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A. Kodama, K. Sugioka, H. Mishima, K. Yoshida, K. Abe, Y. Shimomura; Interaction Between Connective Tissue Growth Factor and Fibronectin in Attachment and Migration of Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1952.
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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 in corneal epithelial cell attachment and migration.
A human corneal epithelial cell (HCEC) line that had been established by a SV40-adenovirus recombinant vector was used in this study. CTGF-specific short hairpin RNA (shRNA) was synthesized and transfected into HCECs. The transfected cells were evaluated by western blot analysis to examine CTGF and FN production. For the cell attachment assay, HCECs were incubated with CTGF, FN, or CTGF+FN. HCECs were also incubated with or without CTGF in FN-coated wells or bovine serum albumin (BSA)-coated wells. After 45 minutes, the number of cells attached to the wells was counted. Another experiment was conducted to determine the interaction between CTGF and FN in epithelial cell migration. Rabbit cornea blocks were incubated with CTGF alone, FN alone, or with a combination of both for 24 hours and the distance of cell migration was measured.
At 48 hours after CTGF-specific shRNA transfection, CTGF and FN production in the HCECs was reduced. For the cell attachment assay, CTGF increased the number of cells attached in the presence of FN, whereas CTGF alone did not increase the number of cells attached (p<0.01). Moreover, anti-FN antibody completely inhibited CTGF-stimulated cellular adhesion to FN (p<0.05). The result of the migration assay using rabbit corneal organ culture indicated that CTGF promoted corneal epithelial migration in a dose-dependent fashion in samples with CTGF alone or with a combination of CTGF and FN.
CTGF and FN have cooperative action in promoting corneal epithelial attachment and migration
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