当前位置： SCI文献检索 > JAPANESE JOURNAL OF OPHTHALMOLOGY期刊下所有文献 > Differential regulation of focal adhesion kinase and paxillin phosphorylation by the small GTP-binding protein Rho in human corneal epithelial cells.

Differential regulation of focal adhesion kinase and paxillin phosphorylation by the small GTP-binding protein Rho in human corneal epithelial cells.

Abstract：

PURPOSE:Lysophosphatidic acid (LPA), which activates the small guanosine triphosphate (GTP)-binding protein Rho, was previously shown to promote migration of the rabbit corneal epithelium in culture. The signaling pathway responsible for this effect of LPA was examined in this study with a human corneal epithelial (HCE) cell line. METHODS:The activation of Rho was detected with a pull-down assay. Tyrosine phosphorylation of paxillin and of focal adhesion kinase (FAK) were examined both by immunofluorescence staining and by immunoprecipitation and immunoblot analyses. Expression of integrins alpha5 and beta1 was evaluated by immunoblot analysis. RESULTS:Incubation of cells with LPA (10 micro M) for 2 min resulted in marked activation of Rho, and this effect was blocked by pretreatment with the Rho inhibitor exoenzyme C3 (2 micro g/ml) for 24 h. Tyrosine phosphorylation of both paxillin and FAK was detected in HCE cells under basal conditions by immunofluorescence staining, immunoprecipitation, and immunoblot analyses. LPA induced a concentration- and time-dependent increase in the tyrosine phosphorylation of paxillin, which was maximal at a concentration of 10 micro M and a time of 2 min. Exoenzyme C3 inhibited LPA-induced paxillin phosphorylation. Neither LPA nor exoenzyme C3 affected tyrosine phosphorylation of FAK or expression of integrins alpha5 and beta1. CONCLUSIONS:LPA induces Rho activation and the consequent tyrosine phosphorylation of paxillin in HCE cells, and these effects likely contribute to the promotion of corneal epithelial migration by this agent.

journal_name

Jpn J Ophthalmol

journal_title

Japanese journal of ophthalmology

authors

Saito J,Morishige N,Chikama T,Gu J,Sekiguchi K,Nishida T

doi

10.1007/s10384-003-0059-2

keywords:

subject

Has Abstract

pub_date

2004-05-01 00:00:00

pages

199-207

issue

3

eissn

0021-5155

issn

1613-2246

journal_volume

48

pub_type

杂志文章

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