PDF(Pubmed)
Abstract：
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.
摘要：
咪康唑是一种在几种癌症中显示抗癌作用的抗真菌药物。然而,对黑色素瘤的影响知之甚少。将A375和SK-MEL-28人黑素瘤细胞系暴露于咪康唑和克霉唑(高达100mM)。扩散,在24小时处理时，用MTT测定法测定活力和血管拟态。在6小时测量分子效应，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概况。肉碱是受影响最大的代谢物之一；因此，我们在公共平台GEPIA2中对461例黑色素瘤患者和558例对照中29个参与肉碱代谢的基因的表达进行了调查.24小时治疗后，在存在10％血清的情况下，咪康唑和克霉唑强烈且显着地抑制了两种黑色素瘤细胞系的增殖；它们还大大降低了活力和血管拟态。治疗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着减少。Further,在A375中，咪康唑强烈且显著改变了几种代谢物的表达，包括肉碱，磷脂酰胆碱,所有的氨基酸和其他一些小分子，主要在线粒体代谢。在黑色素瘤患者中发现12个参与肉碱代谢的基因表达显著改变，图6显示了对患者生存的显著影响。最后,发现在存在肉碱的情况下，咪康唑对A375的抗增殖活性完全消除，支持肉碱在黑色素瘤保护中对咪康唑作用的特定作用，并且在存在caspases抑制剂如ZVAD-FMK和Ac-DEVD-CHO时显著逆转，在咪康唑处理的细胞中观察到明显的促凋亡作用，通过膜联蛋白V-FITC染色细胞的FACS分析。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，以及线粒体相关功能，如ATP和ROS释放，几种代谢物的表达在很大程度上依赖于线粒体功能。咪康唑,可能通过肉碱和线粒体依赖性凋亡起作用，因此，建议作为黑色素瘤治疗进一步研究的候选人。
