Lu Chi-Cheng, Chen Hao-Ping, Chiang Jo-Hua, Jin Yi-An, Kuo Sheng-Chu, Wu Tian-Shung, Hour Mann-Jen, Yang Jai-Sing, Chiu Yu-Jen
Department of Food Science and Biotechnology, National Chung Hsing University, Taichung 402, Taiwan, R.O.C.
Department of Biochemistry, Tzu Chi University, Hualien 970, Taiwan, R.O.C.
Oncol Rep. 2014 Aug;32(2):597-606. doi: 10.3892/or.2014.3250. Epub 2014 Jun 12.
The aim of the present study was to explore the effect of 6-fluoro-2-(3-fluorophenyl)-4-(cyanoanilino) quinazoline (HMJ-30) on the anti-angiogenic properties and apoptosis-related mechanism of human umbilical vein endothelial cells (HUVECs). In this study, HMJ-30 dose- and time-dependently inhibited the viability of HUVECs. We also found that HMJ-30 enhanced disruption of tube-like structures and suppressed cell migration in HUVECs after vascular endothelial growth factor (VEGF) induction. HMJ-30 was also observed to inhibit vessel branching and sprouting in chicken chorioallantoic membrane (CAM). Microsprouting induced by VEGF in the rat aortic ring and blood vessel formation in a mouse Matrigel plug were individually suppressed by HMJ-30. In an in vitro study, HMJ-30 induced the apoptotic death of HUVECs as indicated by DNA fragmentation and promoted reactive oxygen species (ROS) production as determined by flow cytometric assay. In addition, extrinsic caspase signaling (caspase-8 and -3) was activated in the HMJ-30-treated HUVECs and their inhibitors were applied to assess the signal transduction. We investigated the upstream of the death receptor pathway and further observed that the levels of death receptor 5 (DR5) and phosphorylated c-Jun N-terminal kinase (JNK) signals were upregulated in HUVECs following HMJ-30 challenge, which was confirmed by a JNK-specific inhibitor (SP600125). Hence, HMJ-30-induced endothelial cell apoptosis involved the ROS/JNK-regulated DR5 pathway. In summary, HMJ-30 may provide a potential therapeutic effect for the anti-vascular targeting of angiogenesis during cancer treatment.
本研究旨在探讨6-氟-2-(3-氟苯基)-4-(氰基苯胺基)喹唑啉(HMJ-30)对人脐静脉内皮细胞(HUVECs)抗血管生成特性及凋亡相关机制的影响。在本研究中，HMJ-30呈剂量和时间依赖性抑制HUVECs的活力。我们还发现，HMJ-30可增强血管内皮生长因子(VEGF)诱导后HUVECs中管状结构的破坏并抑制细胞迁移。还观察到HMJ-30可抑制鸡胚绒毛尿囊膜(CAM)中的血管分支和出芽。HMJ-30分别抑制了VEGF在大鼠主动脉环中诱导的微出芽和小鼠基质胶栓中的血管形成。在一项体外研究中，HMJ-30诱导HUVECs发生凋亡性死亡，表现为DNA片段化，并通过流式细胞术检测发现其促进了活性氧(ROS)的产生。此外，在经HMJ-30处理的HUVECs中，外源性半胱天冬酶信号通路(半胱天冬酶-8和-3)被激活，并应用其抑制剂来评估信号转导。我们研究了死亡受体途径的上游，进一步观察到在HMJ-30刺激后，HUVECs中死亡受体5(DR5)和磷酸化c-Jun氨基末端激酶(JNK)信号的水平上调，这一结果得到了JNK特异性抑制剂(SP600125)的证实。因此，HMJ-30诱导的内皮细胞凋亡涉及ROS/JNK调节的DR5途径。总之，HMJ-30可能为癌症治疗期间抗血管生成的血管靶向治疗提供潜在的治疗效果。
