DC-81-enediyne induces apoptosis of human melanoma A375 cells: involvement of the ROS, p38 MAPK, and AP-1 signaling pathways
DC-81-enediyne induces apoptosis of human melanoma A375 cells: involvement of the ROS, p38 MAPK, and AP-1 signaling pathways
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DOI：
10.1007/s10565-012-9238-6
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发表时间：
2013-04-01
影响因子：
6.1
通讯作者：
Hu, Wan-Ping
中科院分区：
文献类型：
--
作者：
Chen, Chung-Yu;Chen, Yin-Kai;Hu, Wan-Ping
黑色素瘤是患者护理中最耐药的癌症之一。目前治疗的缓解率仍然很低。 DC-81是一种由链霉菌属产生的抗肿瘤抗生素，属于吡咯并[2,1-c][1,4]苯二氮卓类（PBD），是一种有效的核酸合成抑制剂。烯二炔含有 DNA 嵌入基团或 DNA 小沟结合功能，由于它们能够产生苯环双自由基，因此它们是有效的 DNA 损伤剂。我们之前报道了一系列与烯二炔连接的新型 PBD 杂化物的有效合成和抗肿瘤活性。本研究的目的是探讨DC-81-烯二炔剂对人黑色素瘤A375细胞的抗增殖作用的机制。通过流式细胞术检测发现，DC-81-烯二炔诱导 Ca2+ 水平和活性氧 (ROS) 生成增加。蛋白质印迹分析表明，DC-81-烯二炔诱导 p38 磷酸化和激活转录因子 2 (ATF-2)。通过使用荧光素酶报告基因测定，用分级浓度的 DC-81-烯二炔处理 A375 细胞后，激活蛋白 1 (AP-1) 活性进一步增强。添加 Ca2+、ROS 和 p38 抑制剂可显着消除 DC-81-烯二炔处理诱导的 A375 细胞凋亡。总的来说，我们的研究表明，DC-81-烯二炔通过增加 Ca2+ 和 ROS 生成诱导 A375 细胞凋亡，其中涉及 p38 磷酸化和增强 ATF-2/AP-1 表达，导致 caspase-3 活性、聚 (ADP-核糖) 聚合酶裂解、M30 CytoDeath 染色和随后的细胞凋亡。
Melanoma is one of the most chemoresistant cancers in patient care. The remission rate of current therapy remains low. DC-81, an antitumor antibiotic produced by Streptomyces species, belongs to pyrrolo[2,1-c][1,4]benzodiazepine (PBD), which is a potent inhibitor of nucleic acid synthesis. An enediyne contains either DNA intercalating groups or DNA minor groove binding functions and these are potent DNA-damaging agents due to their ability to generate benzenoid diradicals. We have previously reported an efficient synthesis and antitumor activity of a series of novel PBD hybrids linked with enediyne. The purpose of this study was to examine the mechanism of the antiproliferative effect of DC-81-enediyne agent on human melanoma A375 cells. DC-81-enediyne induced an increase in Ca2+ level and reactive oxygen species (ROS) generation as detected by flow cytometric assay. Western blot analysis showed that DC-81-enediyne induced the phosphorylation of p38 and activating transcription factor 2 (ATF-2). By using the luciferase reporter assay, activating protein-1 (AP-1) activity was further enhanced after A375 cells were treated with graded concentrations of DC-81-enediyne. DC-81-enediyne treatment-induced A375 cell apoptosis was significantly abrogated by the addition of Ca2+, ROS, and p38 inhibitors. Collectively, our studies indicate that DC-81-enediyne induces A375 cell apoptosis through an increased Ca2+ and ROS generation, which involves p38 phosphorylation and enhanced ATF-2/AP-1 expressions, leading to caspase-3 activity, poly(ADP-ribose)polymerase cleavage, M30 CytoDeath staining, and subsequent apoptotic cell death.