title: Molecular mechanism of fluoroquinolones modulation on corneal fibroblast motility.
abstract: Topical fluoroquinolones are widely used to prevent ocular infections after ophthalmic surgery. However, they have been shown to affect the corneal cell motility, whose mechanism remains indefinite. The purpose of this study was to investigate how fluoroquinolones affect corneal stromal cell motility. Human corneal fibroblasts (HCFs) were incubated in ciprofloxacin (CIP), levofloxacin (LEV), or moxifloxacin (MOX) at 0, 10, 50, and 100 μg/ml for up to 3 days. Effect of CIP, LEV, or MOX on HCF migration was monitored using migration assay. HCF viability was determined by WST-1 assay. Expression of focal adhesion kinase (FAK), paxillin (PXN), and their phosphorylated forms were analyzed by immunoblotting. Binding affinity between FAK and PXN was determined by co-immunoprecipitation. Our results revealed that CIP and MOX, but not LEV, noticeably retarded HCF migration. HCF proliferation was significantly reduced by CIP (38.2%), LEV (29.5%), and MOX (21.3%), respectively (p = 0.002). CIP and MOX suppressed the phosphorylation of PXN at tyrosines (10.2 ± 4.3%, p < 0.001; 11.7 ± 2.4%, p < 0.001, respectively), including tyrosine 118 (33.3 ± 5.2%, p < 0.001; 34.0 ± 4.4%, p < 0.001, respectively). CIP and MOX diminished the binding affinity between FAK and PXN (8.2 ± 1.8%, p < 0.001; 9.0 ± 4.5%, p < 0.001, respectively). Nevertheless, tyrosine dephosphorylation and FAK dissociation of PXN were not found in LEV-treated HCFs. None of these fluoroquinolones affect phosphorylation of FAK-Y397. We conclude that CIP and MOX, but not LEV, might delay corneal fibroblast migration via interfering with recruitment of PXN to focal adhesions and dephosphorylation of PXN at the tyrosines.
authors.affiliation: Department of Ophthalmology, Far Eastern Memorial Hospital, New Taipei City, Taiwan.
authors.forename: Tsan-Chi
authors.initials: TC
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology, Far Eastern Memorial Hospital, New Taipei City, Taiwan; Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan.
authors.forename: Tzu-Yun
authors.initials: TY
authors.lastname: Tsai
authors.affiliation: Department of Ophthalmology, Far Eastern Memorial Hospital, New Taipei City, Taiwan; Department of Ophthalmology, National Taiwan University Hospital, Taipei, Taiwan. Electronic address: swchang2007@ntu.edu.tw.
authors.forename: Shu-Wen
authors.initials: SW
authors.lastname: Chang
mesh_terms: D000328:Adult; D000368:Aged; D000900:Anti-Bacterial Agents / Q000008:administration & dosage; D002452:Cell Count; D002465:Cell Movement / Q000187:drug effects; D003320:Corneal Ulcer / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D019573:Epithelium, Corneal / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D024841:Fluoroquinolones / Q000008:administration & dosage*; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000096:biosynthesis* / Q000187:drug effects; D006801:Humans; D015151:Immunoblotting; D008297:Male; D008875:Middle Aged; D009883:Ophthalmic Solutions; D051419:Paxillin / Q000096:biosynthesis* / Q000187:drug effects; D055815:Young Adult
chemical_list: D000900:Anti-Bacterial Agents; D024841:Fluoroquinolones; D009883:Ophthalmic Solutions; D051419:Paxillin; D051416:Focal Adhesion Protein-Tyrosine Kinases
keywords: Cell motility; Ciprofloxacin; FAK; Fluoroquinolone; Human corneal fibroblasts; Levofloxacin; Moxifloxacin; Paxillin