title: Subepithelial corneal fibrosis partially due to epithelial-mesenchymal transition of ocular surface epithelium.
abstract: OBJECTIVE
  To determine whether epithelial-mesenchymal transition is involved in the development of corneal subepithelial fibrosis (pannus).
  
  
  METHODS
  Frozen samples of pannus tissue removed from human corneas with a diagnosis of total limbal stem cell deficiency were characterized by immunostaining for both epithelial and mesenchymal markers. We selected transformation-related protein 63 (p63) and pancytokeratin as epithelial markers and vimentin and α-smooth muscle actin (α-SMA) as mesenchymal markers. Immunostaining for β-catenin and E-cadherin was performed to determine wingless-Int (Wnt)-pathway activation. RT-PCR analysis was also performed on epithelial tissue obtained from pannus samples after dispase digestion.
  
  
  RESULTS
  Immunohistochemistry revealed strong nuclear expression of p63 and weak intercellular expression of E-cadherin in epithelial basal cells of pannus tissue. Furthermore, translocation of β-catenin from intercellular junctions to the nucleus and cytoplasm was also observed. Double-positive cells for both p63 and α-SMA were observed in the subepithelial stroma of pannus tissue, which was supported by RT-PCR and cytospin analysis.
  
  
  CONCLUSIONS
  Epithelial-mesenchymal transition may be partially involved in the development of subepithelial corneal fibrosis due to total limbal stem cell deficiency.
authors.affiliation: Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Motoko
authors.initials: M
authors.lastname: Kawashima
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kawakita
authors.forename: Kazunari
authors.initials: K
authors.lastname: Higa
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Satake
authors.forename: Masahiro
authors.initials: M
authors.lastname: Omoto
authors.forename: Kazuo
authors.initials: K
authors.lastname: Tsubota
authors.forename: Shigeto
authors.initials: S
authors.lastname: Shimmura
authors.forename: Jun
authors.initials: J
authors.lastname: Shimazaki
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015820:Cadherins / Q000378:metabolism; D002648:Child; D003710:Demography; D058750:Epithelial-Mesenchymal Transition; D019573:Epithelium, Corneal / Q000201:enzymology / Q000473:pathology*; D005260:Female; D005355:Fibrosis; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008875:Middle Aged; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism; D051176:beta Catenin / Q000378:metabolism
chemical_list: D000199:Actins; D015820:Cadherins; C115051:TP63 protein, human; D015534:Trans-Activators; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D014746:Vimentin; D051176:beta Catenin