title: Effects of epiplakin-knockdown in cultured corneal epithelial cells.
abstract: BACKGROUND
  To investigate effects of knockdown of epiplakin gene expression on the homeostasis of cultured corneal epithelial cell line. We previously reported acceleration of corneal epithelial wound healing in an epiplakin-null mouse.
  
  
  METHODS
  Gene expression of epiplakin was knockdowned by employing siRNA transfection in SV40-immortalized human corneal epithelial cell line. Protein expression of E-cadherin, keratin 6 and vimentin was examined by western blotting. Cell migration and proliferation were examined by using scratch assay and Alamar blue assay, respectively.
  
  
  RESULTS
  Scratch assay and Alamar blue assay showed migration and proliferation of the cells was accelerated by epiplakin knockdown. siRNA-knockdown of epiplakin suppressed protein expression of E-cadherin, keratin 6 and vimentin.
  
  
  CONCLUSIONS
  Decreased expression of E-cadherin, keratin 6 and vimentin might be included in the mechanisms of cell migration acceleration in the absence of epiplakin. The mechanism of cell proliferation stimulation by epiplakin knockdown is to be investigated.
authors.affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan. kokado@wakayama-med.ac.jp.
authors.forename: Masahide
authors.initials: M
authors.lastname: Kokado
authors.affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Okada
authors.affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Miyamoto
authors.affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Yamanaka
authors.affiliation: Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan.
authors.forename: Shizuya
authors.initials: S
authors.lastname: Saika
mesh_terms: D001324:Autoantigens / Q000235:genetics / Q000502:physiology*; D015820:Cadherins / Q000378:metabolism; D002461:Cell Line, Transformed; D019573:Epithelium, Corneal / Q000378:metabolism*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D006801:Humans; D053553:Keratin-6 / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
chemical_list: D001324:Autoantigens; D015820:Cadherins; D053553:Keratin-6; D014746:Vimentin; C426678:epiplakin
keywords: Corneal epithelium; Culture; E-cadherin; Epiplakin; Knockdown; Migration; Proliferation