Development and Characterization of a Human Reporter Cell Line for the Assessment of Thyroid Receptor Transcriptional Activity: A Case of Organotin Endocrine Disruptors

• Published in: Journal of agricultural and food chemistry 2015-08, Vol.63 (31), p.7074-7083
• Main Authors: Illés, Peter, Brtko, Július, Dvořák, Zdeněk
• Format: Article
• Language: English
• Subjects: cell biology; Cell Line - drug effects; Cell Line - metabolism; cryopreservation; dose response; Endocrine Disruptors - pharmacology; endocrine-disrupting chemicals; gene expression regulation; Genes, Reporter - drug effects; Humans; ligands; luciferase; Promoter Regions, Genetic - drug effects; Receptors, Thyroid Hormone - genetics; Receptors, Thyroid Hormone - metabolism; retinoic acid; steroid hormones; tin; transcription (genetics); Transcriptional Activation - drug effects; Transfection; tributyltin; triiodothyronine; Triiodothyronine - metabolism
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/acs.jafc.5b01519

We developed and characterized the human luciferase reporter cell line PZ-TR for the assessment of thyroid receptor (TR) transcriptional activity. Triiodothyronine (T3) induced luciferase activity in a dose-dependent manner, and the sensitivity of assay allowed for the detection of nanomolar T3 concentrations. The luciferase activity was induced by a maximum of (2.42 ± 0.14)–(2.73 ± 0.23)-fold after 24 h of exposure to 10 nM T3. We did not observe a nonspecific induction of luciferase activity by other steroid hormones and VDR ligands, with the exception of partial activation by retinoic acids. Cryopreservation of PZ-TR cells did not influence their functionality, responsivity to T3, or cell morphology, even after long-term cultivation. PZ-TR cells were used to evaluate the effects of organic tin compounds on TR. We found that the tributyltin and triphenyltin derivatives induced luciferase activity and that application of organotins in combination with T3 enhanced the effect of T3. These findings indicate that organic tin compounds have potential to interfere with TR-mediated regulation of gene expression and influence the physiological activity of thyroid hormones.