BRAF and NRAS mutated melanoma: Different Ca2+ responses, Na+/Ca2+ exchanger expression, and sensitivity to inhibitors

Abstract

Calcium is a ubiquitous intracellular second messenger, playing central roles in the regulation of several biological processes. Alterations in Ca²⁺ homeostasis and signaling are an important feature of tumor cells to acquire proliferative and survival advantages, which include structural and functional changes in storage capacity, channels, and pumps. Here, we investigated the differences in Ca²⁺ homeostasis in vemurafenib-responsive and non-responsive melanoma cells. Also, the expression of the Na⁺/Ca²⁺ exchanger (NCX) and the impact of its inhibition were studied. For this, it was used B-RAF^V600E and NRAS^Q61R-mutated human melanoma cells. The intracellular Ca²⁺ chelator BAPTA-AM decreased the viability of SK-MEL-147 but not of SK-MEL-19 and EGTA sensitized NRAS^Q61R-mutated cells to vemurafenib. These cells also presented a smaller response to thapsargin and ionomycin regarding the cytosolic Ca²⁺ levels in relation to SK-MEL-19, which was associated to an increased expression of NCX1, NO basal levels, and sensitivity to NCX inhibitors. These data highlight the differences between B-RAF^V600E and NRAS^Q61R-mutated melanoma cells in response to Ca²⁺ stimuli and point to the potential combination of clinically used chemotherapeutic drugs, including vemurafenib, with NCX inhibitors as a new therapeutic strategy to the treatment of melanoma.

Keywords: Calcium; Cancer; Cell death; Na(+)/Ca(2+)exchanger; Targeted therapy.