Ikarugamycin induces DNA damage, intracellular calcium increase, p38 MAP kinase activation and apoptosis in HL-60 human promyelocytic leukemia cells
Ikarugamycin induces DNA damage, intracellular calcium increase, p38 MAP kinase activation and apoptosis in HL-60 human promyelocytic leukemia cells
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DOI：
10.1016/j.mrfmmm.2011.03.001
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发表时间：
2011-05-10
影响因子：
2.3
通讯作者：
Kopp, Brigitte
中科院分区：
文献类型：
--
作者：
Popescu, Ruxandra;Heiss, Elke Hannelore;Kopp, Brigitte
斑鸠霉素 (IKA) 是一种具有强抗原虫和细胞毒活性的抗生素。我们工作的目的是深入了解 IKA 在 HL-60 白血病细胞中细胞毒性作用的作用机制，以评估其作为抗肿瘤药物的潜力。 IKA 反应后细胞活力降低（IC50 为 221.3 nM），而 24 小时后，具有亚二倍体 DNA 含量的 HL-60 细胞数量显着增加。使用免疫印迹通过 caspase-9、-8 和 -3 的裂解证实细胞凋亡。单细胞凝胶电泳表明存在早期基因毒性效应。通过 Fluo-3-AM 荧光的流式细胞术分析监测细胞内钙 ([Ca2+](i)) 水平，表明胞质钙的增加与半胱天冬酶的裂解相关。此外，IKA 触发了 p38 MAP 激酶的激活，该激酶部分依赖于升高的 [Ca2+](i) 浓度并有助于 caspase 激活。数据表明，IKA 通过基因毒性和 caspase 激活诱导 HL-60 细胞凋亡，这部分与细胞内钙水平的增加和 p38 MAP 激酶的激活相关。 (C) 2011 Elsevier B.V. 保留所有权利。
Ikarugamycin (IKA) is an antibiotic with strong antiprotozoal and cytotoxic activity. The purpose of our work was to provide insight into the mechanism of action characterizing the cytotoxic effect of IKA in HL-60 leukemia cells in order to evaluate its potential as an antineoplastic agent. Cell viability was reduced in response to IKA (IC50 of 221.3 nM), while the amount of HL-60 cells with a subdiploid DNA content increased significantly after 24 h. Apoptotic cell death was confirmed by the cleavage of caspase-9, -8 and -3 using immunoblotting. Single cell gel electrophoresis pointed to an early genotoxic effect. Monitoring of intracellular calcium ([Ca2+](i)) levels by flow cytometric analysis of Fluo-3-AM fluorescence indicated an increase in cytosolic calcium that correlated with the cleavage of caspases. In addition, IKA triggered the activation of p38 MAP kinase which was partly dependent on elevated [Ca2+](i) concentrations and contributed to caspase activation. The data demonstrate that IKA induced apoptosis in HL-60 cells through genotoxicity and caspase activation which was in part correlated to an increase in intracellular calcium levels and activation of p38 MAP kinase. (C) 2011 Elsevier B.V. All rights reserved.