Correlation of proliferative and anti-apoptotic effects of HGF, insulin, IGF-1, IGF-2, and EGF in SV40-transformed human corneal epithelial cells

The effects of various growth factors on the proliferation and apoptosis of human corneal epithelial cells were investigated. Simian virus 40-transformed human corneal epithelial cells were thus incubated separately with eight different growth factors, after which cell proliferation was evaluated by measurement of [3H]thymidine incorporation or with the MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay and apoptosis was quantified by the terminal deoxyribonucleotidyl transferase-mediated dUTP-biotin nick-end labeling assay. Phosphorylation of the protein kinase Akt, which plays an important role in anti-apoptotic signaling, was also assessed by immunoblot analysis. The growth factors examined could be classified into three groups on the basis of their effects on the proliferation and apoptosis of human corneal epithelial cells: hepatocyte growth factor (HGF), insulin, insulin-like growth factor (IGF)-1, IGF-2, and epidermal growth factor (EGF) each increased cell proliferation, inhibited the induction of apoptosis by sodium nitroprusside, and elicited the activation of Akt; transforming growth factor-β1 and -β2 inhibited [3H]thymidine incorporation but had no effect on sodium nitroprusside-induced apoptosis or on Akt activity; and platelet-derived growth factor-BB had no effects on the measured parameters. HGF, insulin, IGF-1, IGF-2, and EGF may thus contribute to maintenance of the corneal epithelium and coordinate the proliferative and apoptotic responses of this tissue.