Center for Health & Bioresources, AIT Austrian Institute of Technology GmbH, Konrad Lorenz Str 24, 3430 Tulln, Austria.
Sensors (Basel). 2017 Aug 1;17(8):1760. doi: 10.3390/s17081760.
The fluorescence-based multi-analyte chip platform for the analysis of estrogenic and anti-estrogenic substances is a new in vitro tool for the high throughput screening of environmental samples. In contrast to existing tools, the chip investigates the complex action of xenoestrogens in a human cell model by characterizing protein expression. It allows for the quantification of 10 proteins secreted by MCF-7 cells, representing various biological and pathological endpoints of endocrine action and distinguishing between estrogen- and anti-estrogen-dependent secretion of proteins. Distinct protein secretion patterns of the cancer cell line after exposure to known estrogen receptor agonists ß-estradiol, bisphenol A, genistein, and nonylphenol as well as antagonists fulvestrant and tamoxifen demonstrate the potential of the chip. Stimulation of cells with Interleukin-1ß shifts concentrations of low abundant biomarkers towards the working range of the chip. In the non-stimulated cell culture, Matrix Metalloproteinase 9 (MMP-9) and Vascular Endothelial Growth Factor (VEGF) show differences upon treatment with antagonists and agonists of the estrogen receptor. In stimulated MCF-7 cells challenged with receptor agonists secretion of Monocyte Chemoattractant Protein (MCP-1), Interleukin-6 (IL-6), Rantes, and Interleukin-8 (IL-8) significantly decreases. In parallel, the proliferating effect of endocrine-disrupting substances in MCF-7 cells is assessed in a proliferation assay based on resazurin. Using ethanol as a solvent for test substances increases the background of proliferation and secretion experiments, while using dimethyl sulfoxide (DMSO) does not show any adverse effects. The role of the selected biomarkers in different physiological processes such as cell development, reproduction, cancer, and metabolic syndrome makes the chip an excellent tool for either indicating endocrine-disrupting effects in food and environmental samples, or for screening the effect of xenoestrogens on a cellular and molecular level.
基于荧光的多分析物芯片平台用于分析雌激素和抗雌激素物质，是一种用于高通量筛选环境样品的新型体外工具。与现有工具相比，该芯片通过表征蛋白质表达来研究异种雌激素在人细胞模型中的复杂作用。它允许定量 MCF-7 细胞分泌的 10 种蛋白质，代表内分泌作用的各种生物学和病理学终点，并区分雌激素和抗雌激素依赖性蛋白质分泌。在已知的雌激素受体激动剂β-雌二醇、双酚 A、染料木黄酮和壬基酚以及拮抗剂氟维司群和他莫昔芬暴露后，癌细胞系的独特蛋白质分泌模式表明了该芯片的潜力。细胞因子-1β刺激会将低丰度生物标志物的浓度推向芯片的工作范围。在未刺激的细胞培养物中，基质金属蛋白酶 9（MMP-9）和血管内皮生长因子（VEGF）在接受雌激素受体拮抗剂和激动剂治疗时表现出差异。在受受体激动剂刺激的 MCF-7 细胞中，单核细胞趋化蛋白-1（MCP-1）、白细胞介素-6（IL-6）、Rantes 和白细胞介素-8（IL-8）的分泌显著减少。同时，基于 Resazurin 的增殖测定法评估内分泌干扰物质对 MCF-7 细胞的增殖作用。使用乙醇作为测试物质的溶剂会增加增殖和分泌实验的背景，而使用二甲基亚砜（DMSO）则不会产生任何不良影响。所选生物标志物在细胞发育、生殖、癌症和代谢综合征等不同生理过程中的作用使该芯片成为一种出色的工具，可用于指示食品和环境样品中的内分泌干扰效应，或用于筛选异种雌激素对细胞和分子水平的影响。
