Suppr超能文献

AroER 三屏检测法是一种生物学相关的检测方法,用于检测内分泌干扰物对芳香化酶和/或雌激素受体活性的调节作用。

AroER tri-screen is a biologically relevant assay for endocrine disrupting chemicals modulating the activity of aromatase and/or the estrogen receptor.

机构信息

Department of Cancer Biology, Beckman Research Institute of the City of Hope, Duarte, California 91010.

出版信息

Toxicol Sci. 2014 May;139(1):198-209. doi: 10.1093/toxsci/kfu023. Epub 2014 Feb 4.

Abstract

Endocrine disrupting chemicals (EDCs) interfere with the biosynthesis, metabolism, and functions of steroid hormones, including estrogens and androgens. Aromatase enzyme converts androgen to estrogen. Thus, EDCs against aromatase significantly impact estrogen- and/or androgen-dependent functions, including the development of breast cancer. The current study aimed to develop a biologically relevant cell-based high-throughput screening assay to identify EDCs that act as aromatase inhibitors (AIs), estrogen receptor (ER) agonists, and/or ER antagonists. The AroER tri-screen assay was developed by stable transfection of ER-positive, aromatase-expressing MCF-7 breast cancer cells with an estrogen responsive element (ERE) driven luciferase reporter plasmid. The AroER tri-screen can identify: estrogenic EDCs, which increase luciferase signal without 17β-estradiol (E2); anti-estrogenic EDCs, which inhibit the E2-induced luciferase signal; and AI-like EDCs, which suppress a testosterone-induced luciferase signal. The assay was first optimized in a 96-well plate format and then miniaturized into a 1536-well plate format. The AroER tri-screen was demonstrated to be suitable for high-throughput screening in the 1536-well plate format, with a 6.9-fold signal-to-background ratio, a 5.4% coefficient of variation, and a screening window coefficient (Z-factor) of 0.78. The assay suggested that bisphenol A (BPA) functions mainly as an ER agonist. Results from screening the 446 drugs in the National Institutes of Health Clinical Collection revealed 106 compounds that modulated ER and/or aromatase activities. Among these, two AIs (bifonazole and oxiconazole) and one ER agonist (paroxetine) were confirmed through alternative aromatase and ER activity assays. These findings indicate that AroER tri-screen is a useful high-throughput screening system for identifying ER ligands and aromatase-inhibiting chemicals.

摘要

内分泌干扰化学物质 (EDCs) 干扰甾体激素(包括雌激素和雄激素)的生物合成、代谢和功能。芳香酶将雄激素转化为雌激素。因此,针对芳香酶的 EDC 会显著影响雌激素和/或雄激素依赖性功能,包括乳腺癌的发展。本研究旨在开发一种具有生物学相关性的基于细胞的高通量筛选测定法,以鉴定作为芳香酶抑制剂 (AIs)、雌激素受体 (ER) 激动剂和/或 ER 拮抗剂的 EDC。AroER 三联筛选测定法通过将具有雌激素反应元件 (ERE) 驱动的荧光素酶报告质粒的 ER 阳性、芳香酶表达 MCF-7 乳腺癌细胞进行稳定转染而开发。AroER 三联筛选可以鉴定:增加荧光素酶信号而无需 17β-雌二醇 (E2) 的雌激素性 EDC;抑制 E2 诱导的荧光素酶信号的抗雌激素性 EDC;以及类似 AI 的 EDC,其抑制睾酮诱导的荧光素酶信号。该测定法首先在 96 孔板格式中进行优化,然后小型化为 1536 孔板格式。证明 AroER 三联筛选适用于 1536 孔板格式的高通量筛选,具有 6.9 倍的信号与背景比、5.4%的变异系数和 0.78 的筛选窗口系数 (Z 因子)。该测定法表明双酚 A (BPA) 主要作为 ER 激动剂发挥作用。对国立卫生研究院临床收藏中的 446 种药物进行筛选的结果表明,有 106 种化合物调节了 ER 和/或芳香酶活性。其中,两种 AI(比佛拉唑和奥昔康唑)和一种 ER 激动剂(帕罗西汀)通过替代芳香酶和 ER 活性测定得到证实。这些发现表明,AroER 三联筛选是一种用于鉴定 ER 配体和芳香酶抑制性化学物质的有用高通量筛选系统。

相似文献

2
AroER tri-screen™ is a novel functional assay to estimate both estrogenic and estrogen precursor activity of chemicals or biological specimens.
Breast Cancer Res Treat. 2015 Jun;151(2):335-45. doi: 10.1007/s10549-015-3398-z. Epub 2015 May 12.
3
Cell-Based High-Throughput Screening for Aromatase Inhibitors in the Tox21 10K Library.
Toxicol Sci. 2015 Oct;147(2):446-57. doi: 10.1093/toxsci/kfv141. Epub 2015 Jul 3.
5
MCF-7aro/ERE, a novel cell line for rapid screening of aromatase inhibitors, ERalpha ligands and ERRalpha ligands.
Biochem Pharmacol. 2008 Jul 15;76(2):208-15. doi: 10.1016/j.bcp.2008.04.011. Epub 2008 May 1.
6
Identification of putative estrogen receptor-mediated endocrine disrupting chemicals using QSAR- and structure-based virtual screening approaches.
Toxicol Appl Pharmacol. 2013 Oct 1;272(1):67-76. doi: 10.1016/j.taap.2013.04.032. Epub 2013 May 23.
7
Identification of nonmonotonic concentration-responses in Tox21 high-throughput screening estrogen receptor assays.
Toxicol Appl Pharmacol. 2022 Oct 1;452:116206. doi: 10.1016/j.taap.2022.116206. Epub 2022 Aug 19.
8
Currently used pesticides and their mixtures affect the function of sex hormone receptors and aromatase enzyme activity.
Toxicol Appl Pharmacol. 2013 Oct 15;272(2):453-64. doi: 10.1016/j.taap.2013.06.028. Epub 2013 Jul 16.
9
Endocrine-Disrupting Chemicals (EDCs): In Vitro Mechanism of Estrogenic Activation and Differential Effects on ER Target Genes.
Environ Health Perspect. 2013 Apr;121(4):459-66. doi: 10.1289/ehp.1205951. Epub 2013 Feb 5.

引用本文的文献

1
Use of Tox21 screening data to profile PFAS bioactivities on nuclear receptors, cellular stress pathways, and cytochrome p450 enzymes.
J Hazard Mater. 2024 Jul 15;473:134642. doi: 10.1016/j.jhazmat.2024.134642. Epub 2024 May 17.
4
A Fluorometric CYP19A1 (Aromatase) Activity Assay in Live Cells.
ChemMedChem. 2021 Sep 16;16(18):2845-2850. doi: 10.1002/cmdc.202100326. Epub 2021 Jul 5.
5
Paroxetine-Overview of the Molecular Mechanisms of Action.
Int J Mol Sci. 2021 Feb 7;22(4):1662. doi: 10.3390/ijms22041662.
6
A Gene Expression Biomarker Identifies Chemical Modulators of Estrogen Receptor α in an MCF-7 Microarray Compendium.
Chem Res Toxicol. 2021 Feb 15;34(2):313-329. doi: 10.1021/acs.chemrestox.0c00243. Epub 2021 Jan 6.
7
Comparing Machine Learning Models for Aromatase (P450 19A1).
Environ Sci Technol. 2020 Dec 1;54(23):15546-15555. doi: 10.1021/acs.est.0c05771. Epub 2020 Nov 19.
8
Comparison of Machine Learning Models for the Androgen Receptor.
Environ Sci Technol. 2020 Nov 3;54(21):13690-13700. doi: 10.1021/acs.est.0c03984. Epub 2020 Oct 21.
9
Computational Study of Drugs Targeting Nuclear Receptors.
Molecules. 2020 Apr 1;25(7):1616. doi: 10.3390/molecules25071616.
10
Towards a Treatment for Gulf War Illness: A Consensus Docking Approach.
Mil Med. 2020 Jan 7;185(Suppl 1):554-561. doi: 10.1093/milmed/usz299.

本文引用的文献

1
Androgen (dihydrotestosterone)-mediated regulation of food intake and obesity in female mice.
J Steroid Biochem Mol Biol. 2013 Nov;138:100-6. doi: 10.1016/j.jsbmb.2013.04.001. Epub 2013 May 7.
2
Improving the human hazard characterization of chemicals: a Tox21 update.
Environ Health Perspect. 2013 Jul;121(7):756-65. doi: 10.1289/ehp.1205784. Epub 2013 Apr 19.
3
Bisphenol S disrupts estradiol-induced nongenomic signaling in a rat pituitary cell line: effects on cell functions.
Environ Health Perspect. 2013 Mar;121(3):352-8. doi: 10.1289/ehp.1205826. Epub 2013 Jan 17.
4
Obesity and cancer risk: evidence, mechanisms, and recommendations.
Ann N Y Acad Sci. 2012 Oct;1271(1):37-43. doi: 10.1111/j.1749-6632.2012.06750.x.
5
Analysis of estrogen receptor isoforms and variants in breast cancer cell lines.
Exp Ther Med. 2011 May;2(3):537-544. doi: 10.3892/etm.2011.226. Epub 2011 Mar 10.
6
NCBI Reference Sequences (RefSeq): current status, new features and genome annotation policy.
Nucleic Acids Res. 2012 Jan;40(Database issue):D130-5. doi: 10.1093/nar/gkr1079. Epub 2011 Nov 24.
7
Chemical genomics profiling of environmental chemical modulation of human nuclear receptors.
Environ Health Perspect. 2011 Aug;119(8):1142-8. doi: 10.1289/ehp.1002952. Epub 2011 May 4.
8
Endocrine disruptors: from endocrine to metabolic disruption.
Annu Rev Physiol. 2011;73:135-62. doi: 10.1146/annurev-physiol-012110-142200.
9
The HDAC inhibitor LBH589 (panobinostat) is an inhibitory modulator of aromatase gene expression.
Proc Natl Acad Sci U S A. 2010 Jun 15;107(24):11032-7. doi: 10.1073/pnas.1000917107. Epub 2010 Jun 1.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验