2. Academic Validation
  3. Effects of epiplakin-knockdown in cultured corneal epithelial cells

Effects of epiplakin-knockdown in cultured corneal epithelial cells

  • BMC Res Notes. 2016 May 20;9:278. doi: 10.1186/s13104-016-2082-7.
Masahide Kokado 1,  Yuka Okada 2,  Takeshi Miyamoto 2,  Osamu Yamanaka 2,  Shizuya Saika 2
Affiliations

Affiliations

  • 1 Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan. [email protected].
  • 2 Department of Ophthalmology, Wakayama Medical University School of Medicine, 811-1 Kimiidera, Wakayama, 641-0012, Japan.
PMID: 27206504 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.

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