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(Investigative Ophthalmology and Visual Science. 2006;47:873-882.)
© 2006 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-0826
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Insulin-like Growth Factor-1 Induces Migration and Expression of Laminin-5 in Cultured Human Corneal Epithelial Cells

Hyung Keun Lee,1 Joon H. Lee,1 Min Kim,1 Yoshinobu Kariya,2 Kaoru Miyazaki,2 and Eung Kweon Kim1

1From the Department of Ophthalmology, Yonsei University College of Medicine, Seoul, Korea; and the 2Kihara Institute for Biological Research, Yokohama City University, Yokohama, Japan.

PURPOSE. The effects of insulin-like growth factor (IGF)-1 on laminin (Ln)-5 and the associated integrins during in vitro HCEC migration were examined. Also investigated were the effects of IGF-1 on the migration of human corneal epithelial cells (HCECs).

METHODS. HCEC migration was examined by wound-healing and chemoattraction assays. For migration inhibition assays, HCECs were pretreated with inhibitors of the IGF-1 receptor ({alpha}IR3), the PI3-K/AKT pathway (LY294002), and the MEK-ERK pathway (PD98059). The expression levels of Ln-5 and fibronectin (Fn) were determined by Western blot analysis, and the expression levels of the ß1 and {alpha}3 integrins were determined by confocal microscopy and Western blot analysis. The migration inhibition with anti-integrin {alpha}3 and ß1 antibodies was also determined.

RESULTS. HCEC migration was significantly increased in the presence of IGF-1 and Ln-5. IGF-1 enhanced the production of Ln-5 in both a dose- and time-dependent manner, and this upregulation was blocked by pretreatment with {alpha}IR3 or LY294002. IGF-1 treatment upregulated expression of ß1 integrin, but not {alpha}3 integrin. The HCEC migration facilitated by IGF-1 was inhibited with the anti-integrin antibody for ß1. However, there was no cross-talk between Ln-5 and integrin ß1 production.

CONCLUSIONS. The results reveal that IGF-1 induces HCEC migration through the independent production of Ln-5 and ß1 integrin, which are directed at least in part by activation of the PI3-K/AKT pathway, but are not affected by the MEK-ERK pathway.




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