Role of HGF/c-Met in serum-starved ARPE-19 cells.
OBJECTIVE
Hepatocyte growth factor (HGF) and its receptor (HGFR/c-Met) regulate motility, mitogenesis, and morphogenesis in a cell type-dependent fashion. We report the role of HGF and c-Met on stress-induced ARPE-19 human retinal pigment epithelial (RPE) cells in this study.


METHODS
The cells were cultured either with or without serum. Southern and Western blot analyses were done to determine the expression patterns of HGF/c-Met in serum-starved ARPE-19 cells. The cell proliferation pattern in serum-starved condition was analyzed using MTS assay. Inhibition level of cell proliferation was analyzed using a neutralizing monoclonal antibody against c-Met (2 microg/ml).


RESULTS
Abnormal cell proliferation and scattering of ARPE-19 cells was observed under serum starvation. HGF/c-Met were expressed in serum-starved ARPE-19 cells. ARPE-19 cell proliferation was also enhanced with recombinant HGF treatment. Neutralization against c-Met inhibited the proliferation of serum-deprived ARPE-19 by 64.5% (n=9, S.D. 5.5%). Serum starvation appears to induce epithelial-mesenchymal transition of ARPE-19 cells, resulting in scatter, and the expression of alpha-smooth muscle actin (alpha-SMA), a marker for fibrosis.


CONCLUSIONS
In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.
Department of Ophthalmology, College of Medicine, The Catholic University of Korea, Seoul, Korea.
Eun Jung
EJ
Jun
Hwa Sun
HS
Kim
Yeong Hoon
YH
Kim
D015139:Blotting, Southern; D015153:Blotting, Western; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D015870:Gene Expression; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008938:Mitosis / Q000502:physiology; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D019859:Proto-Oncogene Proteins c-met / Q000096:biosynthesis / Q000235:genetics*; D012313:RNA / Q000235:genetics*
D016895:Culture Media, Serum-Free; D012313:RNA; D017228:Hepatocyte Growth Factor; D019859:Proto-Oncogene Proteins c-met