Marine Science Institute, University of Texas at Austin, 750 Channel View Drive, Port Aransas, TX 78373, United States.
Marine Science Institute, University of Texas at Austin, 750 Channel View Drive, Port Aransas, TX 78373, United States.
Steroids. 2019 Sep;149:108415. doi: 10.1016/j.steroids.2019.05.007. Epub 2019 May 30.
A variety of pesticides including vinclozolin, its metabolite M2 (3',5'-dichloro-2-hydroxy-2-methylbut-3-enanilide), and prochloraz have been shown to exert antiandrogenic effects in animal models by competing with androgen binding to nuclear androgen receptors (nAR) and decreasing transcription of androgen-responsive genes. However, it is not known whether these pesticide antiandrogens also interfere with rapid (often described as nongenomic, nonclassical) androgen actions mediated by membrane androgen receptors (mARs). We recently discovered that ZIP9, a member of the zinc transporter ZIP (SLC39A) family, is a specific, high-affinity mAR that mediates rapid testosterone-dependent signaling, zinc influx, and apoptosis in breast and prostate cancer cell lines. Possible disruption by prochloraz, vinclozolin, and M2 of androgen actions through this mAR was investigated in vitro in PC-3 prostate cancer cells (nAR-) over expressing human ZIP9 (PC3-ZIP9 cells). Single-point competitive binding assays showed 1 μM and 10 μM concentrations of all three pesticides displaced specific [H]-testosterone binding to PC3-ZIP9 cell membranes with binding affinities <10% that of testosterone. The pesticides also exerted antiandrogen actions through ZIP9. Co-treatments with 100 nM prochloraz, vinclozolin and M2 blocked or attenuated the 20 nM testosterone-induced increases in apoptosis, intracellular free zinc levels, and expression of the proapoptotic gene, Bax. Prochloraz also attenuated testosterone activation of MAPkinase. The finding that prochloraz, vinclozolin and M2 are effective competitors of [H]-testosterone binding to ZIP9 and block testosterone actions mediated through ZIP9 in vitro at nanomolar concentrations suggests that androgen functions mediated by ZIP9 are also susceptible to disruption by pesticide antiandrogens with potential adverse effects on human health.
包括烯效唑、其代谢物 M2（3'，5'-二氯-2-羟基-2-甲基丁-3-烯酰苯胺）和百菌清在内的多种农药已被证明在动物模型中具有抗雄激素作用，其通过与核雄激素受体（nAR）竞争结合并降低雄激素反应基因的转录来发挥作用。然而，目前尚不清楚这些农药抗雄激素是否也会干扰由膜雄激素受体（mAR）介导的快速（通常描述为非基因组、非经典）雄激素作用。我们最近发现，锌转运体 ZIP（SLC39A）家族的成员 ZIP9 是一种特异性、高亲和力的 mAR，可介导乳腺癌和前列腺癌细胞系中快速的睾酮依赖性信号转导、锌内流和细胞凋亡。在过表达人 ZIP9（PC3-ZIP9 细胞）的前列腺癌细胞系（nAR-）中，体外研究了百菌清、烯效唑和 M2 可能通过这种 mAR 对雄激素作用的干扰。单点竞争结合测定显示，三种农药的 1µM 和 10µM 浓度均能置换 PC3-ZIP9 细胞膜上特异性[H]-睾酮结合，其结合亲和力低于睾酮的 10%。这些农药还通过 ZIP9 发挥抗雄激素作用。用 100nM 百菌清、烯效唑和 M2 共同处理可阻断或减弱 20nM 睾酮诱导的细胞凋亡、细胞内游离锌水平和促凋亡基因 Bax 的表达增加。百菌清还减弱了睾酮激活 MAPkinase。百菌清、烯效唑和 M2 有效竞争[H]-睾酮与 ZIP9 结合，并在纳摩尔浓度下阻断 ZIP9 介导的睾酮作用的发现表明，ZIP9 介导的雄激素功能也易受农药抗雄激素的干扰，可能对人类健康产生不利影响。
