RRC ID |
44084
|
著者 |
Felkl M, Tomas K, Smid M, Mattes J, Windoffer R, Leube RE.
|
タイトル |
Monitoring the cytoskeletal EGF response in live gastric carcinoma cells.
|
ジャーナル |
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.
|
巻・号 |
7(9)
|
ページ |
e45280
|
公開日 |
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
|
引用数 |
12
|
WOS 分野
|
CELL BIOLOGY
|
リソース情報 |
ヒト・動物細胞 |
MKN1(RCB1003) |