Physiologically Relevant Estrogen Receptor Alpha Pathway Reporters for Single-Cell Imaging-Based Carcinogenic Hazard Assessment of Estrogenic Compounds
| Autor: | Tineke van den Hoorn, Jan Willem van der Laan, Peter Bouwman, Annabel Goudriaan, Sylvia E. Le Dévédec, Wanda van der Stel, Bob van de Water, Britt Duijndam |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
AcademicSubjects/SCI01040 Cell Estrogen receptor Toxicology live-cell imaging 03 medical and health sciences 0302 clinical medicine Cell Line Tumor medicine Estrogen Receptor beta Humans fluorescent reporters Endocrine Toxicology Transcription factor AcademicSubjects/MED00305 Estradiol Cell growth Chemistry Estrogen Receptor alpha Estrogens Cell cycle single cell Cell biology GREB1 030104 developmental biology medicine.anatomical_structure Nuclear receptor 030220 oncology & carcinogenesis Carcinogens Estrogen receptor alpha |
| Zdroj: | Toxicological Sciences Toxicological Sciences, 181(2), 187-198. Oxford University Press (OUP) |
| ISSN: | 1096-0929 1096-6080 |
| DOI: | 10.1093/toxsci/kfab037 |
| Popis: | Estrogen receptor alpha (ERα) belongs to the nuclear hormone receptor family of ligand-inducible transcription factors and regulates gene networks in biological processes such as cell growth and proliferation. Disruption of these networks by chemical compounds with estrogenic activity can result in adverse outcomes such as unscheduled cell proliferation, ultimately culminating in tumor formation. To distinguish disruptive activation from normal physiological responses, it is essential to quantify relationships between different key events leading to a particular adverse outcome. For this purpose, we established fluorescent protein MCF7 reporter cell lines for ERα-induced proliferation by bacterial artificial chromosome-based tagging of 3 ERα target genes: GREB1, PGR, and TFF1. These target genes are inducible by the non-genotoxic carcinogen and ERα agonist 17β-estradiol in an ERα-dependent manner and are essential for ERα-dependent cell-cycle progression and proliferation. The 3 GFP reporter cell lines were characterized in detail and showed different activation dynamics upon exposure to 17β-estradiol. In addition, they demonstrated specific activation in response to other established reference estrogenic compounds of different potencies, with similar sensitivities as validated OECD test methods. This study shows that these fluorescent reporter cell lines can be used to monitor the spatial and temporal dynamics of ERα pathway activation at the single-cell level for more mechanistic insight, thereby allowing a detailed assessment of the potential carcinogenic activity of estrogenic compounds in humans. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|