MAP kinase and β-catenin signaling in HGF induced RPE migration

Gregory I. Liou, Suraporn Matragoon, Sara Samuel, M. Ali Behzadian, Nai Tse Tsai, Xiaolin Gu, Penny Roon, D. Margaret Hunt, Richard C. Hunt, Ruth B Caldwell, Dennis M. Marcus

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Purpose: Hepatocyte growth factor (HGF) has been implicated in retinal pigment epithelial (RPE) cell proliferation and migration that occurs in proliferative retinal diseases such as proliferative vitreoretinopathy (PVR). The aim of this study is to investigate HGF induced signaling pathways that lead to RPE cell migration. Methods: Localization of β-catenin was determined by immunofluorescence. HGF induced migration of ARPE-19 cells was studied using a quantitative migration assay after wounding in the presence of a DNA polymerase inhibitor, and in the presence or absence of a mitogen activated protein kinase (MAP kinase) kinase inhibitor. C-jun expression was determined by semi-quantitative RT-PCR and by Northern blot analysis. P42/p44 MAP kinase activity was determined by western blot and by an immunoprecipitation kinase assay. Tyrosine phosphorylation of the HGF receptor (HGFR or c-met) and β-catenin was determined by immunoprecipitation and western blot analysis. Transactivation activity of β-catenin was determined by luciferase reporter gene analysis. Results: β-catenin and E-cadherin were co-localized on the basal surface of the RPE in vivo. Diffusion of the cell surface-localized β-catenin occurs in migratory cells in vitro in the presence of HGF. HGF induced a MAP kinase dependent ARPE-19 cell migration, which is accompanied with a transient increase of c-jun expression and concomitant increases of MAP kinase activity, tyrosine phosphorylation of HGFR and β-catenin, increased cytosolic levels of β-catenin, and transactivation activity of β-catenin. Tyrosine phosphorylation of HGFR and β-catenin occurs in the primary or passaged RPE cultures or proliferative ARPE-19 cells, but not freshly isolated RPE or differentiated ARPE-19 cells. Conclusions: This study defines the signal transduction pathways activated by HGF in RPE cells, leading to an increase in the MAP kinase activity and free pool of β-catenin, and changes in gene expression. These findings are consistent with the hypothesis that both β-catenin and MAP kinases are components of the HGF induced RPE migration that occurs in proliferative retinal diseases.

Original languageEnglish (US)
Pages (from-to)483-493
Number of pages11
JournalMolecular Vision
Volume8
StatePublished - Dec 1 2002

Fingerprint

Catenins
Retinal Pigments
Hepatocyte Growth Factor
Mitogen-Activated Protein Kinases
Cell Movement
Tyrosine
Retinal Diseases
Epithelial Cells
Phosphorylation
Immunoprecipitation
Transcriptional Activation
Phosphotransferases
Western Blotting
Proto-Oncogene Proteins c-met
Proliferative Vitreoretinopathy
Nucleic Acid Synthesis Inhibitors
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase Kinases
Cadherins
Luciferases

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Liou, G. I., Matragoon, S., Samuel, S., Behzadian, M. A., Tsai, N. T., Gu, X., ... Marcus, D. M. (2002). MAP kinase and β-catenin signaling in HGF induced RPE migration. Molecular Vision, 8, 483-493.

MAP kinase and β-catenin signaling in HGF induced RPE migration. / Liou, Gregory I.; Matragoon, Suraporn; Samuel, Sara; Behzadian, M. Ali; Tsai, Nai Tse; Gu, Xiaolin; Roon, Penny; Hunt, D. Margaret; Hunt, Richard C.; Caldwell, Ruth B; Marcus, Dennis M.

In: Molecular Vision, Vol. 8, 01.12.2002, p. 483-493.

Research output: Contribution to journalArticle

Liou, GI, Matragoon, S, Samuel, S, Behzadian, MA, Tsai, NT, Gu, X, Roon, P, Hunt, DM, Hunt, RC, Caldwell, RB & Marcus, DM 2002, 'MAP kinase and β-catenin signaling in HGF induced RPE migration', Molecular Vision, vol. 8, pp. 483-493.
Liou GI, Matragoon S, Samuel S, Behzadian MA, Tsai NT, Gu X et al. MAP kinase and β-catenin signaling in HGF induced RPE migration. Molecular Vision. 2002 Dec 1;8:483-493.
Liou, Gregory I. ; Matragoon, Suraporn ; Samuel, Sara ; Behzadian, M. Ali ; Tsai, Nai Tse ; Gu, Xiaolin ; Roon, Penny ; Hunt, D. Margaret ; Hunt, Richard C. ; Caldwell, Ruth B ; Marcus, Dennis M. / MAP kinase and β-catenin signaling in HGF induced RPE migration. In: Molecular Vision. 2002 ; Vol. 8. pp. 483-493.
@article{908764986ff648749f984d75349aa0ab,
title = "MAP kinase and β-catenin signaling in HGF induced RPE migration",
abstract = "Purpose: Hepatocyte growth factor (HGF) has been implicated in retinal pigment epithelial (RPE) cell proliferation and migration that occurs in proliferative retinal diseases such as proliferative vitreoretinopathy (PVR). The aim of this study is to investigate HGF induced signaling pathways that lead to RPE cell migration. Methods: Localization of β-catenin was determined by immunofluorescence. HGF induced migration of ARPE-19 cells was studied using a quantitative migration assay after wounding in the presence of a DNA polymerase inhibitor, and in the presence or absence of a mitogen activated protein kinase (MAP kinase) kinase inhibitor. C-jun expression was determined by semi-quantitative RT-PCR and by Northern blot analysis. P42/p44 MAP kinase activity was determined by western blot and by an immunoprecipitation kinase assay. Tyrosine phosphorylation of the HGF receptor (HGFR or c-met) and β-catenin was determined by immunoprecipitation and western blot analysis. Transactivation activity of β-catenin was determined by luciferase reporter gene analysis. Results: β-catenin and E-cadherin were co-localized on the basal surface of the RPE in vivo. Diffusion of the cell surface-localized β-catenin occurs in migratory cells in vitro in the presence of HGF. HGF induced a MAP kinase dependent ARPE-19 cell migration, which is accompanied with a transient increase of c-jun expression and concomitant increases of MAP kinase activity, tyrosine phosphorylation of HGFR and β-catenin, increased cytosolic levels of β-catenin, and transactivation activity of β-catenin. Tyrosine phosphorylation of HGFR and β-catenin occurs in the primary or passaged RPE cultures or proliferative ARPE-19 cells, but not freshly isolated RPE or differentiated ARPE-19 cells. Conclusions: This study defines the signal transduction pathways activated by HGF in RPE cells, leading to an increase in the MAP kinase activity and free pool of β-catenin, and changes in gene expression. These findings are consistent with the hypothesis that both β-catenin and MAP kinases are components of the HGF induced RPE migration that occurs in proliferative retinal diseases.",
author = "Liou, {Gregory I.} and Suraporn Matragoon and Sara Samuel and Behzadian, {M. Ali} and Tsai, {Nai Tse} and Xiaolin Gu and Penny Roon and Hunt, {D. Margaret} and Hunt, {Richard C.} and Caldwell, {Ruth B} and Marcus, {Dennis M.}",
year = "2002",
month = "12",
day = "1",
language = "English (US)",
volume = "8",
pages = "483--493",
journal = "Molecular Vision",
issn = "1090-0535",

}

TY - JOUR

T1 - MAP kinase and β-catenin signaling in HGF induced RPE migration

AU - Liou, Gregory I.

AU - Matragoon, Suraporn

AU - Samuel, Sara

AU - Behzadian, M. Ali

AU - Tsai, Nai Tse

AU - Gu, Xiaolin

AU - Roon, Penny

AU - Hunt, D. Margaret

AU - Hunt, Richard C.

AU - Caldwell, Ruth B

AU - Marcus, Dennis M.

PY - 2002/12/1

Y1 - 2002/12/1

N2 - Purpose: Hepatocyte growth factor (HGF) has been implicated in retinal pigment epithelial (RPE) cell proliferation and migration that occurs in proliferative retinal diseases such as proliferative vitreoretinopathy (PVR). The aim of this study is to investigate HGF induced signaling pathways that lead to RPE cell migration. Methods: Localization of β-catenin was determined by immunofluorescence. HGF induced migration of ARPE-19 cells was studied using a quantitative migration assay after wounding in the presence of a DNA polymerase inhibitor, and in the presence or absence of a mitogen activated protein kinase (MAP kinase) kinase inhibitor. C-jun expression was determined by semi-quantitative RT-PCR and by Northern blot analysis. P42/p44 MAP kinase activity was determined by western blot and by an immunoprecipitation kinase assay. Tyrosine phosphorylation of the HGF receptor (HGFR or c-met) and β-catenin was determined by immunoprecipitation and western blot analysis. Transactivation activity of β-catenin was determined by luciferase reporter gene analysis. Results: β-catenin and E-cadherin were co-localized on the basal surface of the RPE in vivo. Diffusion of the cell surface-localized β-catenin occurs in migratory cells in vitro in the presence of HGF. HGF induced a MAP kinase dependent ARPE-19 cell migration, which is accompanied with a transient increase of c-jun expression and concomitant increases of MAP kinase activity, tyrosine phosphorylation of HGFR and β-catenin, increased cytosolic levels of β-catenin, and transactivation activity of β-catenin. Tyrosine phosphorylation of HGFR and β-catenin occurs in the primary or passaged RPE cultures or proliferative ARPE-19 cells, but not freshly isolated RPE or differentiated ARPE-19 cells. Conclusions: This study defines the signal transduction pathways activated by HGF in RPE cells, leading to an increase in the MAP kinase activity and free pool of β-catenin, and changes in gene expression. These findings are consistent with the hypothesis that both β-catenin and MAP kinases are components of the HGF induced RPE migration that occurs in proliferative retinal diseases.

AB - Purpose: Hepatocyte growth factor (HGF) has been implicated in retinal pigment epithelial (RPE) cell proliferation and migration that occurs in proliferative retinal diseases such as proliferative vitreoretinopathy (PVR). The aim of this study is to investigate HGF induced signaling pathways that lead to RPE cell migration. Methods: Localization of β-catenin was determined by immunofluorescence. HGF induced migration of ARPE-19 cells was studied using a quantitative migration assay after wounding in the presence of a DNA polymerase inhibitor, and in the presence or absence of a mitogen activated protein kinase (MAP kinase) kinase inhibitor. C-jun expression was determined by semi-quantitative RT-PCR and by Northern blot analysis. P42/p44 MAP kinase activity was determined by western blot and by an immunoprecipitation kinase assay. Tyrosine phosphorylation of the HGF receptor (HGFR or c-met) and β-catenin was determined by immunoprecipitation and western blot analysis. Transactivation activity of β-catenin was determined by luciferase reporter gene analysis. Results: β-catenin and E-cadherin were co-localized on the basal surface of the RPE in vivo. Diffusion of the cell surface-localized β-catenin occurs in migratory cells in vitro in the presence of HGF. HGF induced a MAP kinase dependent ARPE-19 cell migration, which is accompanied with a transient increase of c-jun expression and concomitant increases of MAP kinase activity, tyrosine phosphorylation of HGFR and β-catenin, increased cytosolic levels of β-catenin, and transactivation activity of β-catenin. Tyrosine phosphorylation of HGFR and β-catenin occurs in the primary or passaged RPE cultures or proliferative ARPE-19 cells, but not freshly isolated RPE or differentiated ARPE-19 cells. Conclusions: This study defines the signal transduction pathways activated by HGF in RPE cells, leading to an increase in the MAP kinase activity and free pool of β-catenin, and changes in gene expression. These findings are consistent with the hypothesis that both β-catenin and MAP kinases are components of the HGF induced RPE migration that occurs in proliferative retinal diseases.

UR - http://www.scopus.com/inward/record.url?scp=0036978987&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036978987&partnerID=8YFLogxK

M3 - Article

C2 - 12500177

AN - SCOPUS:0036978987

VL - 8

SP - 483

EP - 493

JO - Molecular Vision

JF - Molecular Vision

SN - 1090-0535

ER -