Assessment of estrogenic chemicals using an estrogen receptor α (ERα)- and ERβ-mediated reporter gene assay in fish
Assessment of estrogenic chemicals using an estrogen receptor α (ERα)- and ERβ-mediated reporter gene assay in fish
复制标题
DOI：
10.1007/s00227-005-0206-z
复制
发表时间：
2006-04-01
期刊：
影响因子：
2.4
通讯作者：
Abe, SI
中科院分区：
文献类型：
--
作者：
Kitano, T;Koyanagi, T;Abe, SI
使用脊椎动物细胞系或酵母的体外报告基因测定已用于评估雌激素化学品。然而，鱼类雌激素受体α（ER α）和ER β介导的报告基因系统尚未开发。在本研究中，我们开发了一种ER α和ER β介导的报告基因在鱼和估计雌激素活性的17 β-雌二醇（E2），p-壬基酚（NP），双酚A（BPA），p，p '-DDE，和染料木素（Gen）使用体外报告分析。在E2或Gen给药条件下，ER α或ER β表达质粒均能强烈诱导该活性，而在NP给药条件下，ER α表达质粒能显著诱导该活性，而ER β表达质粒则无此活性。在BPA或p，p '-DDE的作用下，ER α表达质粒比ER β表达质粒更能诱导细胞的活性。这些结果表明，在体外检测的雌激素化合物中，通过ER α和ER β的活性存在明显差异。因此，体外报告基因分析为阐明雌激素类化合物在鱼类中的作用机制提供了一个很好的系统。
In vitro reporter gene assays using vertebrate cell lines or yeast have been used for assessment of the estrogenic chemicals. However, estrogen receptor alpha (ER alpha)- and ER beta-mediated reporter gene system in fish has yet to be developed. In the present study, we developed an ER alpha- and ER beta-mediated reporter gene assay in fish and estimated estrogenic activities of 17 beta-estradiol (E2), p-nonylphenol (NP), bisphenol A (BPA), p,p'-DDE, and genistein (Gen) using the in vitro reporter assay. The activity was intensely induced by transfection with either ER alpha or ER beta expression plasmid under E2 or Gen administration, whereas it was significantly induced by transfection with ER alpha expression plasmid, but not with ER beta expression plasmid under NP administration. On the other hand, the activity was induced more intensely by transfection with ER alpha expression plasmid than ER beta expression plasmid under BPA or p,p'-DDE administration. These results indicate that there are obvious differences in the activity through ER alpha and ER beta among the estrogenic chemicals examined in vitro. Thus, the in vitro reporter assay provides an excellent system for elucidating the action mechanism of estrogenic chemicals in fishes.