Effects of epiplakin-knockdown in cultured corneal epithelial cells

BMC Res Notes. 2016 May 20:9:278. doi: 10.1186/s13104-016-2082-7.

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.

Keywords: Corneal epithelium; Culture; E-cadherin; Epiplakin; Knockdown; Migration; Proliferation.

MeSH terms

  • Autoantigens / genetics
  • Autoantigens / physiology*
  • Cadherins / metabolism
  • Cell Line, Transformed
  • Epithelium, Corneal / metabolism*
  • Gene Expression
  • Gene Knockdown Techniques
  • Humans
  • Keratin-6 / metabolism
  • Vimentin / metabolism

Substances

  • Autoantigens
  • Cadherins
  • Keratin-6
  • Vimentin
  • epiplakin