RRC ID 68594
著者 Kimura K, Teranishi S, Yamauchi J, Nishida T.
タイトル Role of JNK-dependent serine phosphorylation of paxillin in migration of corneal epithelial cells during wound closure.
ジャーナル Invest Ophthalmol Vis Sci
Abstract PURPOSE:Migration of corneal epithelial cells plays an important role in healing of corneal epithelial wounds. The role of c-Jun NH(2)-terminal kinase (JNK), a member of the family of mitogen-activated protein kinases, in the intracellular signaling responsible for the migration of corneal epithelial cells during wound closure was examined.
METHODS:Scratch wounds were introduced into cultured monolayers of simian virus 40-transformed human corneal epithelial (HCE) cells in the absence or presence of the JNK inhibitor SP600125. The phosphorylation and localization of JNK and paxillin during wound closure were examined by immunoblot and immunofluorescence analyses. The effects of a small interfering RNA (siRNA) specific for JNK and of a mutant form of paxillin on HCE cell migration were determined by transfection.
RESULTS:SP600125 inhibited wound healing in a time- and concentration-dependent manner. Immunoblot analysis showed that wounding increased the phosphorylation of JNK and of paxillin on serine (Ser) 178 in a manner sensitive to SP600125. Immunofluorescence staining revealed that phosphorylated JNK colocalized with paxillin at focal adhesions formed by HCE cells at the wound margin and that SP600125 inhibited the formation of such adhesions. Expression of JNK siRNA or of a paxillin mutant in which Ser178 is replaced by alanine inhibited HCE cell migration during wound closure.
CONCLUSIONS:JNK regulates HCE cell migration by modulating the phosphorylation of paxillin and the consequent formation of focal adhesions. A JNK-paxillin signaling pathway may thus play an important role in corneal epithelial wound healing in vivo.
巻・号 49(1)
ページ 125-32
公開日 2008-1-1
DOI 10.1167/iovs.07-0725
PII 49/1/125
PMID 18172084
MeSH Anthracenes / pharmacology Cell Movement / physiology* Cells, Cultured Dose-Response Relationship, Drug Enzyme Inhibitors / pharmacology Epithelium, Corneal / physiology* Fluorescent Antibody Technique, Indirect Humans Immunoblotting JNK Mitogen-Activated Protein Kinases / antagonists & inhibitors JNK Mitogen-Activated Protein Kinases / physiology* Microscopy, Fluorescence Paxillin / metabolism* Phosphorylation Plasmids RNA, Small Interfering / pharmacology Serine / metabolism* Signal Transduction Time Factors Transfection Wound Healing / drug effects Wound Healing / physiology*
IF 3.47
リソース情報
ヒト・動物細胞 HCE-T(RCB2280)