Gaido K W, Leonard L S, Lovell S, Gould J C, Babaï D, Portier C J, McDonnell D P
Chemical Industry Institute of Toxicology, Research Triangle Park, North Carolina 27709, USA.
Toxicol Appl Pharmacol. 1997 Mar;143(1):205-12. doi: 10.1006/taap.1996.8069.
There is a concern that chemicals in our environment are affecting human health by disrupting normal endocrine function. Much of the concern has focused on chemicals that can interact directly with steroid hormone receptors. We have used a yeast-based assay to assess chemical interactions with the estrogen, androgen, and progesterone receptors. The yeast transformants used in this study contained the human estrogen, androgen, or progesterone receptor along with the appropriate steroid responsive elements upstream of the beta-galactosidase reporter gene. Chemicals were added to yeast cultures in doses ranging from 10(-12) to 10(-4) M and following incubation, the yeasts were then lysed and assayed for beta-galactosidase activity. Diethylstilbesterol and 17-beta estradiol were most active in the estrogen receptor assay, followed by the phytoestrogen, coumestrol. p-Nonylphenol and bisphenol A were approximately 5000- and 15,000-fold less active, respectively, than estradiol. Methoxychlor, DDT and its metabolites, o,p'-DDD, and o,p'-DDE ranged in potency from 5 to 24 X 10(6) less potent than estradiol. Testosterone and dihydrotestosterone were most potent in the androgen receptor assay, followed by estradiol and progesterone. p,p'-DDE was approximately 10(6)-fold less potent than testosterone. None of the industrial chemicals tested interacted with the progesterone receptor. These data demonstrate the utility of using yeast-based receptor assays for detecting chemical interaction with steroid receptors and these assays should serve as a useful component of an in vitro-in vivo strategy to assess the effects of chemicals on endocrine function.
人们担心我们环境中的化学物质会通过干扰正常的内分泌功能来影响人类健康。大部分担忧都集中在那些能直接与类固醇激素受体相互作用的化学物质上。我们使用了一种基于酵母的检测方法来评估化学物质与雌激素、雄激素和孕激素受体的相互作用。本研究中使用的酵母转化体包含人类雌激素、雄激素或孕激素受体，以及位于β-半乳糖苷酶报告基因上游的适当类固醇反应元件。将化学物质以10(-12)至10(-4) M的剂量添加到酵母培养物中，孵育后，裂解酵母并检测β-半乳糖苷酶活性。己烯雌酚和17-β雌二醇在雌激素受体检测中活性最高，其次是植物雌激素香豆雌酚。对壬基酚和双酚A的活性分别比雌二醇低约5000倍和15000倍。甲氧滴滴涕、滴滴涕及其代谢物邻，对'-滴滴滴和邻，对'-滴滴伊的效力比雌二醇低5至24×10(6)倍。睾酮和二氢睾酮在雄激素受体检测中效力最强，其次是雌二醇和孕激素。对，对'-滴滴伊的效力比睾酮低约10(6)倍。所测试的工业化学物质均未与孕激素受体相互作用。这些数据证明了使用基于酵母的受体检测方法来检测化学物质与类固醇受体相互作用的实用性，并且这些检测方法应作为体外-体内策略的有用组成部分，以评估化学物质对内分泌功能的影响。
