* Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
† Department of Biochemistry and Molecular Cell Biology, College of Medicine, Taipei Medical University, Taipei, Taiwan.
Am J Chin Med. 2017;45(4):879-899. doi: 10.1142/S0192415X17500471. Epub 2017 May 18.
Evodiamine (EVO) is an active medicinal compound derived from the traditional herbal medicine Evodia rutaecarpa. It has been reported that evodiamine has several beneficial biological properties, including anticancer and anti-inflammatory activities. However, the in vitro and in vivo anticancer activities of EVO against the growth of glioblastoma cells remain undefined. EVO induced significant decreases in the viability of U87 and C6 glioma cells, but not of primary astrocytes, according with the occurrence of apoptotic characteristics including DNA ladders, caspase-3 and poly(ADP ribose) polymerase (PARP) protein cleavage, and hypodiploid cells. The disruption of the mitochondrial membrane potential (MMP) was detected, and it was found that the peptidyl caspase-9 inhibitor, Z-LEHD-FMK, significantly prevented glioma cells from EVO-induced apoptosis. Increased c-Jun N-terminal kinase (JNK) protein phosphorylation by EVO was observed, and the addition of JNK inhibitors, SP600125 and JNKI inhibited the EVO-induced apoptosis was inhibited. Additionally, EVO treatment induced G2/M arrest with increased polymerized tubulin protein expression in U87 and C6 cells. Elevated expressions of the cyclin B1, p53, and phosphorylated (p)-p53 proteins were detected in EVO-treated glioma cells, and these were inhibited by JNK inhibitors. An in vivo study showed that EVO significantly reduced the growth of gliomas elicited by the subcutaneous injection of U87 cells with increases in cyclin B1, p53, and p-p53 protein expressions in tumors. An analysis of eight EVO-related chemicals showed that alkyl groups at position 14 in EVO are important for its anti-glioma effects which involve both apoptosis and G2/M arrest. Evidence is provided that supports EVO induction of apoptosis and G2/M arrest via the activation of JNK-mediated gene expression and disruption of MMP in glioblastoma cells. EVO was shown to penetrate the blood-brain barrier; EVO is therefore predicted to be a promising compound for the chemotherapy of glioblastomas and deserves further investigations.
吴茱萸碱（EVO）是一种从传统草药吴茱萸中提取的活性药用化合物。据报道，吴茱萸碱具有多种有益的生物学特性，包括抗癌和抗炎活性。然而，EVO 对神经胶质瘤细胞生长的体外和体内抗癌活性仍未确定。EVO 显著降低 U87 和 C6 神经胶质瘤细胞的活力，但对原代星形胶质细胞没有影响，同时出现了凋亡特征，包括 DNA 梯状带、半胱天冬酶-3 和多聚（ADP 核糖）聚合酶（PARP）蛋白裂解以及亚二倍体细胞。检测到线粒体膜电位（MMP）的破坏，并且发现肽基半胱天冬酶-9 抑制剂 Z-LEHD-FMK 可显著阻止神经胶质瘤细胞发生 EVO 诱导的凋亡。EVO 引起 c-Jun N-末端激酶（JNK）蛋白磷酸化增加，并且添加 JNK 抑制剂 SP600125 和 JNKI 可抑制 EVO 诱导的凋亡。此外，EVO 处理诱导 U87 和 C6 细胞中微管蛋白聚合蛋白表达增加导致 G2/M 期阻滞。EVO 处理的神经胶质瘤细胞中检测到细胞周期蛋白 B1、p53 和磷酸化（p）-p53 蛋白的表达升高，这些表达被 JNK 抑制剂抑制。体内研究表明，EVO 可显著减少皮下注射 U87 细胞引起的神经胶质瘤的生长，并增加肿瘤中细胞周期蛋白 B1、p53 和 p-p53 蛋白的表达。对 8 种 EVO 相关化学物质的分析表明，EVO 中 14 位的烷基基团对于其抗癌作用很重要，这涉及到凋亡和 G2/M 期阻滞。研究结果提供了证据表明，EVO 通过激活 JNK 介导的基因表达和破坏 MMP 诱导神经胶质瘤细胞凋亡和 G2/M 期阻滞。EVO 被证明可以穿透血脑屏障；因此，EVO 有望成为治疗神经胶质瘤的有前途的化合物，值得进一步研究。
