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Oxidative stress Attenuates Epithelial Differentiation and Accelerates Cell Migration in Caco‐2 cells
Author(s) -
Dunagan Mitzi,
Rao R.K.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.979.1
Subject(s) - cell migration , occludin , matrigel , microbiology and biotechnology , proto oncogene tyrosine protein kinase src , ly294002 , chemistry , intracellular , cell , tight junction , kinase , biology , biochemistry , phosphatidylinositol
Oxidative stress plays a crucial role in carcinogenesis and tumor metastasis. Numerous studies demonstrated that H 2 O 2 destabilizes intercellular junctions, a process that is involved in cell migration. In the present study, we evaluated the effect of H 2 O 2 on epithelial differentiation and cell migration in Caco‐2 cells. Epithelial differentiation was induced by 3D growth of Caco‐2 cells on matrigel and cell migration analyzed by scratch wound method. H 2 O 2 (100‐250 μM) was administered to the medium. Src kinase inhibitor, PP2 (3 μM) and PI3‐kinase inhibitor, LY294002 (25 μM) were added 30 min prior to H 2 O 2 . The role of c‐Src was determined by stable expression of constitutively active c‐Src(pY418). Cell morphology recorded by phase contrast microscopy and the intercellular junctions assessed by confocal microscopy for occludin and ZO‐1. Results show that H 2 O 2 dose‐dependently accelerated cell migration, which was attenuated by PP2 or LY294002, suggesting that H 2 O 2 promotes cell migration by a c‐Src and PI3‐kinase‐dependent mechanism. H 2 O 2 ‐induced cell migration was associated with redistribution of occludin and ZO‐1 from the intercellular junctions, which was also attenuated by PP2 and LY294002. Expression of c‐Src(pY418) significantly accelerated cell migration. Caco‐2 cells on Matrigel grew to form glandular structure enclosing a cavity. However, in the presence of H 2 O 2 Caco‐2 cells grew into an irregular clump of cells with no cavity; this was associated with irregular distribution of occludin and ZO‐1. These results suggest that oxidative stress may prevent epithelial differentiation and promote cell migration.