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Role of HGF/c-Met in serum-starved ARPE-19 cells.

Jun EJ, Kim HS, Kim YH - Korean J Ophthalmol (2007)

Bottom Line: 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%).In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, College of Medicine, The Catholic University of Korea, Seoul, Korea.

ABSTRACT

Purpose: 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.

Show MeSH

Related in: MedlinePlus

ARPE-19 cell proliferation patterns and scattering during serum-starvation. Serum-starved ARPE-19 cells were counted using a hemocytometer (A). MTS assay was performed at 490 nm on a micro-enzyme-linked immunosorbent assay (ELISA) plate reader (B). ARPE-19 cells showed scattering pattern and spindle shape after serum starvation for 24 hours (C) and (D).
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Figure 1: ARPE-19 cell proliferation patterns and scattering during serum-starvation. Serum-starved ARPE-19 cells were counted using a hemocytometer (A). MTS assay was performed at 490 nm on a micro-enzyme-linked immunosorbent assay (ELISA) plate reader (B). ARPE-19 cells showed scattering pattern and spindle shape after serum starvation for 24 hours (C) and (D).

Mentions: The growth rate of serum-starved RPE cells was equal or increased compared to that of ARPE-19 cells in normal culture conditions (Fig. 1A and B). The cells were scattered and spindle-like in shape (Fig. 1C and D) after starvation.


Role of HGF/c-Met in serum-starved ARPE-19 cells.

Jun EJ, Kim HS, Kim YH - Korean J Ophthalmol (2007)

ARPE-19 cell proliferation patterns and scattering during serum-starvation. Serum-starved ARPE-19 cells were counted using a hemocytometer (A). MTS assay was performed at 490 nm on a micro-enzyme-linked immunosorbent assay (ELISA) plate reader (B). ARPE-19 cells showed scattering pattern and spindle shape after serum starvation for 24 hours (C) and (D).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2629891&req=5

Figure 1: ARPE-19 cell proliferation patterns and scattering during serum-starvation. Serum-starved ARPE-19 cells were counted using a hemocytometer (A). MTS assay was performed at 490 nm on a micro-enzyme-linked immunosorbent assay (ELISA) plate reader (B). ARPE-19 cells showed scattering pattern and spindle shape after serum starvation for 24 hours (C) and (D).
Mentions: The growth rate of serum-starved RPE cells was equal or increased compared to that of ARPE-19 cells in normal culture conditions (Fig. 1A and B). The cells were scattered and spindle-like in shape (Fig. 1C and D) after starvation.

Bottom Line: 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%).In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, College of Medicine, The Catholic University of Korea, Seoul, Korea.

ABSTRACT

Purpose: 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.

Show MeSH
Related in: MedlinePlus