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K. Sugioka, A. Kodama, K. Yoshida, K. Abe, H. Mishima, Y. Shimomura; Interaction Between Connective Tissue Growth Factor and Fibronectin to Corneal Epithelial Cell Attachment and Migration. Invest. Ophthalmol. Vis. Sci. 2009;50(13):6310.
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© ARVO (1962-2015); The Authors (2016-present)
Recent studies have reported that connective tissue growth factor(CTGF) biochemically binds to fibronectin(FN) and promotes cell activity on various cells. In this study, we investigate the interaction between CTGF and FN for corneal epithelial cell attachment and migration.
A human corneal epithelial cells (HCEC) line that had been established by SV40-adenovirus recombinant vector was used in this study. CTGF-specific shRNA was synthesized and transfected into HCECs. The transfected cells were evaluated by western blot analysis to examine CTGF and FN production. For cell attachment assay, HCECs were either incubated with or without CTGF and were incubated in a FN-coated well or a BSA-coated well. After 45 minutes, the numbers of cells attached to the wells were counted. Another experiment was conducted to determine the interaction between CTGF and FN for epithelial cell migration. Rabbit cornea blocks were incubated with CTGF alone, FN alone or with a combination of both for 24hours and the length of migrated cells was measured.
At 48 hours after CTGF-specific shRNA transfection, both CTGF and FN productions in the HCECs reduced. In the HCECs incubated without CTGF, the number of cells that attached to the well significantly increased in the FN-coated well than in the BSA-coated well(p<0.01). The HCECs incubated with CTGF also showed a similar result with a significant increase in the number of cells that attached to the FN-coated wells(p<0.01). The result of migration assay using rabbit corneal organ culture indicated that CTGF promoted the corneal epithelial migration in a dose-dependent fashion in both samples with CTGF alone and with a combination of CTGF and FN.
CTGF and FN have cooperate action on the promotion of corneal epithelial attachment and migration.
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