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首页> 外文期刊>Cell motility and the cytoskeleton >Molecular characterization of the effects of Y-27632.
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Molecular characterization of the effects of Y-27632.

机译:Y-27632作用的分子表征。

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Many key cellular functions, such as cell motility and cellular differentiation are mediated by Rho-associated protein kinases (ROCKs). Numerous studies have been conducted to examine the ROCK signal transduction pathways involved in these motile and contractile events with the aid of pharmacological inhibitors such as Y-27632. However the molecular mechanism of action of Y-27632 has not been fully defined. To assess the relative contribution of these Rho effectors to the effects of Y-27632, we compared the cytoskeletal phenotype, wound healing and neurite outgrowth in cells treated with Y-27632 or subjected to knockdown with ROCK-I, ROCK-II or PRK-2- specific siRNAs. Reduction of ROCK-I enhances the formation of thin actin-rich membrane extensions, a phenotype that closely resembles the effect of Y-27632. Knockdown of ROCK II or PRK-2, leads to the formation of disc-like extensions and thick actin bundles, respectively. The effect of ROCK-I knockdown also mimicked the effect of Y-27632 on wound closerrates. ROCK-I knockdown and Y-27632 enhanced wound closure rates, while ROCK-II and PRK-2 were not appreciably different from control cells. In neurite outgrowth assays, knockdown of ROCK-I, ROCK-II or PRK-2 enhances neurite lengths, however no individual knockdown stimulated neurite outgrowth as robustly as Y-27632. We conclude that several kinases contribute to the global effect of Y-27632 on cellular responses.
机译:Rho相关蛋白激酶(ROCKs)介导许多关键的细胞功能,例如细胞运动性和细胞分化。已经进行了许多研究,以借助诸如Y-27632的药理学抑制剂来检查参与这些运动和收缩事件的ROCK信号转导途径。但是,Y-27632的分子作用机理尚未完全阐明。为了评估这些Rho效应子对Y-27632的相对贡献,我们比较了用Y-27632处理或用ROCK-I,ROCK-II或PRK-敲除的细胞的细胞骨架表型,伤口愈合和神经突生长2-特异性siRNA。 ROCK-1的减少增强了富含肌动蛋白的薄膜延伸的形成,该表型非常类似于Y-27632的作用。敲低ROCK II或PRK-2分别导致形成盘状延伸物和厚肌动蛋白束。 ROCK-1敲低的作用还模仿了Y-27632对伤口闭合率的作用。 ROCK-I敲低和Y-27632增强了伤口闭合率,而ROCK-II和PRK-2与对照细胞没有明显差异。在神经突生长试验中,敲除ROCK-I,ROCK-II或PRK-2可以增强神经突的长度,但是没有单独的敲除刺激能够像Y-27632一样强烈地刺激神经突生长。我们得出结论,几种激酶有助于Y-27632对细胞反应的整体作用。

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