Imatinib mesylate induction of ROS-dependent apoptosis in melanoma B16F0 cells
Imatinib mesylate induction of ROS-dependent apoptosis in melanoma B16F0 cells
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DOI：
10.1016/j.jdermsci.2011.03.001
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发表时间：
2011-06-01
影响因子：
4.6
通讯作者：
Chen, Yen-Chou
中科院分区：
文献类型：
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作者：
Chang, Shao-Ping;Shen, Shing-Chuan;Chen, Yen-Chou
背景：蛋白酪氨酸激酶抑制剂甲磺酸伊马替尼(STI571)可通过诱导细胞凋亡降低肿瘤细胞的存活率；目的：本研究探讨了ROS在STI571诱导黑色素瘤B16F0细胞凋亡中的作用，阐明了STI571诱导黑色素瘤B16F0细胞凋亡的机制。方法：采用体外细胞培养系统，通过Western blotting、流式细胞仪分析、酶活性测定和DNA完整性分析等方法检测STI571对细胞内ROS生成、细胞周期进程、半胱氨酸酶活性和线粒体功能的影响。在药理学研究中，应用ROS清除剂N-乙酰半胱氨酸(NAC)、NADPH氧化酶抑制剂二苯碘(DPI)和丝裂原活化蛋白激酶(MAPK)抑制剂(PD98059、SP600125和SB203580)来研究其作用机制。结果：STI571通过诱导细胞凋亡降低了黑色素瘤细胞B16F0的活性，但不影响人皮肤成纤维细胞WS1的活性。此外，加入NAC和DPI可抑制STI571诱导的细胞凋亡，并可使黑色素瘤B16F0细胞内过氧化脂质水平升高。在STI571处理的B16F0细胞中，caspase3和9酶活性被激活，同时线粒体膜电位被破坏，这与刺激JNK和p38蛋白磷酸化相一致。首次在黑色素瘤B16F0细胞中发现STI571介导的ROS依赖的细胞凋亡，并被JNK抑制剂SP600125增强。结论：我们的结果支持STI571诱导黑色素瘤B16F0细胞ROS依赖的细胞凋亡的观点。建议将JNK抑制剂与STI571联合用于治疗黑色素瘤，以进行进一步的体内研究。(C)2011年日本皮肤病研究学会。爱思唯尔爱尔兰有限公司出版。版权所有。
Background: Imatinib mesylate (STI571), a protein tyrosine kinase inhibitor, was shown to reduce the viability of several cancer cell lines via apoptosis induction; however, the role of reactive oxygen species (ROS) in STI571-induced melanoma cell apoptosis is still undefined.Objective: In this study, we investigated the contribution of ROS to STI571-induced apoptosis in melanoma B16F0 cells, and the apoptotic mechanism elicited by STI571 was illustrated.Methods: Using an in vitro cell culture system, the effects of STI571 on ROS production, cell cycle progression, caspase activation, and mitochondrial functions were examined via Western blotting, a flow cytometric analysis, an enzyme activity assay, and a DNA integrity assay. In pharmacological studies, the ROS scavenger, N-acetyl cysteine (NAC), the NADPH oxidase inhibitor, dipheylene iodide (DPI), and mitogen-activated protein kinase (MAPK) inhibitors (PD98059, SP600125, and SB203580) were applied to investigate the mechanism.Results: STI571 reduced the viability of melanoma cells B16F0, but not human skin fibroblasts WS1, via apoptosis induction. Besides, apoptosis induced by STI571 was inhibited by the addition of NAC and DPI, and an increase in the intracellular peroxide level by STI571 was identified in melanoma B16F0 cells. Activation of caspases 3 and 9 enzyme activities accompanied by disrupting the mitochondria membrane potential in according with stimulating JNK and p38 protein phosphorylation was identified in STI571-treated B16F0 cells. STI571-mediated a ROS-dependent apoptosis potentiated by JNK inhibitor SP600125 was first identified in melanoma B16F0 cells.Conclusion: Our results support the idea that ROS-dependent apoptosis in STI571-treated melanoma cells B16F0. The combination of a JNK inhibitor with STI571 for treating melanomas is suggested for further in vivo studies. (C) 2011 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.