A new bioluminescent cellular assay to measure the transcriptional effects of chemicals that modulate the alpha-1 thyroid hormone receptor.

摘要: Abstract Interactions of environmental pollutants with the thyroid endocrine axis have received much attention especially because hormones (THs) play a major role in mammalian brain development. In order to screen for compounds that act on triiodothyronine (T3) signaling pathway, we developed new reporter gene assay expressing luciferase under control TH receptor (TR). PC12 cells α1-isoform TR avian origin were stably transfected controlled by SV40 promoter, and enhanced four-spaced direct repeat (DR4) response element (TRE). The resulting PC-DR-LUC used optimize T3 multiwell microplates. This was highly sensitive (30 pM T3) reproducible, responded as expected analogues. Several halogenated phenolic (3,3′,5,5′-tetrabromobisphenol A, 3,3′,5,5′-tetrachlorobisphenol 4-hydroxy-2′,3,4′,5,6′-pentachlorobiphenyl) phenol (pentachlorophenol, 2,4,6-triiodophenol) suspected being thyroid-disrupting chemicals induced partial agonistic and/or complex competitive/uncompetitive antagonistic responses at micromolar concentrations. A cell viability test indicated these effects not related cytotoxicity chemicals. These results suggest could be valuable tool large-scale screening agonists antagonists vitro , detecting disruptors environment.