Mitomycin C retardation of corneal fibroblast migration via sustained dephosphorylation of paxillin at tyrosine 118.

摘要: PURPOSE To investigate how mitomycin C (MMC) modulates corneal fibroblast migration and its molecular mechanisms in the wound healing process. METHODS After treatment with 0 0.2 mg · mL(-1) MMC for 5 minutes, effect of on cell human fibroblasts (HCFs) was examined a assay. Both focal adhesion kinase (FAK) paxillin (PXN) expressions HCFs were analyzed by semiquantitative real-time PCR, immunoblotting, immunofluorescence confocal microscopy. Using gene silencing or overexpression lentiviral-based pseudovirion infection, phosphorylation level FAK, PXN, mutated PXNs at tyrosine sites 31 (Y31F-EGFP) 118 (Y118F-EGFP) verified HCFs. RESULTS retarded HCF 1 2 days posttreatment (dpt). reduced levels FAK transcript protein, but increased both protein expression PXN dpt. Furthermore, upregulated FAK-pY397, which subsequently enhanced PXN-pY31 dose-dependent manner Concurrently, downregulated PXN-pY118 However, resulted dephosphorylation PXN-pY31, The FAK/PXN complex MMC-treated detected more than leading edge dpt, contributing to retardation migration. Y118F-EGFP-expressing exhibited lower mobility that PXN-EGFP- Y31F-EGFP-expressing CONCLUSIONS sustained steadfastness an incompletely active played pivotal role MMC-retarded