NADPH Oxidase NOX4 Is a Critical Mediator of BRAFV600E-Induced Downregulation of the Sodium/Iodide Symporter in Papillary Thyroid Carcinomas

Aims: The BRAF(V600E) oncogene, reported in 40%-60% of papillary thyroid cancer (PTC), has an important role in the pathogenesis of PTC. It is associated with the loss of thyroid iodide-metabolizing genes, such as sodium/iodide symporter (NIS), and therefore with radioiodine refractoriness. Inhibition of mitogen-activated protein kinase (MAPK) pathway, constitutively activated by BRAF(V600E), is not always efficient in resistant tumors suggesting that other compensatory mechanisms contribute to a BRAF(V600E) adaptive resistance. Recent studies pointed to a key role of transforming growth factor beta (TGF-beta) in BRAF(V600E) -induced effects. The reactive oxygen species (ROS)-generating NADPH oxidase NOX4, which is increased in PTC, has been identified as a new key effector of TGF-b in cancer, suggestive of a potential role in BRAF(V600E) -induced thyroid tumors. Results: Here, using two human BRAF(V600E) -mutated thyroid cell lines and a rat thyroid cell line expressing BRAF(V600E) in a conditional manner, we show that NOX4 upregulation is controlled at the transcriptional level by the oncogene via the TGF-beta/Smad3 signaling pathway. Importantly, treatment of cells with NOX4-targeted siRNA downregulates BRAF(V600E) -induced NIS repression. Innovation and Conclusion: Our results establish a link between BRAF(V600E) and NOX4, which is confirmed by a comparative analysis of NOX4 expression in human (TCGA) and mouse thyroid cancers. Remarkably, analysis of human and murine BRAF(V600E) -mutated thyroid tumors highlights that the level of NOX4 expression is inversely correlated to thyroid differentiation suggesting that other genes involved in thyroid differentiation in addition to NIS might be silenced by a mechanism controlled by NOX4-derived ROS. This study opens a new opportunity to optimize thyroid cancer therapy. Antioxid. Redox Signal. 26, 864-877.