Istituto Dermopatico dell'Immacolata, IDI-IRCCS, 00167 Rome, Italy.
Int J Mol Sci. 2024 Mar 22;25(7):3589. doi: 10.3390/ijms25073589.
Miconazole is an antimycotic drug showing anti-cancer effects in several cancers. However, little is known on its effects in melanoma. A375 and SK-MEL-28 human melanoma cell lines were exposed to miconazole and clotrimazole (up to 100 mM). Proliferation, viability with MTT assay and vascular mimicry were assayed at 24 h treatment. Molecular effects were measured at 6 h, namely, ATP-, ROS-release and mitochondria-related cytofluorescence. A metabolomic profile was also investigated at 6 h treatment. Carnitine was one of the most affected metabolites; therefore, the expression of 29 genes involved in carnitine metabolism was investigated in the public platform GEPIA2 on 461 melanoma patients and 558 controls. After 24 h treatments, miconazole and clotrimazole strongly and significantly inhibited proliferation in the presence of 10% serum on either melanoma cell lines; they also strongly reduced viability and vascular mimicry. After 6 h treatment, ATP reduction and ROS increase were observed, as well as a significant reduction in mitochondria-related fluorescence. Further, in A375, miconazole strongly and significantly altered expression of several metabolites including carnitines, phosphatidyl-cholines, all amino acids and several other small molecules, mostly metabolized in mitochondria. The expression of 12 genes involved in carnitine metabolism was found significantly modified in melanoma patients, 6 showing a significant impact on patients' survival. Finally, miconazole antiproliferation activity on A375 was found completely abrogated in the presence of carnitine, supporting a specific role of carnitine in melanoma protection toward miconazole effect, and was significantly reversed in the presence of caspases inhibitors such as ZVAD-FMK and Ac-DEVD-CHO, and a clear pro-apoptotic effect was observed in miconazole-treated cells, by FACS analysis of Annexin V-FITC stained cells. Miconazole strongly affects proliferation and other biological features in two human melanoma cell lines, as well as mitochondria-related functions such as ATP- and ROS-release, and the expression of several metabolites is largely dependent on mitochondria function. Miconazole, likely acting via carnitine and mitochondria-dependent apoptosis, is therefore suggested as a candidate for further investigations in melanoma treatments.
咪康唑是一种抗真菌药物，在几种癌症中显示出抗癌作用。然而，关于其在黑色素瘤中的作用知之甚少。将 A375 和 SK-MEL-28 人黑色素瘤细胞系暴露于咪康唑和克霉唑（高达 100mM）。在 24 小时处理后，通过 MTT 测定法测定增殖、活力和血管模拟。在 6 小时时测量分子效应，即 ATP-、ROS 释放和线粒体相关的细胞荧光。还在 6 小时处理时研究了代谢组学图谱。肉碱是受影响最大的代谢物之一；因此，在 461 名黑色素瘤患者和 558 名对照者的公共平台 GEPIA2 上调查了 29 个与肉碱代谢相关的基因的表达。经过 24 小时的治疗，咪康唑和克霉唑在存在 10%血清的情况下强烈且显著地抑制了两种黑色素瘤细胞系的增殖；它们还强烈降低了活力和血管模拟。6 小时后，观察到 ATP 减少和 ROS 增加，以及线粒体相关荧光的显著减少。此外，在 A375 中，咪康唑强烈且显著地改变了几种代谢物的表达，包括肉碱、磷脂酰胆碱、所有氨基酸和几种其他小分子，这些代谢物主要在线粒体中代谢。在黑色素瘤患者中发现 12 个与肉碱代谢相关的基因的表达明显改变，其中 6 个对患者的生存有显著影响。最后，在存在肉碱的情况下，咪康唑对 A375 的增殖活性完全被阻断，这支持了肉碱在保护黑色素瘤免受咪康唑作用方面的特定作用，并且在存在半胱天冬酶抑制剂如 ZVAD-FMK 和 Ac-DEVD-CHO 时被显著逆转，并且在咪康唑处理的细胞中通过 Annexin V-FITC 染色细胞的 FACS 分析观察到明显的促凋亡作用。咪康唑强烈影响两种人黑色素瘤细胞系的增殖和其他生物学特征，以及与线粒体相关的功能，如 ATP-和 ROS 释放，并且几种代谢物的表达在很大程度上依赖于线粒体功能。因此，咪康唑可能通过肉碱和线粒体依赖性细胞凋亡起作用，因此被提议作为黑色素瘤治疗进一步研究的候选药物。
