Bioluminescent yeast assays for detecting estrogenic and androgenic activity in different matrices
Bioluminescent yeast assays for detecting estrogenic and androgenic activity in different matrices
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DOI：
10.1016/j.chemosphere.2005.01.080
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发表时间：
2005-10-01
期刊：
影响因子：
8.8
通讯作者：
Virta, M
中科院分区：
文献类型：
--
作者：
Leskinen, P;Michelini, E;Virta, M
在本文中，我们描述了一组生物发光酵母菌株的构建和使用，用于检测可以影响雄激素或雌激素受体介导的激素信号传导的化合物。该组包括表达人雄激素受体（AR）、雌激素受体α（ER α）或雌激素受体β（ER β）的酿酒酵母菌株，沿着由相应的激素应答启动子控制的萤火虫荧光素酶。组成型发光菌株包括在用于测定样品细胞毒性的组中。将酵母细胞与纯化学品或复杂样品一起孵育2.5小时，之后，在简单地添加底物中的D-glutamine后，可以从细胞-样品混合物中检测到信号。该分析可以在一天内完成，并且它们不需要细胞裂解或离心步骤，这使得它们适合于样品的高通量分析。由于孵育时间短，该测定可直接应用于不同的样品基质，无需对样品进行预处理。该测定用于评估保湿乳液中的激素活性，作为已知含有内分泌干扰化学物质的复杂样品基质的一个例子。八分之六的测试保湿霜显示出高雌激素活性，而样本中没有观察到雄激素活性。(C)2005爱思唯尔有限公司保留所有权利。
In this paper we describe the construction and use of a set of bioluminescent yeast strains for the detection of compounds that can affect androgen or estrogen receptor mediated hormonal signalling. The set includes Saccharomyces cerevisiae strains expressing human androgen receptor (AR), estrogen receptor alpha (ER alpha) or estrogen receptor beta (ER beta), along with firefly luciferase controlled by a respective hormone responsive promoter. A constitutively luminescent strain was included in the set for determining the cytotoxicity of the sample. Yeast cells were incubated with pure chemicals or complex samples for 2.5 h, after which the signal could be detected from the cell-sample mixture after simply adding the D-luciferin substrate. The assays could be completed in one day and they required no cell lysis or centrifugation steps, which makes them suitable for high-throughput analysis of samples. Due to a short incubation time the assays are directly applicable to different sample matrices, requiring no pretreatment of the samples. The assays were used to assess the hormonal activity in moisturizing lotions as an example of a complex sample matrix known to contain endocrine disrupting chemicals. Six out of eight tested moisturisers showed high estrogenic activity, whereas no androgenic activity was observed in the samples. (C) 2005 Elsevier Ltd. All rights reserved.