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Toxicol Sci. 2019 Jan 10. doi: 10.1093/toxsci/kfz004. [Epub ahead of print]

Improving Estrogenic Compound Screening Efficiency by Using Self-Modulating, Continuously Bioluminescent Human Cell Bioreporters Expressing a Synthetic Luciferase.

Author information

1
Center For Environmental Biotechnology, University of Tennessee, Knoxville, TN, USA.
2
490 BioTech, Inc., Knoxville TN, USA.

Abstract

A synthetic bacterial luciferase-based autobioluminescent bioreporter, HEK293ERE/Gal4-Lux, was developed in a human embryonic kidney (HEK293) cell line for the surveillance of chemicals displaying endocrine disrupting activity. Unlike alternative luminescent reporters, this bioreporter generates bioluminescence autonomously without requiring an external light-activating chemical substrate or cellular destruction. The bioreporter's performance was validated against a library of 76 agonistic and antagonistic estrogenic endocrine disruptor chemicals and demonstrated reproducible half maximal effective concentration (EC50) values meeting the U.S. Environmental Protection Agency (EPA) guidelines for Tier 1 endocrine disrupting chemical screening assays. For model compounds, such as the estrogen receptor (ER) agonist 17β-estradiol, HEK293ERE/Gal4-Lux demonstrated an EC50 value (7.9 × 10-12 M) comparable to that of the current EPA-approved HeLa-9903 firefly luciferase-based estrogen receptor transcription assay (4.6 × 10-12 M). Screening against an expanded array of common ER agonists likewise produced similar relative effect potencies as compared to existing assays. The self-initiated autobioluminescent signal of the bioreporter permitted facile monitoring of the effects of endocrine disrupting chemicals, which decreased the cost and hands-on time required to perform these assays. These characteristics make the HEK293ERE/Gal4-Lux bioreporter potentially suitable as a high-throughput human cell-based assay for screening estrogenic activity.

PMID:
30629247
DOI:
10.1093/toxsci/kfz004

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