MECHANISM OF ZIRAM-INDUCED APOPTOSIS IN HUMAN NATURAL KILLER CELLS
MECHANISM OF ZIRAM-INDUCED APOPTOSIS IN HUMAN NATURAL KILLER CELLS
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DOI：
10.1177/039463201202500406
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发表时间：
2012-10-01
影响因子：
3.5
通讯作者：
Kawada, T.
中科院分区：
文献类型：
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作者：
Li, Q.;Kobayashi, M.;Kawada, T.
我们之前发现齐拉姆（ziram）是一种二硫代氨基甲酸盐类杀菌剂，能够以剂量依赖性方式显着抑制自然杀伤（NK）活性。为了探索这种抑制的机制，我们研究了齐拉姆诱导的人类 NK 细胞凋亡。用0.0625-4μM的ziram处理人NK-92MI细胞2-64小时。通过 FITC-Annexin-V/PI 染色测定细胞凋亡。为了探讨细胞凋亡的机制，通过流式细胞术测定细胞内活性半胱天冬酶 3、3/7、8 和 9 的水平以及泛半胱天冬酶和线粒体细胞色素-c 的释放。使用 MitoLight (TM) 细胞凋亡检测试剂盒测定线粒体跨膜电位的破坏。研究发现，ziram 以剂量和时间依赖性方式诱导人类 NK 细胞凋亡。 Ziram 以剂量依赖性方式增加细胞内活性 caspase 3、3/7、8 和 9 以及泛 caspase 的水平，并且 caspase-3 抑制剂 Z-DEVD-FMK 和通用 caspase 抑制剂 Z-VAD-FMK 部分但显着抑制细胞凋亡。 Ziram 还破坏线粒体跨膜电位，并以剂量依赖性方式引起线粒体细胞色素-c 释放。这些发现表明，ziram可以诱导人NK细胞凋亡，并且凋亡至少是由caspase-cascade和线粒体/细胞色素-c途径介导的。
We previously found that ziram, a dithiocarbamate fungicide, significantly inhibited natural killer (NK) activity in a dose-dependent manner. To explore the mechanism of this inhibition, we investigated ziram-induced apoptosis in human NK cells. Human NK-92MI cells were treated with ziram at 0.0625-4 mu M for 2-64 h. Apoptosis was determined by FITC-Annexin-V/PI staining. To explore the mechanism of apoptosis, intracellular levels of active caspases 3, 3/7, 8, and 9 and pan-caspase and mitochondrial cytochrome-c release were determined by flow cytometry. Disruption to mitochondrial transmembrane potential was determined with a MitoLight (TM) Apoptosis Detection Kit. It was found that ziram induced apoptosis in a dose- and time-dependent manner in human NK cells. Ziram increased the intracellular levels of active caspases 3, 3/7, 8, and 9 and pan-caspase in a dose-dependent manner, and a caspase-3 inhibitor, Z-DEVD-FMK, and a general caspase inhibitor, Z-VAD-FMK, partially but significantly inhibited the apoptosis. Ziram also disrupted mitochondrial transmembrane potential and caused mitochondrial cytochrome-c release in a dose-dependent manner. These findings indicate that ziram can induce apoptosis in human NK cells, and the apoptosis is at least mediated by both the caspase-cascade and the mitochondria/cytochrome-c pathways.