The ranking with the ERp was: 17beta-estradiol much greater than cournestrol > genistein > zearalenone > 8-prenylnaringen much greater than daidzein > naringenin > genistin much greater than daidzin. The hop estrogen 8-prenylnaringenin is relatively more potent with ERalpha. These data show that the newly developed bioassays are valuable tools for the rapid and high-throughput screening for estrogenic activity. (C) 2004 Elsevier Ltd. All rights reserved.