Context: Therapy of thyroid carcinoma utilizes its radioiodine concentration ability for treatment. Dedifferentiated cells lose radioiodine uptake from sodium-iodide symporter (hNIS) gene transcription failure consequent to genomic structure (chromatin compaction) and composition (CpG methylation).

Objective/Methods: We explored restoring hNIS expression in human thyroid carcinoma cells using thyroid adenoma and carcinoma cell lines: KAK-1, NPA'87, BHT-101, and KAT-4B, with quantitative RT-PCR, chromatin immunoprecipitation, DNase I sensitivity assays, and luciferase reporter construct transfections containing hNIS promoter regions.

Results: Combined 5-azacytidine (azaC) and sodium butyrate (NaB) restores hNIS gene transcription in KAK-1 to levels approaching radioiodine-treatable tumors. Despite induction of H4 acetylation, there was no DNase I sensitivity enhancement in two regions of the hNIS gene promoter. Cycloheximide (CHX) in cells transfected with luciferase reporter construct, 1.3 kb hNIS gene promoter, stimulated normalized luciferase expression, singly and synergistically with azaC, in dose-dependent, time course-dependent, cell type-specific, and promoter-specific fashion. Both anisomycin and emetine, but not puromycin, had similar effects. CHX also increased endogenous hNIS mRNA. Transfections with reporter constructs containing consecutive deletions of hNIS gene promoter sequences revealed responsible sequences at -427 to -131 bp. Deletion of 1.2 kb promoter region upstream of -131 bp enhanced basal luciferase reporter activity 3-fold above the activity of full length promoter construct, supporting inhibitory properties of this region.

Conclusions: This suggests that trans-active protein factor(s) represses endogenous hNIS transcription in KAK-1 cells under basal conditions, accounting for loss of iodine uptake. Inhibition of this repressive activity increases endogenous hNIS transcription and presents a novel target to restore hNIS expression in dedifferentiated thyroid carcinoma.