Gelsolin, but not its cleavage, is required for TNF-induced ROS generation and apoptosis in MCF-7 cells.
Gelsolin, but not its cleavage, is required for TNF-induced ROS generation and apoptosis in MCF-7 cells.
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TNF 诱导的 MCF-7 细胞中 ROS 生成和细胞凋亡需要凝溶胶蛋白，而不是其裂解。
DOI：
10.1016/j.bbrc.2009.05.078
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发表时间：
2009-07
期刊：
影响因子：
--
通讯作者：
李勤喜
中科院分区：
文献类型：
--
作者：
李勤喜
肿瘤坏死因子（tumor necrosis factor，TNF）可诱导包括MCF-7细胞在内的多种细胞凋亡。为了确定负责TNF诱导的细胞凋亡的基因，我们通过逆转录病毒插入介导的随机突变产生了一系列TNF抗性MCF-7细胞系。在其中一个抗性系中，发现凝溶胶蛋白因病毒插入而被破坏。在该突变细胞系（Gelmut）中外源表达凝溶胶蛋白恢复了对TNF诱导的细胞死亡的敏感性，并且通过siRNA敲低凝溶胶蛋白赋予MCF-7细胞对TNF的抗性，表明凝溶胶蛋白是TNF诱导的细胞凋亡所需的。有趣的是，Gelmutcells对凋亡诱导的抗性对TNF具有选择性，因为Gelmutand野生型细胞对测试的其他死亡刺激物显示出相似的敏感性。此外，在Gelmutcells中TNF诱导的ROS产生显著降低，表明凝溶胶蛋白介导的ROS产生在TNF诱导的MCF-7细胞凋亡中起着至关重要的作用。重要的是，半胱天冬酶介导的凝溶胶蛋白裂解是TNF触发的ROS产生和随后的MCF-7细胞凋亡的关键。因此，我们的研究提供了遗传学证据连接凝溶胶蛋白介导的活性氧产生肿瘤坏死因子诱导的细胞死亡。
The tumor necrosis factor (TNF) can induce apoptosis in many cells including MCF-7 cells. To identify the genes responsible for TNF-induced apoptosis, we generated a series of TNF-resistant MCF-7 cell lines by employing retrovirus insertion-mediated random mutagenesis. In one of the resistant lines, gelsolin was found to be disrupted by viral insertion. Exogenous expression of gelsolin in this mutant cell line (Gelmut) restored the sensitivity to TNF-induced cell death and knock-down of gelsolin by siRNA conferred MCF-7 cells with resistance to TNF, indicating that gelsolin is required for TNF-induced apoptosis. Interestingly, the resistance of Gelmutcells to apoptosis induction is selective to TNF, since Gelmutand wild-type cells showed similar sensitivity to other death stimuli that were tested. Furthermore, TNF-induced ROS production in Gelmutcells was significantly decreased, demonstrating that gelsolin-mediated ROS generation plays a crucial role in TNF-induced apoptosis in MCF-7 cells. Importantly, caspase-mediated gelsolin cleavage is dispensable for TNF-triggered ROS production and subsequent apoptosis of MCF-7 cells. Our study thus provides genetic evidence linking gelsolin-mediated ROS production to TNF-induced cell death.
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DOI：
10.11501/3169202
发表时间：
2000-03
期刊：
--
影响因子：
--
作者：
草野 元
通讯作者：
草野 元
影响因子：
8
作者：
L. Müllauer;H. Fujita;A. Ishizaki;N. Kuzumaki
通讯作者：
L. Müllauer;H. Fujita;A. Ishizaki;N. Kuzumaki
影响因子：
5.2
作者：
S. Moriya;K. Yanagihara;H. Fujita;N. Kuzumaki
通讯作者：
S. Moriya;K. Yanagihara;H. Fujita;N. Kuzumaki
影响因子：
8
作者：
Kusano, H;Shimizu, S;Tsujimoto, Y
通讯作者：
Tsujimoto, Y
影响因子：
4
作者：
Li, JQ;Li, QX;Han, JH
通讯作者：
Han, JH