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PLoS One. 2014 Jun 26;9(6):e100943. doi: 10.1371/journal.pone.0100943. eCollection 2014.

Arp2/3 inhibition induces amoeboid-like protrusions in MCF10A epithelial cells by reduced cytoskeletal-membrane coupling and focal adhesion assembly.

Author information

1
Institute for Biophysical Dynamics, University of Chicago Medical Center, Chicago, Illinois, United States of America.
2
Section of Hematology, Oncology and Stem Cell Transplantation, Department of Pediatrics, University of Chicago Medical Center, Chicago, Illinois, United States of America.
3
James Franck Institute, University of Chicago, Chicago, Illinois, United States of America; Department of Physics, University of Chicago, Chicago, Illinois, United States of America.
4
Laboratoire Physico-Chimie, Institut Curie, Centre de Recherche, Paris, France; Laboratoire Analyse et Modélisation pour la Biologie et l' Environnement, Université d'Evry Val d'Essonne, Evry, France.
5
Institute for Biophysical Dynamics, University of Chicago Medical Center, Chicago, Illinois, United States of America; James Franck Institute, University of Chicago, Chicago, Illinois, United States of America; Department of Physics, University of Chicago, Chicago, Illinois, United States of America.

Abstract

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.

PMID:
24967897
PMCID:
PMC4072770
DOI:
10.1371/journal.pone.0100943
[Indexed for MEDLINE]
Free PMC Article

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