Reporter cell lines to study the estrogenic effects of xenoestrogens
Reporter cell lines to study the estrogenic effects of xenoestrogens
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DOI：
10.1016/s0048-9697(99)00178-3
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发表时间：
1999-08-15
影响因子：
9.8
通讯作者：
Casellas, C
中科院分区：
文献类型：
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作者：
Balaguer, P;François, F;Casellas, C
为了表征化学品的雌激素活性，我们在稳定转染的MCF-7和HeLa细胞中建立了互补的体外重组受体-报告基因测定。表达内源性雌激素受体α（ER α）的MCF-7细胞仅用雌激素调节的荧光素酶基因稳定转染。这些细胞能够检测结合ER α或干扰ER α介导的基因表达诱导的化合物。此外，用ER α或ER β构建体以及雌激素调节的荧光素酶基因或嵌合GAL 4-ER α受体和相应的荧光素酶报告基因转染不表达内源性ER的HeLa细胞。最后，我们测试了这四个细胞模型作为工具来检查几个潜在的外源性雌激素的雌激素活性，并检测废水污水处理流出物中的雌激素活性。在所有的模型中，壬基酚混合物（NPm），4 n-壬基酚（4 nNP），2，4 '-DDE，4，4'-DDE和废水污水处理流出物是活跃的，而PCB混合物（Aroclor 1254），PCB 77，阿特拉津和林丹（γ-六氯环己烷）是不活跃的。二恶英部分激活MCF-7细胞中的雌激素受体，而在HeLa衍生的细胞系中，它降低了雌激素诱导的荧光素酶表达。(C)1999 Elsevier Science B. V.保留所有权利。
In order to characterize the estrogenic activity of chemicals, we established complementary in vitro recombinant receptor-reporter gene assays in stably transfected MCF-7 and HeLa cells. MCF-7 cells which express the endogenous estrogen receptor alpha (ER alpha) were stably transfected with only an estrogen-regulated luciferase gene. These cells enable the detection of compounds which bind to ER alpha or interfere with the induction of ER alpha mediated gene expression. Furthermore, HeLa cells, which do not express endogenous ERs, were transfected with an ER alpha or an ER beta construct together with an estrogen-regulated luciferase gene, or a chimeric GAL4-ER alpha receptor and the corresponding luciferase reporter gene. Finally, we tested these four cellular models as tools to check the estrogenic activities of several potential xenoestrogens and to detect estrogenic activity in wastewater sewage treatment effluents. In all of the models, nonylphenol mixture (NPm), 4n-nonylphenol (4nNP), 2,4'-DDE, 4,4'-DDE and wastewater sewage treatment effluent were active, while PCB mixture (Aroclor 1254), PCB 77, atrazine and lindane (gamma hexachlorocyclohexane) were inactive. Dioxin partially activates the estrogen receptor in MCF-7 cells while in HeLa-derived cell lines, it decreased the estrogenic-induced expression of luciferase. (C) 1999 Elsevier Science B.V. All rights reserved.