Evodiamine from Evodia rutaecarpa induces apoptosis via activation of JNK and PERK in human ovarian cancer cells
Evodiamine from Evodia rutaecarpa induces apoptosis via activation of JNK and PERK in human ovarian cancer cells
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DOI：
10.1016/j.phymed.2015.12.003
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发表时间：
2016-01-15
期刊：
影响因子：
7.9
通讯作者：
Chen, Yen-Chou
中科院分区：
文献类型：
--
作者：
Chen, Tze-Chien;Chien, Chih-Chiang;Chen, Yen-Chou
背景：吴茱萸二胺(Evodiine，EVO；8，13，13b，14-tetrahydro-14-methylindolo[2‘3’-3，4Ipyrido[2，1-blquinazolin5-[71-11-one)是从传统中草药吴茱萸中提取的具有抗癌活性的药物，但其抗卵巢癌细胞活性的机制尚不清楚。本研究旨在探讨内质网应激蛋白PERK在EVO诱导人卵巢癌细胞凋亡中的作用机制。方法：采用四甲基偶氮唑盐比色法、DNA片段化分析和Giemsa染色进行细胞死亡分析。线粒体膜电位测定采用DiOC6染色。Western blotting分析蛋白质水平。结果：不同浓度的EVO对人卵巢癌细胞A2780、A2780CP、ES-2和SKOV-3的生长均有抑制作用，表现为细胞凋亡率增加，DNA梯状条带增多，caspase3和PARP蛋白裂解增加。在SKOV-3和A2780CP细胞中加入caspase抑制剂VAD和DEVD，并与JINK抑制剂SP600125(SP)和JINKI共同孵育细胞，但不能抑制PD98059、SB203530等其他MAPK和AKT抑制剂的作用，可显著抑制EVO诱导的人卵巢癌细胞的凋亡。此外，在EVO处理的人卵巢癌细胞中检测到磷酸化elF2α(pelF2α)和磷酸化PERK(PPERK)蛋白的表达增加，并被JINK抑制剂SP600125和JNKI抑制。应用PERK抑制剂GSK2606414对EVO诱导的人卵巢癌细胞A2780和A2780CP的凋亡有明显的保护作用。加入JNK或PERK抑制剂也可抑制线粒体膜电位的Evo破坏。构效关系研究表明，EVO第14位的烷基是通过激活JINK和PERK而诱导人卵巢癌细胞凋亡的重要途径。结论：为EVO通过激活JINK和PERK来阻断基质金属蛋白酶而诱导卵巢癌细胞的凋亡提供了证据，并且第14位的烷基是EVO的重要替代作用。(C)2015年爱思唯尔股份有限公司。版权所有。
Background: Evodiamine (EVO; 8,13,13b,14-tetrahydro-14-methylindolo[2'3'-3,4Ipyrido[2,1-blquinazolin5-[71-11-one derived from the traditional herbal medicine Evodia rutaecarpa was reported to possess anticancer activity; however, the anticancer mechanism of EVO against the viability of human ovarian cancer cells is still unclear.Purpose: A number of studies showed that chemotherapeutic benefits may result from targeting the endoplasmic reticular (ER) stress signaling pathway. The objective of the study is to investigate the mechanism by which ER stress protein PERK plays in EVO-induced apoptosis of human ovarian cancer cells. Methods: Cell death analysis was performed by MTT assay, DNA fragmentation assay, and Giemsa staining. DiOC6 staining was used for mitochondrial membrane potential measurement. Protein levels were analyzed by Western blotting. Pharmacological studies using MAPK inhibitors and PERK inhibitor GSK2606414 were involved.Results: The viability of human ovarian cancer cells A2780, A2780CP, ES-2, and SKOV-3 was inhibited by EVO at various concentrations in accordance with increases in the percentage of apoptotic cells, DNA ladders, and cleavage of caspase 3 and poly(ADP ribose) polymerase (PARP) proteins. Decreased viability of cells was reversed by adding caspase inhibitors VAD and DEVD in SKOV-3 and A2780CP cells, and incubation of cells with JINK inhibitor SP600125 (SP) and JINKI, but not other MAPK and AKT inhibitors including PD98059, SB203530, significantly prevented the apoptosis elicited by EVO in human ovarian cancer cells. Furthermore, increased expression of phospho-elF2 alpha (pelF2(alpha) and phospho-PERK (pPERK) proteins was detected in EVO-treated human ovarian cancer cells, and that was inhibited by adding JINK inhibitors SP600125 and JNKI. Application of a PERK inhibitor GSK2606414 showed a significant protection of human ovarian cancer cells A2780 and A2780CP from EVO-induced apoptosis. EVO disruption of mitochondrial membrane potential (MMP) was also inhibited by adding JNK or PERK inhibitors. The structure-activity relationship study indicated that the alkyl group at position 14 in EVO is important for apoptosis induction via activation of JINK and PERK in human ovarian cancer cells.Conclusion: Evidence supporting EVO induction of apoptosis via activation of JINK and PERK to disrupt MMP in human ovarian cancer cells is provided, and the alkyl at position 14 is a critical substitution for the apoptotic actions of EVO. (C) 2015 Elsevier GmbH. All rights reserved.