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J.-S. Song, J. Lee, R. E. Smith, E. P. Kay; In vitro and ex vivo Effects of FGF-2 on Cell Proliferation and p27 Expression in Human Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1812.
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Fibroblast growth factor 2 (FGF-2) is known to stimulate cell proliferation of rabbit corneal endothelial cells (CECs) by degrading p27. We investigated the effect of FGF-2 on cell proliferation and p27 expression in human CECs in ex vivo and in vitro studies.
hCECs were isolated with EDTA treatment; cell proliferation was determined by MTT assay. Pharmacological inhibitors were used to block PI 3-kinase and ERK1/2. Subcellular localization of p27 and phosphorylated p27 (pp27) was determined using immunofluorescent staining.
: FGF-2 stimulated cell proliferation in hCECs and these effects were inhibited by LY294002 and U0126. Cells maintained in mitogen-deprived medium were stained for p27 in the nuclei, but not for pp27. However, FGF-2 abolished the staining of p27, while nuclear staining of pp27 was observed. Such effect of FGF-2 on p27 and pp27 expression was blocked with LY294002. Likewise, the ex vivo human corneal endothelium showed nuclear p27 staining in mitogen-deprived medium. However, p27 expression was significantly decreased with FGF-2 treatment.
FGF-2 stimulates cell proliferation in hCECs through PI-3 kinase and ERK 1/2 pathways and significantly downregulates p27 through phosphorylation mechanism, as observed in rabbit CECs.
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