Connective tissue growth factor cooperates with fibronectin in enhancing attachment and migration of corneal epithelial cells.
Corneal wound healing is a complex process involving the integrated actions of various growth factors, cytokines and extracellular matrix produced by corneal cells and inflammatory cells. Connective tissue growth factor (CTGF) has been linked to wound healing, and fibronectin (FN) is a major component of the extracellular matrix. However, the functions of CTGF and FN in corneal epithelial cells are not well understood. We therefore investigated the coordinated function of CTGF and FN in the attachment and migration of corneal epithelial cells. Treatment of human corneal epithelial cells (HCECs) with transforming growth factor (TGF) beta1 up-regulated the expression of CTGF, but did not noticeably affect FN expression, as judged by immunoblot analysis of cell lysates. In contrast, the amount of FN accumulated in the cultured media was increased in a time-dependent manner, but CTGF was undetectable in the cultured media. The expression level of FN was decreased by the knockdown of CTGF expression with a specific short hairpin RNA, indicating that CTGF acts as an upstream mediator of FN expression. CTGF augmented the FN-mediated increase in the attachment of HCEC by about twofold, although CTGF alone did not influence the attachment. Moreover, the migration assay with rabbit corneal blocks revealed that CTGF (390 nM) alone or in combination of FN (10 microg/mL) promoted corneal epithelial migration; the mean migration distances of control, CTGF, and CTGF + FN were 272, 325, and 626, microm, respectively. In conclusion, CTGF cooperates with FN in enhancing the attachment and migration of corneal epithelial cells.
Sugioka
Koji
K
Department of Ophthalmology, Kinki University School of Medicine, Osaka-Sayama. sugioka@ganka.med.kindai.ac.jp
Yoshida
Koji
K
Kodama
Aya
A
Mishima
Hiroshi
H
Abe
Kosuke
K
Munakata
Hiroshi
H
Shimomura
Yoshikazu
Y
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001681:Biological Assay; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D055513:Connective Tissue Growth Factor / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology*; D005353:Fibronectins / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006801:Humans; D034741:RNA, Small Interfering / Q000378:metabolism; D011817:Rabbits; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
D000906:Antibodies; D005353:Fibronectins; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor