Raf-1/CK2 and RhoA/ROCK signaling promote TNF-α-mediated endothelial 0 apoptosis via regulating vimentin cytoskeleton
Raf-1/CK2 and RhoA/ROCK signaling promote TNF-α-mediated endothelial 0 apoptosis via regulating vimentin cytoskeleton
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DOI：
10.1016/j.tox.2017.07.010
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发表时间：
2017-08-15
期刊：
影响因子：
4.5
通讯作者：
Yao, Wenjuan
中科院分区：
文献类型：
--
作者：
Yang, Lifeng;Tang, Lian;Yao, Wenjuan
RhoA/ROCK和Raf-1/CK 2通路在细胞增殖、凋亡、分化和其他多种常见的细胞功能中发挥重要作用。我们以前报道，波形蛋白是负责TNF-α诱导的细胞凋亡。在此，我们研究了RhoA/ROCK和Raf-1/CK 2信号对波形蛋白丝和TNF-α介导的内皮细胞凋亡的调节。用TNF-α处理显著诱导RhoA和ROCK的活化以及ROCK 1的表达。RhoA缺陷可明显抑制TNF-α诱导的ROCK激活和ROCK 1表达。RhoA缺陷和ROCK活性抑制（Y-27632）都极大地抑制了TNF-α诱导的人脐静脉内皮细胞（HUVEC）中的内皮细胞凋亡并保持了细胞活力。RhoA抑制和ROCK抑制也明显减弱了TNF-α诱导的波形蛋白磷酸化和波形蛋白或磷酸波形蛋白的重塑。RhoA抑制和ROCK抑制通过降低caspase 3和8的活化显著抑制TNF-α介导的波形蛋白裂解。此外，TNF-α处理大大增强了Raf-1的活化。Raf-1或CK 2抑制剂（GW 5074或TBB）抑制Raf-1或CK 2可阻断波形蛋白磷酸化、重塑和内皮细胞凋亡，并在TNF-α诱导的HUVECs中保持细胞活力。然而，Raf-1抑制对TNF-α诱导的ROCK表达和活化没有显着影响，表明Raf-1/CK 2信号对波形蛋白的调节不依赖于ROCK。综上所述，这些结果表明RhoA/ROCK和Raf-1/CK 2途径均通过调节波形蛋白细胞骨架来负责TNF-α介导的内皮细胞毒性。
Both RhoA/ROCK and Raf-1/CK2 pathway play essential roles in cell proliferation, apoptosis, differentiation, and multiple other common cellular functions. We previously reported that vimentin is responsible for TNF-alpha-induced cell apoptosis. Herein, we investigated the regulation of RhoA/ROCK and Raf-1/CK2 signaling on vimentin filaments and endothelial apoptosis mediated by TNF-alpha. Treatment with TNF-alpha significantly induced the activation of RhoA and ROCK, and the expression of ROCK1. RhoA deficiency could obviously inhibit ROCK activation and ROCK1 expression induced by TNF-alpha. Both RhoA deficiency and ROCK activity inhibition (Y-27632) greatly inhibited endothelial apoptosis and preserved cell viability in TNF-alpha-induced human umbilical vein endothelial cells (HUVECs). Also vimentin phosphorylation and the remodeling of vimentin or phosphovimentin induced by TNF-alpha were obviously attenuated by RhoA suppression and ROCK inhibition. TNF-alpha-mediated vimentin cleavage was significantly inhibited by RhoA suppression and ROCK inhibition through decreasing the activation of caspase3 and 8. Furthermore, TNF-alpha treatment greatly enhanced the activation of Raf-1. Suppression of Raf-1 or CK2 by its inhibitor (GW5074 or TBB) blocked vimentin phosphorylation, remodeling and endothelial apoptosis, and preserved cell viability in TNF-alpha-induced HUVECs. However, Raf-1 inhibition showed no significant effect on TNF-alpha-induced ROCK expression and activation, suggesting that the regulation of Raf-1/CK2 signaling on vimentin was independent of ROCK. Taken together, these results indicate that both RhoA/ROCK and Raf-1/CK2 pathway are responsible for TNF-alpha-mediated endothelial cytotoxicity via regulating vimentin cytoskeleton.