RRC ID 44084
Author Felkl M, Tomas K, Smid M, Mattes J, Windoffer R, Leube RE.
Title Monitoring the cytoskeletal EGF response in live gastric carcinoma cells.
Journal PLoS One
Abstract Altered cell motility is considered to be a key factor in determining tumor invasion and metastasis. Epidermal growth factor (EGF) signaling has been implicated in this process by affecting cytoskeletal organization and dynamics in multiple ways. To sort the temporal and spatial regulation of EGF-dependent cytoskeletal re-organization in relation to a cell's motile behavior time-lapse microscopy was performed on EGF-responsive gastric carcinoma-derived MKN1 cells co-expressing different fluorescently labeled cytoskeletal filaments and focal adhesion components in various combinations. The experiments showed that EGF almost instantaneously induces a considerable increase in membrane ruffling and lamellipodial activity that can be inhibited by Cetuximab EGF receptor antibodies and is not elicited in non-responsive gastric carcinoma Hs746T cells. The transient cell extensions are rich in actin but lack microtubules and keratin intermediate filaments. We show that this EGF-induced increase in membrane motility can be measured by a simple image processing routine. Microtubule plus-ends subsequently invade growing cell extensions, which start to accumulate focal complexes at the lamellipodium-lamellum junction. Such paxillin-positive complexes mature into focal adhesions by tyrosine phosphorylation and recruitment of zyxin. These adhesions then serve as nucleation sites for keratin filaments which are used to enlarge the neighboring peripheral keratin network. Focal adhesions are either disassembled or give rise to stable zyxin-rich fibrillar adhesions which disassemble in the presence of EGF to support formation of new focal adhesion sites in the cell periphery. Taken together the results serve as a basis for modeling the early cytoskeletal EGF response as a tightly coordinated and step-wise process which is relevant for the prediction of the effectiveness of anti-EGF receptor-based tumor therapy.
Volume 7(9)
Pages e45280
Published 2012-1-1
DOI 10.1371/journal.pone.0045280
PII PONE-D-12-15066
PMID 23028903
PMC PMC3459943
MeSH Actins / genetics Actins / metabolism Antibodies, Monoclonal / pharmacology* Antibodies, Monoclonal, Humanized Antineoplastic Agents / pharmacology* Carcinoma / drug therapy Carcinoma / genetics Carcinoma / pathology Cell Movement / drug effects Cetuximab Epidermal Growth Factor / pharmacology ErbB Receptors / antagonists & inhibitors* ErbB Receptors / genetics Focal Adhesions / drug effects* Focal Adhesions / genetics Focal Adhesions / ultrastructure Gene Expression / drug effects Humans Keratins / genetics Keratins / metabolism Microtubules / drug effects* Microtubules / genetics Microtubules / ultrastructure Paxillin / genetics Paxillin / metabolism Phosphorylation Pseudopodia / drug effects* Pseudopodia / genetics Pseudopodia / ultrastructure Signal Transduction / drug effects Stomach Neoplasms / drug therapy Stomach Neoplasms / genetics Stomach Neoplasms / pathology Time Factors Time-Lapse Imaging Tumor Cells, Cultured Zyxin / genetics Zyxin / metabolism
IF 2.74
Times Cited 12
Human and Animal Cells MKN1(RCB1003)