Epithelial cells that line the renal tubules possess apico-basal polarity and a defined cellular structure maintained by junctional proteins. The underlying mesenchymal cells are fibroblastic with a non-uniform cell structure and secrete extracellular matrix proteins. Mesenchymal to epithelial transition (MET) and epithelial to mesenchymal transition (EMT) are required for renal tubule formation during kidney development. Failure of renal mesenchymal cells to undergo epithelial transformation leads to the initiation of Wilms tumor, the most frequently occurring renal cancer in children. Conversely, aberrant mesenchymal transformation of tubular epithelial cells in the nephron contributes to the development of renal cell carcinoma, which constitutes 85-90% of the adult tubular malignancies.
The role of calcium regulators in governing EMT and MET is becoming evident. Sodium calcium exchanger 1 (NCX1), located on the basolateral surface of tubular epithelial cells, is the principal calcium regulator that mediates calcium reabsorption in these cells. NCX1 mediates the extrusion of one calcium ion and the influx of three sodium ions in one exchange movement. We demonstrated earlier that NCX1 regulates epithelial cell motility. However, the role of NCX1 in EMT was undetermined. We observed that knockdown of NCX1 in renal epithelial cells (MDCK) induced fibroblastic morphology, and increased permeability to rhodamine indicative of leaky cell-cell junctions. Electron micrographs of these cells displayed increased inter-cellular junctional distance also suggesting that NCX1 knockdown altered junctions between adjacent cells. Cells with NCX1 knockdown showed loss of apico-basal polarity in three-dimensional cultures accompanied by expression of mesenchymal markers. Furthermore, NCX1 knockdown cells were capable of anchorage independent growth suggesting that these cells had acquired tumorigenic potential. Interestingly, we show for the first time that NCX1 mRNA and protein expression is greatly reduced in both Wilms tumor and renal cell carcinoma demonstrating a direct correlation between NCX1 expression and the epithelial phenotype.
Mechanistically, we provide evidence that NCX1 interacts with and anchors E-cadherin, a classical adhesion molecule, to the cell surface independent of NCX1 ion transport activity. MDCK cells with NCX1 knockdown exhibited β-catenin nuclear localization and enhanced transcriptional activity. Taken together, knockdown of NCX1 in MDCK cells induces mesenchymal transition by destabilization of E-cadherin and induction of β-catenin transcriptional activity.
Citation Format: Sona Lakshme Balasubramaniam, Anilkumar Gopalakrishnapillai, Nicholas J. Petrelli, Sonali P. Barwe. Sodium-calcium exchanger-1 regulates the epithelial phenotype and is lost in renal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3931. doi:10.1158/1538-7445.AM2017-3931