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Mitogenic and antiapoptotic effects of various growth factors on human corneal fibroblasts
Ryoji Yanai1, Naoyuki Yamada, Naruji Kugimiya
1Department of Biomolecular Recognition and Ophthalmology, Yamaguchi University School of Medicine, 1-1-1 Minami-Kogushi, Ube City, Yamaguchi 755-8505, Japan.
Purpose:
To investigate both the effects of various growth factors on the proliferation of human corneal fibroblasts and the abilities of these factors to protect the cells from apoptosis.
Methods:
Cultured human corneal fibroblasts were incubated separately with 11 different growth factors whose receptors are expressed by these cells. Cell proliferation was evaluated by measurement of [(3)H]thymidine incorporation. The activation of the protein kinase Akt, which plays an important role in antiapoptotic signaling, was assessed by immunoblot analysis with antibodies specific for a phosphorylated form of the enzyme. Apoptosis was quantitated by the TdT-mediated dUTP-biotin nick-end labeling (TUNEL) assay.
Results:
Of the 11 growth factors examined, platelet-derived growth factor, insulin, insulin-like growth factors-1 and -2, and epidermal growth factor, each stimulated the proliferation of corneal fibroblasts, induced the activation of Akt in these cells, and protected them from apoptosis induced by sodium nitroprusside (SNP). Basic fibroblast growth factor, keratinocyte growth factor, nerve growth factor, and hepatocyte growth factor stimulated cell proliferation but did not induce Akt activation or protect the cells from SNP-induced apoptosis. Transforming growth factor-beta1 and -beta2 did not stimulate proliferation and had no effect on Akt activity or on SNP-induced apoptosis.
Conclusions:
In terms of their modulatory effects on the proliferation and apoptosis of human corneal fibroblasts, the 11 growth factors examined can be classified into three groups. These growth factors may both contribute to maintenance of the cornea and coordinate the proliferative and apoptotic responses of corneal fibroblasts during corneal wound healing.
Insights
Certain growth factors promote human corneal fibroblast proliferation and survival by activating Akt signaling. These findings classify growth factors into three groups, aiding understanding of corneal wound healing and maintenance.
Area of Science:
- Ophthalmology
- Cell Biology
- Biochemistry
Background:
- Human corneal fibroblasts are crucial for maintaining corneal integrity.
- Understanding growth factor roles in fibroblast proliferation and apoptosis is key for corneal wound healing research.
Purpose of the Study:
- To investigate the effects of 11 growth factors on human corneal fibroblast proliferation.
- To determine the ability of these growth factors to protect corneal fibroblasts from apoptosis.
Main Methods:
- Human corneal fibroblasts were cultured and treated with 11 growth factors.
- Cell proliferation was measured using [(3)H]thymidine incorporation.
- Akt activation and apoptosis were assessed via immunoblot analysis and TUNEL assay, respectively.
Main Results:
- Platelet-derived growth factor, insulin, IGF-1, IGF-2, and EGF stimulated proliferation, activated Akt, and protected cells from apoptosis.
- bFGF, KGF, NGF, and HGF stimulated proliferation but did not affect Akt or apoptosis.
- TGF-β1 and TGF-β2 had no effect on proliferation, Akt activity, or apoptosis.
Conclusions:
- Growth factors differentially modulate human corneal fibroblast proliferation and apoptosis.
- These factors can be categorized into three distinct functional groups.
- Findings may inform corneal maintenance and wound healing strategies.
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