BRAF and NRAS mutated melanoma: Different Ca2+ responses, Na+/Ca2+ exchanger expression, and sensitivity to inhibitors
BRAF and NRAS mutated melanoma: Different Ca2+ responses, Na+/Ca2+ exchanger expression, and sensitivity to inhibitors
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DOI：
10.1016/j.ceca.2020.102241
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发表时间：
2020-09-01
期刊：
影响因子：
4
通讯作者：
Rodrigues, Tiago
中科院分区：
文献类型：
--
作者：
Esteves, Gabriela Nohemi Nunez;Ferraz, Leticia Silva;Rodrigues, Tiago
钙离子是细胞内普遍存在的第二信使，在多种生物学过程的调节中起着重要作用。Ca 2+稳态和信号传导的改变是肿瘤细胞获得增殖和存活优势的重要特征，其包括储存能力、通道和泵的结构和功能变化。在这里，我们研究了维罗非尼反应性和非反应性黑色素瘤细胞中Ca 2+稳态的差异。此外，Na+/Ca 2+交换器（NCX）的表达及其抑制的影响进行了研究。为此，使用B-RAF（V600 E）和NRAS（Q61 R）突变的人黑素瘤细胞。细胞内Ca 2+螯合剂BAPTA-AM降低了SK-MEL-147的活力，但不降低SK-MEL-19和EGTA使NRAS（Q61 R）突变细胞对维罗非尼敏感的活力。与SK-MEL-19相比，这些细胞对thapsargin和ionomycin的细胞溶质Ca 2+水平的反应也较小，这与NCX 1表达增加、NO基础水平和对NCX抑制剂的敏感性有关。这些数据突出了B-RAF（V600 E）和NRAS（Q61 R）突变的黑色素瘤细胞对Ca 2+刺激的反应差异，并指出临床使用的化疗药物（包括维罗非尼）与NCX抑制剂的潜在组合作为治疗黑色素瘤的新治疗策略。
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.