Caspases-2, -3, and -7 are involved in thapsigargin-induced apoptosis of SH-SY5Y neuroblastoma cells
Caspases-2, -3, and -7 are involved in thapsigargin-induced apoptosis of SH-SY5Y neuroblastoma cells
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DOI：
10.1002/jnr.20471
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发表时间：
2005-05-15
影响因子：
4.2
通讯作者：
Dahmer, MK
中科院分区：
文献类型：
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作者：
Dahmer, MK
据报道，Caspase-2 在许多不同条件下观察到的细胞死亡中发挥作用；然而，尚不清楚 caspase-2 是否在内质网 (ER) 应激引发的细胞死亡中发挥作用。本研究的目的是确定 caspase-2 是否参与毒胡萝卜素诱导的 ER 应激引起的 SH-SY5Y 神经母细胞瘤细胞死亡。毒胡萝卜素处理（1 μM，16 小时）刺激了 caspase-2、-3 和 -7 的蛋白水解过程，表明这些 caspase 是由 ER 应激激活的。通过使用细胞可渗透的半胱天冬酶抑制剂来检查这些半胱天冬酶在毒胡萝卜素诱导的细胞死亡中的作用。在没有使用 caspase 抑制剂进行预处理的情况下，毒胡萝卜素（0.1 μM，20 小时）将活细胞数量减少至起始时间对照的 53.9% +/- 3.3%。用泛 caspase 抑制剂 Z-VAD-FIVIK 或 caspase-2 选择性抑制剂 Z-VDVAD-FMK 预处理 90 分钟可抑制毒胡萝卜素刺激的细胞死亡，导致活细胞数分别为 115.6% +/- 5.3% (P < 0.001) 和 69.3% +/- 2.9% (P < 0.01)。启动时间控制。 caspase-3-和-7-选择性抑制剂 Z-DEVD-FMK 和 caspase-9-选择性抑制剂 Z-LEHD-FMK 均未显着影响毒胡萝卜素刺激的细胞死亡。在 SH-SY5Y 细胞中还鉴定出抗天冬氨酸蛋白酶 12 反应蛋白，但毒胡萝卜素对该蛋白的蛋白水解没有影响。这些数据表明 caspase-2、-3 和 -7 参与内质网应激介导的 SH-SY5Y 细胞死亡。 (c) 2005 年 Wiley-Liss, Inc.
Caspase-2 has been reported to play a role in the cell death observed under a number of different conditions; however, it is unclear whether caspase-2 plays a role in cell death triggered by endoplasmic reticulum (ER) stress. The purpose of this study was to determine whether caspase-2 is involved in SH-SY5Y neuroblastoma cell death caused by thapsigargin-induced ER stress. Thapsigargin treatment (1 mu M, 16 hr) stimulated the proteolytic processing of caspases-2, -3, and -7, suggesting that these caspases are activated by ER stress. The role of these caspases in thapsigargin-induced cell death was examined by using cell-permeable caspase inhibitors. In the absence of pretreatment with caspase inhibitors, thapsigargin (0.1 mu M, 20 hr) reduced the number of viable cells to 53.9% +/- 3.3% of starting-time control. Pretreatment for 90 min with either the pan-caspase inhibitor Z-VAD-FIVIK or the caspase-2-selective inhibitor Z-VDVAD-FMK inhibited thapsigargin-stimulated cell death, resulting in the number of viable cells being 115.6% +/- 5.3% (P < 0.001) and 69.3% +/- 2.9% (P < 0.01), respectively, of starting-time control. Neither the caspase-3- and -7-selective inhibitor Z-DEVD-FMK nor the caspase-9-selective inhibitor Z-LEHD-FMK significantly affected thapsigargin-stimulated cell death. An anticaspase-12-reactive protein was also identified in SH-SY5Y cells, but thapsigargin had no effect on proteolysis of this protein. These data demonstrate that caspases-2, -3, and -7 are involved in ER stress-mediated death of SH-SY5Y cells. (c) 2005 Wiley-Liss, Inc.