Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor-reporter gene assays
Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor-reporter gene assays
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DOI：
10.1139/cjpp-74-2-216
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发表时间：
1996-02-01
影响因子：
2.1
通讯作者：
Zacharewski, T
中科院分区：
文献类型：
--
作者：
Balaguer, P;Joyeux, A;Zacharewski, T
体外重组受体报告基因分析已经被用来评估和排序怀疑具有雌激素和(或)芳烃受体(AhR)介导的活性的化学品和复杂混合物的效力。环境雌激素(E(2))生物测定由Gal4-人雌激素受体嵌合结构(Gal4-Hego)和Gal4调节的荧光素酶报告基因(17m5-G-Luc)组成，它们已稳定整合到HeLa细胞中。经1 NM 17β-雌二醇处理后，荧光素酶报告基因活性增加了10倍，检测下限约为5 pg/ml。AhR生物检测使用瞬时转染二恶英反应元件调节的荧光素酶报告基因的HEPA 1c1c7野生型细胞。这些测定被用来评估柚皮苷、阿特拉津、西马津以及纸浆和造纸厂等复杂混合物的雌激素和二恶英样活性。黑液和城市空气颗粒物。用纯抗雌激素ICI164、384和体外凝胶滞留试验证实了这些化合物和复杂混合物的活性。这项研究的结果证明了体外重组受体报告基因分析在鉴定和评估化学品和复杂混合物的雌激素和二恶英样活性方面的有效性。
In vitro recombinant receptor-reporter gene assays have been used to assess and rank the potency of chemicals and complex mixtures suspected of possessing estrogen and (or) aryl hydrocarbon receptor (AhR) mediated activity. The environmental estrogen (E(2)) bioassay consists of a Gal4-human estrogen receptor chimeric construct (Gal4-HEGO) and a Gal4-regulated luciferase reporter gene (17m5-G-Luc) that have been stably integrated into HeLa cells. The assay exhibits 10-fold induction in luciferase reporter gene activity following treatment with 1 nM 17 beta-estradiol and has a detection limit of approximately 5 pg of 17 beta-estradiol/mL. The AhR bioassay uses Hepa 1c1c7 wild-type cells transiently transfected with a dioxin response element regulated luciferase reporter gene. These assays were used to assess the estrogen and dioxin-like activities of naringenin, atrazine, and simazine and complex mixtures such as pulp and paper mill. black liquor and urban air particulates. The activities of these chemicals and complex mixtures are confirmed using the pure antiestrogen ICI 164,384 and in in vitro gel retardation assays. Results of this study demonstrate the utility of in vitro recombinant receptor-reporter gene assays in identifying and assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures.