Instituto de Investigaciones Biomédicas, Universidad de Buenos Aires-Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Medicina, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina.
Centro de Altos Estudios en Ciencias Humanas y de la Salud, Universidad Abierta Interamericana, Ciudad Autónoma de Buenos Aires, Argentina.
Environ Toxicol Chem. 2019 Dec;38(12):2719-2728. doi: 10.1002/etc.4591.
Endocrine-disrupting chemicals (EDCs) include widespread naturally occurring and synthetic substances in the environment that adversely affect humans and wildlife. Because of the increasing numbers of EDCs, screening methods and ideal biomarkers to determine EDC potencies at relevant environmental concentrations need to be drastically improved. Soluble guanylyl cyclase α1 subunit (sGCα1) is an abundant cytosolic protein ubiquitously expressed in most tissues. We previously showed that sGCα1 is specifically and highly up-regulated by estrogen (E2) in vivo and in vitro, even though it lacks estrogen-responsive elements. The aim of the present study was to evaluate sGCα1 protein expression as a potential marker for xenoestrogenic EDC exposure in the E2-responsive lactosomatotroph-derived pituitary cell line GH3. Cells were incubated with a wide variety of EDCs such as heavy metals and a metalloid, synthetic E2 derivatives, plastic byproducts, and pesticides at a range of doses including those with proven xenoestrogenic activity. We demonstrated that E2 increased sGCα1 expression in GH3 cells as well as in other E2-responsive tumor cell lines. Moreover, this effect was fully dependent on estrogen receptor (ER) activation. Importantly, sGCα1 protein levels were strongly up-regulated by all the EDCs tested, even by those exhibiting low or null ER binding capacity. We provide evidence that the in vitro sGCα1 protein assay may be a very sensitive and powerful tool to identify compounds with estrogenic activity, which could improve current mammalian-based screening methods. Environ Toxicol Chem 2019;38:2719-2728. © 2019 SETAC.
内分泌干扰化学物质（EDCs）包括环境中广泛存在的天然和合成物质，它们会对人类和野生动物造成不良影响。由于 EDCs 的数量不断增加，需要大幅改进筛选方法和理想的生物标志物，以确定相关环境浓度下的 EDC 效力。可溶性鸟苷酸环化酶 α1 亚基（sGCα1）是一种丰富的细胞质蛋白，在大多数组织中广泛表达。我们之前表明，sGCα1 被体内和体外的雌激素（E2）特异性和高度上调，尽管它缺乏雌激素反应元件。本研究的目的是评估 sGCα1 蛋白表达作为一种潜在的标记物，用于评估雌激素反应性乳垂体衍生 GH3 细胞系中 EDC 暴露的外源性雌激素。将细胞与各种 EDC 孵育，如重金属和类金属、合成 E2 衍生物、塑料副产品和杀虫剂，剂量范围包括已证明具有外源性雌激素活性的剂量。我们证明 E2 增加了 GH3 细胞以及其他雌激素反应性肿瘤细胞系中的 sGCα1 表达。此外，这种作用完全依赖于雌激素受体（ER）的激活。重要的是，所有测试的 EDC 都强烈地上调了 sGCα1 蛋白水平，即使是那些表现出低或无 ER 结合能力的 EDC 也是如此。我们提供的证据表明，体外 sGCα1 蛋白测定可能是一种非常敏感和强大的工具，可用于识别具有雌激素活性的化合物，从而改进当前基于哺乳动物的筛选方法。环境毒理化学 2019；38:2719-2728. © 2019 SETAC。
