Cell–cell contacts protect against t-BuOOH-induced cellular damage and ferroptosis in vitro
Cell–cell contacts protect against t-BuOOH-induced cellular damage and ferroptosis in vitro
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DOI：
10.1007/s00204-019-02413-w
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发表时间：
2019-02
影响因子：
6.1
通讯作者：
Christine Wenz;D. Faust;Berenike Linz;Christian Turmann;T. Nikolova;C. Dietrich
中科院分区：
文献类型：
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作者：
Christine Wenz;D. Faust;Berenike Linz;Christian Turmann;T. Nikolova;C. Dietrich
铁凋亡是最近发现的依赖于铁和脂质过氧化的调节性坏死途径。由于它是克服肿瘤化疗中细胞凋亡抵抗的一种很有前途的方法，因此受到了广泛的关注。我们最近发现叔丁基过氧化氢（t-BuOOH）是一种新的铁凋亡诱导剂。t-BuOOH是一种广泛使用的体外诱导氧化应激的化合物。t-BuOOH诱导小鼠和人细胞系的脂质过氧化，从而导致铁凋亡。t-BuOOH还导致线粒体膜电位的损失，DNA双链断裂的形成和复制阻滞。在这里，我们专门解决的问题是否细胞接触调节BuOOH诱导的铁凋亡和细胞损伤。为此，将鼠NIH 3 T3或人HaCaT细胞接种至汇合，但低于其饱和密度，以允许建立细胞与细胞接触而不诱导静止。然后用t-BuOOH（分别为50或200 µM）处理细胞。我们发现，细胞与细胞的接触可以减少基础和叔丁氧醇引发的脂质过氧化反应，从而阻止铁细胞凋亡。使用特异性铁凋亡诱导剂erastin也获得了类似的结果。细胞-细胞接触进一步防止BuOOH诱导的线粒体膜电位损失和DNA双链断裂的形成。有趣的是，细胞-细胞接触未能阻止BuOOH介导的复制阻滞或氧化性基底损伤8-氧代-dG的形成。由于（i）用过氧化氢、甲磺酸甲酯或UV-C处理后观察到细胞死亡保护的证据，以及（ii）在几种细胞系中观察到的证据，我们得出结论，细胞-细胞接触的保护是一种普遍现象。细胞接触对毒性的影响可能在癌症化疗中具有重要意义。
Ferroptosis is a recently discovered pathway of regulated necrosis dependent on iron and lipid peroxidation. It has gained broad attention since it is a promising approach to overcome resistance to apoptosis in cancer chemotherapy. We have recently identifiedtertiary-butyl hydroperoxide (t-BuOOH) as a novel inducer of ferroptosis.t-BuOOH is a widely used compound to induce oxidative stress in vitro.t-BuOOH induces lipid peroxidation and consequently ferroptosis in murine and human cell lines.t-BuOOH additionally results in a loss of mitochondrial membrane potential, formation of DNA double-strand breaks, and replication block. Here, we specifically address the question whether cell–cell contacts regulatet-BuOOH-induced ferroptosis and cellular damage. To this end, murine NIH3T3 or human HaCaT cells were seeded to confluence, but below their saturation density to allow the establishment of cell–cell contacts without inducing quiescence. Cells were then treated witht-BuOOH (50 or 200 µM, respectively). We revealed that cell–cell contacts reduce basal andt-BuOOH-triggered lipid peroxidation and consequently block ferroptosis. Similar results were obtained with the specific ferroptosis inducer erastin. Cell–cell contacts further protect againstt-BuOOH-induced loss of mitochondrial membrane potential, and formation of DNA double-strand breaks. Interestingly, cell–cell contacts failed to preventt-BuOOH-mediated replication block or formation of the oxidative base lesion 8-oxo-dG. Since evidence of protection against cell death was both (i) observed after treatment with hydrogen peroxide, methyl methanesulfonate or UV-C, and (ii) seen in several cell lines, we conclude that protection by cell–cell contacts is a widespread phenomenon. The impact of cell–cell contacts on toxicity might have important implications in cancer chemotherapy.