Maria Hägg Olofsson, Aleksandra Mandic Havelka, Slavica Brnjic, Maria C Shoshan, Stig Linder
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引用次数: 56
摘要
背景:细胞内游离钙([Ca2+]i)是凋亡信号传导的关键因素,许多钙依赖的凋亡途径已经被描述。我们在这里使用化学生物学策略来阐明这些不同途径的相对重要性。结果:通过筛选化学文库,首次鉴定出40种诱导细胞凋亡的药物(“生物探针”)。使用p53, AP-1, NFAT和NF-kappaB报告细胞系,这些生物探针被证实可以诱导不同的信号传导模式。使用钙螯合剂BAPTA-AM的实验表明,Ca2+参与了大多数生物探针诱导细胞凋亡的过程,并且Ca2+通常需要进入细胞凋亡过程数小时。进一步的研究表明,钙调蛋白通路是细胞凋亡反应的重要介质。与抑制calpain、calcineurin/PP2B或DAP激酶相比,抑制calmodulin kinase II (CaMKII)能更有效地抑制细胞凋亡。我们使用其中一种生物探针,植物生物碱helenalin,来研究CaMKII在细胞凋亡中的作用。Helenalin诱导CaMKII、ASK1和jun - n-末端激酶(JNK)活性,抑制这些激酶可抑制细胞凋亡。结论:我们的研究表明,钙信号通常不是凋亡过程中的早期事件,并提示CaMKII/ASK1信号机制对于某些类型刺激下JNK的持续激活和凋亡是重要的。
Charting calcium-regulated apoptosis pathways using chemical biology: role of calmodulin kinase II.
Background: Intracellular free calcium ([Ca2+]i) is a key element in apoptotic signaling and a number of calcium-dependent apoptosis pathways have been described. We here used a chemical biology strategy to elucidate the relative importance of such different pathways.
Results: A set of 40 agents ("bioprobes") that induce apoptosis was first identified by screening of a chemical library. Using p53, AP-1, NFAT and NF-kappaB reporter cell lines, these bioprobes were verified to induce different patterns of signaling. Experiments using the calcium chelator BAPTA-AM showed that Ca2+ was involved in induction of apoptosis by the majority of the bioprobes and that Ca2+ was in general required several hours into the apoptosis process. Further studies showed that the calmodulin pathway was an important mediator of the apoptotic response. Inhibition of calmodulin kinase II (CaMKII) resulted in more effective inhibition of apoptosis compared to inhibition of calpain, calcineurin/PP2B or DAP kinase. We used one of the bioprobes, the plant alkaloid helenalin, to study the role of CaMKII in apoptosis. Helenalin induced CaMKII, ASK1 and Jun-N-terminal kinase (JNK) activity, and inhibition of these kinases inhibited apoptosis.
Conclusion: Our study shows that calcium signaling is generally not an early event during the apoptosis process and suggests that a CaMKII/ASK1 signaling mechanism is important for sustained JNK activation and apoptosis by some types of stimuli.