RAS-RAF-MEK-dependent oxidative cell death involving voltage-dependent anion channels
RAS-RAF-MEK-dependent oxidative cell death involving voltage-dependent anion channels
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DOI：
10.1038/nature05859
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发表时间：
2007-06-14
期刊：
影响因子：
64.8
通讯作者：
Stockwell, Brent R.
中科院分区：
文献类型：
--
作者：
Yagoda, Nicholas;von Rechenberg, Moritz;Stockwell, Brent R.
区分正常细胞和肿瘤细胞的基因组成的治疗方法对于治疗和理解癌症是有价值的。具有癌基因选择性致死性的小分子可能揭示癌蛋白的新功能，并能够创造更具选择性的药物（1）。在这里，我们描述了选择性抗肿瘤剂erastin的作用机制，涉及RAS-RAF- MEK信号通路在细胞增殖，分化和存活中的作用。Erastin在癌基因HRAS、KRAS或BRAF中携带突变的人肿瘤细胞中表现出更大的致死性。利用亲和纯化和质谱分析，我们发现erastin通过线粒体电压依赖性阴离子通道（VDACs）发挥作用-这是抗癌药物的一个新靶点。我们表明，erastin治疗的细胞窝藏致癌RAS导致氧化物种的外观和随后的死亡，通过氧化，非凋亡机制。RNA干扰介导的VDAC 2或VDAC 3的敲低导致对erastin的抗性，这两种VDAC亚型与erastin的作用机制有关。此外，使用纯化的线粒体表达一个单一的VDAC亚型，我们发现erastin改变外线粒体膜的通透性。最后，使用放射性标记的类似物和过滤器结合试验，我们表明erastin直接结合VDAC 2。这些结果表明，VDAC蛋白的配体可以在RAS-RAF- MEK途径中携带激活突变的一些肿瘤细胞中选择性地诱导非凋亡性细胞死亡。
Therapeutics that discriminate between the genetic makeup of normal cells and tumour cells are valuable for treating and understanding cancer. Small molecules with oncogene-selective lethality may reveal novel functions of oncoproteins and enable the creation of more selective drugs(1). Here we describe the mechanism of action of the selective anti-tumour agent erastin, involving the RAS - RAF - MEK signalling pathway functioning in cell proliferation, differentiation and survival. Erastin exhibits greater lethality in human tumour cells harbouring mutations in the oncogenes HRAS, KRAS or BRAF. Using affinity purification and mass spectrometry, we discovered that erastin acts through mitochondrial voltage-dependent anion channels (VDACs) - a novel target for anti-cancer drugs. We show that erastin treatment of cells harbouring oncogenic RAS causes the appearance of oxidative species and subsequent death through an oxidative, non-apoptotic mechanism. RNA-interference-mediated knockdown of VDAC2 or VDAC3 caused resistance to erastin, implicating these two VDAC isoforms in the mechanism of action of erastin. Moreover, using purified mitochondria expressing a single VDAC isoform, we found that erastin alters the permeability of the outer mitochondrial membrane. Finally, using a radiolabelled analogue and a filter-binding assay, we show that erastin binds directly to VDAC2. These results demonstrate that ligands to VDAC proteins can induce non-apoptotic cell death selectively in some tumour cells harbouring activating mutations in the RAS - RAF - MEK pathway.