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K. Kimura, K. Kawamoto, S. Teranishi, K. Fukuda, A. Hattori, T. Nishida; Promotion of Corneal Epithelial Cell Migration by PHSRN, a Peptide Corresponding to a Cell Binding Domain of Fibronectin . Invest. Ophthalmol. Vis. Sci. 2005;46(13):2107.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Fibronectin plays an important role in the migration of corneal epithelial cells. The RGD sequence in the cell binding site of fibronectin mediates the interaction between fibronectin and its integrin receptor, whereas the PHSRN sequence, which constitutes another cell binding domain of fibronectin, contributes to cell adhesion. We examined the possible effect of a synthetic PHSRN peptide on corneal epithelial cell migration in vitro Methods: Simian virus 40–transformed human corneal epithelial (HCE) cells were cultured, harvested by exposure to trypsin–EDTA, and replated in 96–well plates in the absence or presence of PHSRN or the control peptide NRSHP at concentrations of 0.2 nM to 200 µM. The number of attached cells was counted after incubation for 1 hour. The effect of PHSRN on HCE cell proliferation in vitro was assessed by monitoring of mitochondrial metabolic activity with the MTS substrate. The behavior of cells plated on glass–bottom dishes with or without PHSRN or NRSHP was monitored by video microscopy. Cell migration was evaluated with a transwell assay. Results: The PHSRN peptide had no effect on the adhesion or proliferation of HCE cells. In contrast, PHSRN, but not NRSHP, promoted HCE cell migration in the transwell assay in a concentration–dependent manner. Video microscopy also revealed that PHSRN induced ruffling at the cell periphery and promoted cell migration, whereas NRSHP had no such effects. Conclusions: These results demonstrate that the fibronectin–derived PHSRN peptide stimulates the migration of HCE cells in vitro.
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