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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Kay, Eunduck P. Lee, Jeong Goo Song, Jong-Suk Smith, Ronald E. |
| Description | Country affiliation: United States Author Affiliation: Lee JG ( Doheny Eye Institute, Keck School of Medicine of the University of Southern California, Los Angeles, California 90033, USA.) |
| Abstract | PURPOSE: FGF-2 stimulates cell proliferation of rabbit corneal endothelial cells (rCECs) by degrading the cyclin-dependent kinase inhibitor p27(Kip1) (p27) through its phosphorylation mechanism. The authors investigated whether the cell proliferation of human CECs (hCECs) is also induced by FGF-2 stimulation through the p27 phosphorylation pathway. METHODS: Expression and activation of protein were analyzed by immunoblotting. Cell proliferation was measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. Transfection of hCECs with small interference RNA (siRNA) was performed using a transfection reagent. RESULTS: FGF-2 stimulated cell proliferation in hCECs; the FGF-2 action was completely blocked by pathway-specific inhibitors for PI 3-kinase (LY294002) and MEK1/2 (U0126), respectively. Using immunoblotting, the authors showed that FGF-2 induced phosphorylation of p27 at both serine 10 (Ser10) and threonine 187 (Thr187) sites. These effects were also completely blocked by LY294002 or U0126. The authors then determined cross-talk between PI 3-kinase and extracellular signal-regulated kinase (ERK)1/2; blocking of ERK1/2 activation by LY294002 indicated that in hCECs ERK1/2 works as a downstream effector to PI 3-kinase for cell proliferation induced by FGF-2, whereas the ERK1/2 pathway in rCECs is parallel to the PI 3-kinase pathway. However, the downstream mechanism involved in cell cycle progression in hCECs is identical to that of rCECs: phosphorylation of p27 at Ser10 was mediated by kinase-interacting stathmin (KIS), confirmed with siRNA to KIS, and phosphorylation of p27 at Thr187 was mediated by cell division cycle 25A (Cdc25A), confirmed using Cdc25A inhibitor. CONCLUSIONS; FGF-2 stimulates proliferation of hCECs through PI 3-kinase and its downstream target ERK1/2 pathways. This linear signal transduction significantly downregulates p27 through its phosphorylation at both Ser10 and Thr187 sites mediated by KIS and Cdc25A, respectively. |
| ISSN | 01460404 |
| e-ISSN | 15525783 |
| DOI | 10.1167/iovs.11-8213 |
| Journal | Investigative Opthalmology & Visual Science |
| Issue Number | 11 |
| Volume Number | 52 |
| Language | English |
| Publisher | Association for Research in Vision and Ophthalmology |
| Publisher Date | 2011-10-17 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Cell Proliferation Drug Effects Cyclin-dependent Kinase Inhibitor P27 Metabolism Endothelium, Corneal Fibroblast Growth Factor 2 Pharmacology Phosphatidylinositol 3-kinases Adolescent Butadienes Chromones Enzyme Inhibitors Map Kinase Kinase 1 Antagonists & Inhibitors Map Kinase Kinase 2 Microscopy, Confocal Morpholines Nitriles Phosphorylation Rna, Small Interfering Genetics Serine Tetrazolium Salts Thiazoles Threonine Tissue Donors Transfection Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Discipline Ophthalmology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Ophthalmology Sensory Systems Cellular and Molecular Neuroscience |
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