School of Public Health, Xuzhou Medical College, Xuzhou, 84 West Huai-hai Road, Xuzhou, Jiangsu 221002, China.
Toxicology. 2013 Sep 15;311(3):178-83. doi: 10.1016/j.tox.2013.06.011. Epub 2013 Jul 4.
To clarify whether the mechanism of androgen receptor (AR) antagonism of the pyrethroid pesticide cypermethrin associates with the interactions between the AR and corepressors silencing mediator for thyroid hormone receptors (SMRT) and nuclear receptor corepressor (NCoR), we have developed the mammalian two-hybrid assays. The AR N-terminal domain 1-660 amino acid residues were subcloned into the plasmid pVP16 to construct VP16-ARNTD. The C-terminal receptor interaction domains (RIDs) of SMRT and NCoR were used to construct pM-SMRT and pM-NCoR. The constructed vectors pVP16-ARNTD, pM-SMRT or pM-NCoR, the reporter pG5CAT and the control pCMVβ were cotranfected into the CV-1 cells. The cells were treated with cypermethrin at the indicated concentrations. The AR N terminus interacted with RIDs of SMRT and NCoR. The interactions between the AR and corepressors SMRT and NCoR were enhanced by cypermethrin, and the significant enhancement was detected at the concentration of 10(-5)M. The mammalian two-hybrid assays demonstrate the utility to detect the interactions of the AR with SMRT and NCoR. Cypermethrin functions as an anti-androgen by enhancing the associations of the AR with SMRT and NCoR. We provide a novel mechanism in anti-androgen action of cypermethrin associated with the recruitment of SMRT and NCoR to AR.
为了阐明除虫菊酯类农药氯菊酯的雄激素受体（AR）拮抗作用的机制是否与 AR 与甲状腺激素受体沉默介质（SMRT）和核受体辅抑制因子（NCoR）的核心抑制因子之间的相互作用有关，我们已经开发了哺乳动物双杂交测定法。将 AR N 端结构域 1-660 个氨基酸残基亚克隆到质粒 pVP16 中，构建 VP16-ARNTD。SMRT 和 NCoR 的 C 端受体相互作用结构域（RIDs）用于构建 pM-SMRT 和 pM-NCoR。构建的载体 pVP16-ARNTD、pM-SMRT 或 pM-NCoR、报告基因 pG5CAT 和对照 pCMVβ 被共转染到 CV-1 细胞中。用指示浓度的氯菊酯处理细胞。AR N 端与 SMRT 和 NCoR 的 RID 相互作用。氯菊酯增强了 AR 与核心抑制因子 SMRT 和 NCoR 之间的相互作用，在 10（-5）M 的浓度下检测到明显的增强。哺乳动物双杂交测定法证明了检测 AR 与 SMRT 和 NCoR 相互作用的有效性。氯菊酯通过增强 AR 与 SMRT 和 NCoR 的关联而发挥抗雄激素作用。我们提供了一种与 SMRT 和 NCoR 募集相关的氯菊酯抗雄激素作用的新机制。
