Arp2/3抑制通过减少细胞骨架与膜的偶联和粘着斑组装,在MCF10A上皮细胞中诱导类阿米巴样突起。
Arp2/3 inhibition induces amoeboid-like protrusions in MCF10A epithelial cells by reduced cytoskeletal-membrane coupling and focal adhesion assembly.
作者信息
Beckham Yvonne, Vasquez Robert J, Stricker Jonathan, Sayegh Kareem, Campillo Clement, Gardel Margaret L
机构信息
Institute for Biophysical Dynamics, University of Chicago Medical Center, Chicago, Illinois, United States of America.
Section of Hematology, Oncology and Stem Cell Transplantation, Department of Pediatrics, University of Chicago Medical Center, Chicago, Illinois, United States of America.
出版信息
PLoS One. 2014 Jun 26;9(6):e100943. doi: 10.1371/journal.pone.0100943. eCollection 2014.
Here we demonstrate that Arp2/3 regulates a transition between mesenchymal and amoeboid protrusions in MCF10A epithelial cells. Using genetic and pharmacological means, we first show Arp2/3 inhibition impairs directed cell migration. Arp2/3 inhibition results in a dramatically impaired cell adhesion, causing deficient cell attachment and spreading to ECM as well as an 8-fold decrease in nascent adhesion assembly at the leading edge. While Arp2/3 does not play a significant role in myosin-dependent adhesion growth, mature focal adhesions undergo large scale movements against the ECM suggesting reduced coupling to the ECM. Cell edge protrusions occur at similar rates when Arp2/3 is inhibited but their morphology is dramatically altered. Persistent lamellipodia are abrogated and we observe a markedly increased incidence of blebbing and unstable pseuodopods. Micropipette-aspiration assays indicate that Arp2/3-inhibited cells have a weak coupling between the cell cortex and the plasma membrane, and suggest a potential mechanism for increased pseudopod and bleb formation. Pseudopods are not sensitive to reduced in formin or myosin II activity. Collectively, these results indicate that Arp2/3 is not necessary for rapid protrusion of the cell edge but plays a crucial role in assembling focal adhesions required for its stabilization.
在这里,我们证明了Arp2/3调节MCF10A上皮细胞中间质突起和阿米巴样突起之间的转变。通过基因和药理学方法,我们首先表明抑制Arp2/3会损害细胞的定向迁移。抑制Arp2/3会导致细胞黏附显著受损,导致细胞与细胞外基质(ECM)的附着和铺展不足,以及前沿新生黏附组装减少8倍。虽然Arp2/3在肌球蛋白依赖性黏附生长中不起重要作用,但成熟的黏着斑会相对于ECM进行大规模移动,这表明与ECM的耦合减少。当Arp2/3被抑制时,细胞边缘突起以相似的速率发生,但它们的形态发生了显著改变。持续性板状伪足消失,我们观察到气泡形成和不稳定伪足的发生率显著增加。微吸管抽吸试验表明,受Arp2/3抑制的细胞在细胞皮层和质膜之间的耦合较弱,并提示了伪足和气泡形成增加的潜在机制。伪足对formin或肌球蛋白II活性降低不敏感。总的来说,这些结果表明,Arp2/3对于细胞边缘的快速突起不是必需的,但在组装稳定所需的黏着斑中起关键作用。