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Cell‐Based Bioassay to Screen Environmental Chemicals and Human Serum for Total Glucocorticogenic Activity

机译:基于细胞的生物测定,以筛选环境化学品和人血清,用于总糖钙化活性

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Glucocorticoids are steroid hormones that have systemic effects that are mediated by the glucocorticoid receptor. Environmental chemicals that disrupt glucocorticoid receptor signaling and/or glucocorticoid homeostasis could adversely affect the health of human and nonhuman vertebrates. A major challenge in identifying environmental chemicals that alter glucocorticoid receptor signaling and/or glucocorticoid homeostasis is a lack of adequate screening methods. We developed a cell-based bioassay to measure total glucocorticogenic activity (TGA) of environmental chemicals and human serum. Human MDA-MB-231 breast cancer cells were stably transfected with a luciferase reporter gene driven by 3 tandem glucocorticoid-response elements. Dose-response curves for 6 glucocorticoids and 4 non-glucocorticoid steroid hormones were generated to evaluate the specificity of the bioassay. Cells were also optimized to measure TGA of 176 structurally diverse environmental chemicals and human serum samples in a high-throughput format. Reporter activity was glucocorticoid-specific and induced 400-fold by 1 mu M dexamethasone. Furthermore, 3 of the screened chemicals (3,4,4 '-trichlorocarbanilide, isopropyl-N-phenylcarbamate, and benzothiazole derivative 2-[4-chlorophenyl]-benzothiazole) potentiated cortisol-induced glucocorticoid receptor activity. Serum TGA estimates from the bioassay were highly correlated with a cortisol enzyme-linked immunosorbent assay. The present study establishes an in vitro method to rapidly screen environmental chemicals and human serum for altered glucocorticogenic activity. Future studies can utilize this tool to quantify the joint effect of endogenous glucocorticoids and environmental chemicals. Environ Toxicol Chem 2020;00:1-10. (c) 2020 SETAC
机译:糖皮质激素是具有由糖皮质激素受体介导的系统效果的类固醇激素。破坏糖皮质激素受体信号和/或糖皮质激素稳态的环境化学品可能对人和非人类脊椎动物的健康产生不利影响。在识别改变糖皮质激素受体信号传导和/或糖皮质激素稳态的环境化学品中的主要挑战是缺乏足够的筛选方法。我们开发了一种基于细胞的生物测定,以测量环境化学品和人血清的总糖化活性(TGA)。用3串联糖皮质激素反应元素驱动的荧光素酶报告基因稳定地转染人MDA-MB-231乳腺癌细胞。产生6种糖皮质激素和4种非糖皮质激素类固醇激素的剂量反应曲线,以评估生物测定的特异性。还经过优化细胞以以高通量形式测量176个结构各种环境化学品和人血清样品的TGA。记者活性是糖皮质激素特异性的,并诱导400倍1 mu m米塞米松。此外,3种筛选的化学物质(3,4,4'-三氟甲酰胺,异丙基-N-苯基氨基甲酸酯和苯并噻唑衍生物2- [4-氯苯基] - 苯并噻唑)有序的皮质醇诱导的糖皮质激素受体活性。来自生物测定的血清TGA估计与皮质醇酶联免疫吸附测定的高度相关。本研究建立了快速筛选环境化学品和人血清的体外方法,改变糖钙化活性。未来的研究可以利用该工具量化内源性糖皮质激素和环境化学品的关节效果。环境毒素化学2020; 00:1-10。 (c)2020 Setac

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