J Korean Ophthalmol Soc.  2001 Sep;42(9):1336-1343.

The Effect of Hydrogen Peroxide on the Migration and FAK Activation in Lens Epithelial Cells

• Park YM
• Kim JA
• Ahn BT
• Wee WR
• Lee JH
• Lee YC
• Kim HS
• Joo CK

Abstract

PURPOSE: Hydrogen peroxide has been implicated as a causative factor of various cellular dysfunction, cell death, transformation. In addition, oxidative stress has been suggested as a crucial inducer of cataract formation not only the nuclear-type cataract but also anterior polar type cataract. Transformation of lens epithelial cells accompanying accumulation of extracellular matrix molecules, and cell migration is observed in the cataract forming area of lens. In the present study, we investigated in the migration of lens epithelial cells and the activation of focal adhesion kinase(FAK), because there have been many reports showing that activation of FAK increased cell migration. METHOD: After treatment hygrogen peroxide in a dose-dependent on HLE B-3 cell line, we have performed immuno fluorenscence staing of actin stress fiber and migration assay, and then isolated total RNA to identify expression of MMP-2 from the cell line. To examine FAK activity, we are performed Phosphotyrosine Immunoblot analysis. RESULT: We observed the increased cell migration in response to hydrogen peroxide in a dose dependent manner. In addition, we observed that the phosphorylation of focal adhesion kinase was increased with the treatment of hydrogen peroxide. Also, we examined the expression of the matrix metalloproteinases-2(MMP-2) which disintegrates extracellular matrix and participates in cell migration. The result showed that the mRNA level of MMP-2 did not increase by hydrogen peroxide.
CONCLUSIONS
The results suggest that hydrogen peroxide enhanced migration of lens epithelial cell, and that FAK may play a role in the process of cell migration. In conclusion, migration of lens epithelial cell and phosphorylation of focal adhesion kinase was increased by treatment of hydrogen peroxide. Thus oxidative stress plays a crucial role in the transformation of lens epithelial cells and anterior polar type cataract formation.

Keyword

Focal adhesion Kinase; Lens epithelial cells; Migration; Oxidative stress

MeSH Terms

Actins
Cataract
Cell Death
Cell Line
Cell Movement
Epithelial Cells*
Extracellular Matrix
Focal Adhesion Protein-Tyrosine Kinases
Focal Adhesions
Hydrogen Peroxide*
Hydrogen*
Oxidative Stress
Phosphorylation
Phosphotyrosine
RNA
RNA, Messenger
Stress Fibers
Actins
Focal Adhesion Protein-Tyrosine Kinases
Hydrogen
Hydrogen Peroxide
Phosphotyrosine
RNA
RNA, Messenger