Development of a non-radioactive screening assay to detect chemicals disrupting the human sodium iodide symporter activity.

Affiliation

Environmental Health Science and Research Bureau, Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, ON K1A 0K9, Canada. Electronic address: [Email]

Abstract

Adequate concentration of iodide ions within thyroid epithelial cells, which is mediated by the sodium iodide symporter (NIS), is essential for proper thyroid hormone synthesis. Inhibition of NIS activity represents a potential mechanism by which goitrogens/toxicants can disrupt thyroid hormone physiology. It is necessary to develop a rapid, simple, inexpensive and sensitive screening assay to identify chemicals affecting NIS function. The current study compares the sensitivities of non-radioactive Sandell-Kolthoff (SK) reaction and radioactive iodide uptake (RAIU) in a previously described NIS assay. The EPAhNIS cell line (HEK293T stably transfected to over-express the human NIS) was tested with the reference NIS inhibitor (sodium perchlorate) across multiple log concentration range. The results from SK reaction in EPAhNIS cells showed similar performance to published RAIU results from the same cell line, in terms of assay screening coefficient (Z') and variability (CV). Results from the reference chemicals tested in EPAhNIS cells revealed that SK reaction yielded IC50 and selectivity scores consistent with those observed for RAIU. However, RAIU seems marginally more sensitive than the SK reaction, as RAIU consistently detected weaker NIS inhibitors among the test chemicals. We developed a second hNIS assay based on the MCF-7 cell line. Applying reference anions and chemicals to MCF7hNIS cells, we found that in comparison with results from EPAhNIS cells, the SK reaction with MCF7hNIS: 1) yielded similar Z' and CV; 2) had similar IC50 and selectivity scores for reference chemicals; 3) identified more NIS inhibitors among reference chemicals than SK reaction, but less than the RAIU assay in EPAhNIS cells. In conclusion, the SK reaction can be used with both EPAhNIS and MCF7hNIS cells to measure iodide uptake and identify NIS inhibitors, except for those presenting an extremely weak potency.

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