Department of Beauty Science, Meiho University, Pingtung 91202, Taiwan.
Department of Food and Nutrition, Meiho University, Pingtung 91202, Taiwan.
Int J Mol Sci. 2020 May 25;21(10):3737. doi: 10.3390/ijms21103737.
(+)-Bornyl -coumarate is an active substance that is abundant in the stem and has been shown to possess bioactivity against bacteria and a strong antioxidative effect. In the current study, we examined the actions of (+)-bornyl -coumarate against A2058 and A375 melanoma cells. The inhibition effects of (+)-bornyl -coumarate on these cell lines were assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay and the underlying mechanisms were identified by immunostaining, flow cytometry and western blotting of proteins associated with apoptosis and autophagy. Our results demonstrated that (+)-bornyl -coumarate inhibited melanoma cell proliferation and caused loss of mitochondrial membrane potential, demonstrating treatment induced apoptosis. In addition, western blotting revealed that the process is mediated by caspase-dependent pathways, release of cytochrome , activation of pro-apoptotic proteins (Bax, Bad and caspase-3/-9) and suppression of anti-apoptotic proteins (Bcl-2, Bcl-xl and Mcl-1). Also, the upregulated expressions of -PERK, p-eIF2α, ATF4 and CCAAT/enhancer-binding protein (C/EBP)-homologous protein (CHOP) after treatment indicated that (+)-bornyl -coumarate caused apoptosis via endoplasmic reticulum (ER) stress. Moreover, increased expressions of beclin-1, Atg3, Atg5, p62, LC3-I and LC3-II proteins and suppression by autophagic inhibitor 3-methyladenine (3-MA), indicated that (+)-bornyl -coumarate triggered autophagy in the melanoma cells. In conclusion, our findings demonstrated that (+)-bornyl -coumarate suppressed human melanoma cell growth and should be further investigated with regards to its potential use as a chemotherapy drug for the treatment of human melanoma.
（+）-莰烯 - 香豆素是一种丰富存在于茎中的活性物质，已被证明具有抗细菌活性和强大的抗氧化作用。在本研究中，我们研究了（+）-莰烯 - 香豆素对 A2058 和 A375 黑色素瘤细胞的作用。通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴化物（MTT）测定评估（+）-莰烯 - 香豆素对这些细胞系的抑制作用，并通过免疫染色、流式细胞术和与凋亡和自噬相关的蛋白质的蛋白质印迹鉴定潜在机制。我们的结果表明，（+）-莰烯 - 香豆素抑制黑色素瘤细胞增殖并导致线粒体膜电位丧失，表明治疗诱导凋亡。此外，蛋白质印迹显示该过程是通过 caspase 依赖性途径介导的，释放细胞色素 C，激活促凋亡蛋白（Bax、Bad 和 caspase-3/-9）并抑制抗凋亡蛋白（Bcl-2、Bcl-xl 和 Mcl-1）。此外，处理后 -PERK、p-eIF2α、ATF4 和 CCAAT/增强子结合蛋白（C/EBP）同源蛋白（CHOP）的上调表达表明（+）-莰烯 - 香豆素通过内质网（ER）应激引起细胞凋亡。此外，自噬抑制剂 3-甲基腺嘌呤（3-MA）抑制 beclin-1、Atg3、Atg5、p62、LC3-I 和 LC3-II 蛋白的表达增加，表明（+）-莰烯 - 香豆素在黑色素瘤细胞中引发自噬。总之，我们的研究结果表明，（+）-莰烯 - 香豆素抑制人黑色素瘤细胞生长，应进一步研究其作为治疗人类黑色素瘤的化疗药物的潜力。
