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Dissecting the roles of ROCK isoforms in stress-induced cell detachment

机译:剖析ROCK亚型在应激诱导的细胞脱离中的作用

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The homologous Rho kinases, ROCK1 and ROCK2, are involved in stress fiber assembly and cell adhesion and are assumed to be functionally redundant. Using mouse embryonic fibroblasts (MEFs) derived from ROCK1-/- and ROCK2-/- mice, we have recently reported that they play different roles in regulating doxorubicin-induced stress fiber disassembly and cell detachment: ROCK1 is involved in destabilizing the actin cytoskeleton and cell detachment, whereas ROCK2 is required for stabilizing the actin cytoskeleton and cell adhesion. Here, we present additional insights into the roles of ROCK1 and ROCK2 in regulating stress-induced impairment of cell-matrix and cell-cell adhesion. In response to doxorubicin, ROCK1-/- MEFs showed significant preservation of both focal adhesions and adherens junctions, while ROCK2-/- MEFs exhibited impaired focal adhesions but preserved adherens junctions compared with the wild-type MEFs. Additionally, inhibition of focal adhesion or adherens junction formations by chemical inhibitors abolished the anti-detachment effects of ROCK1 deletion. Finally, ROCK1-/- MEFs, but not ROCK2-/- MEFs, also exhibited preserved central stress fibers and reduced cell detachment in response to serum starvation. These results add new insights into a novel mechanism underlying the anti-detachment effects of ROCK1 deletion mediated by reduced peripheral actomyosin contraction and increased actin stabilization to promote cell-cell and cell-matrix adhesion. Our studies further support the differential roles of ROCK isoforms in regulating stress-induced loss of central stress fibers and focal adhesions as well as cell detachment.
机译:同源Rho激酶ROCK1和ROCK2参与应力纤维的组装和细胞粘附,并被认为在功能上是多余的。最近,我们报道了使用源自ROCK1-/-和ROCK2-/-小鼠的小鼠胚胎成纤维细胞(MEF),它们在调节阿霉素诱导的应力纤维分解和细胞脱离中起着不同的作用:ROCK1参与破坏肌动蛋白细胞骨架和细胞分离,而ROCK2是稳定肌动蛋白细胞骨架和细胞粘附所必需的。在这里,我们提出了ROCK1和ROCK2在调节应激诱导的细胞基质损伤和细胞粘附方面的作用的其他见解。响应阿霉素,ROCK1-/-MEFs显着保留了粘着斑和黏附连接,而ROCK2-/-MEFs表现出了局灶性粘连受损,但与野生型MEF相比,保留了粘连。此外,通过化学抑制剂抑制粘着斑或粘附连接的形成消除了ROCK1缺失的抗分离作用。最后,ROCK1-/-MEFs,但不是ROCK2-/-MEFs,也表现出对血清饥饿的响应而保留的中心应力纤维和减少的细胞脱离。这些结果增加了对新型机制的新见解,该机制是由减少的外周放线菌素收缩和增加的肌动蛋白稳定作用介导的ROCK1缺失的抗分离作用,从而促进细胞与细胞和基质的粘附。我们的研究进一步支持了ROCK亚型在调节应力诱导的中央应力纤维和粘着斑的粘连以及细胞脱离中的不同作用。

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