National Museum of Marine Biology and Aquarium, Pingtung 94450, Taiwan.
Mar Drugs. 2013 Jul 22;11(7):2625-42. doi: 10.3390/md11072625.
Sinulariolide is an active compound isolated from the cultured soft coral Sinularia flexibilis. In this study, we investigated the effects of sinulariolide on A375 melanoma cell growth and protein expression. Sinulariolide suppressed the proliferation and migration of melanoma cells in a concentration-dependent manner and was found to induce both early and late apoptosis by flow cytometric analysis. Comparative proteomic analysis was conducted to investigate the effects of sinulariolide at the molecular level by comparison between the protein profiles of melanoma cells treated with sinulariolide and those without treatment. Two-dimensional gel electrophoresis (2-DE) master maps of control and treated A375 cells were generated by analysis with PDQuest software. Comparison between these maps showed up- and downregulation of 21 proteins, seven of which were upregulated and 14 were downregulated. The proteomics studies described here identify some proteins that are involved in mitochondrial dysfunction and apoptosis-associated proteins, including heat shock protein 60, heat shock protein beta-1, ubiquinol cytochrome c reductase complex core protein 1, isocitrate dehydrogenase (NAD) subunit alpha (down-regulated), and prohibitin (up-regulated), in A375 melanoma cells exposed to sinulariolide. Sinulariolide-induced apoptosis is relevant to mitochondrial-mediated apoptosis via caspase-dependent pathways, elucidated by the loss of mitochondrial membrane potential, release of cytochrome c, and activation of Bax, Bad and caspase-3/-9, as well as suppression of p-Bad, Bcl-xL and Bcl-2. Taken together, our results show that sinulariolide-induced apoptosis might be related to activation of the caspase cascade and mitochondria dysfunction pathways. Our results suggest that sinulariolide merits further evaluation as a chemotherapeutic agent for human melanoma.
辛拉瑞内酯是从软珊瑚 Sinularia flexibilis 培养物中分离得到的一种活性化合物。在本研究中，我们研究了辛拉瑞内酯对 A375 黑色素瘤细胞生长和蛋白表达的影响。辛拉瑞内酯以浓度依赖的方式抑制黑色素瘤细胞的增殖和迁移，并通过流式细胞术分析发现诱导早期和晚期凋亡。通过比较用辛拉瑞内酯处理和未处理的黑色素瘤细胞的蛋白质谱，进行比较蛋白质组学分析以从分子水平研究辛拉瑞内酯的作用。用 PDQuest 软件分析生成对照和处理的 A375 细胞的二维凝胶电泳 (2-DE) 图谱。通过比较这些图谱，显示 21 种蛋白质上调和下调，其中 7 种上调，14 种下调。这里描述的蛋白质组学研究鉴定了一些参与线粒体功能障碍和凋亡相关蛋白的蛋白质，包括热休克蛋白 60、热休克蛋白 beta-1、泛醌细胞色素 c 还原酶核心蛋白 1、异柠檬酸脱氢酶（NAD）亚基 alpha（下调）和抑制素（上调），在暴露于辛拉瑞内酯的 A375 黑色素瘤细胞中。辛拉瑞内酯诱导的细胞凋亡与通过 caspase 依赖性途径的线粒体介导的细胞凋亡有关，这通过线粒体膜电位丧失、细胞色素 c 释放以及 Bax、Bad 和 caspase-3/-9 的激活以及 p-Bad、Bcl-xL 和 Bcl-2 的抑制来阐明。总之，我们的结果表明，辛拉瑞内酯诱导的细胞凋亡可能与 caspase 级联和线粒体功能障碍途径的激活有关。我们的结果表明，辛拉瑞内酯作为人类黑色素瘤的化疗剂值得进一步评估。
