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HO‐2 deletion impairs wound closure in Human Corneal Epithelial (HCE) cells by altering EGFR and FAK‐ mediated signaling pathways
Author(s) -
Halilovic Adna,
Lin Daohong,
Joseph Gregory,
Shkolnik Brian,
Schwartzman Michal L
Publication year - 2013
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.27.1_supplement.765.2
Subject(s) - gene knockdown , transfection , focal adhesion , small hairpin rna , wound healing , microbiology and biotechnology , cell migration , chemistry , cell growth , signal transduction , cell , biology , apoptosis , immunology , biochemistry , gene
Heme oxygenase (HO) represents an intrinsic cytoprotective and anti‐inflammatory system. Inhibition of HO activity significantly impairs wound healing in human corneal epithelial (HCE) cells. HO‐2, but no HO‐1, knockdown using virus‐mediated shRNA plasmid transfection impaired HCE wound healing by 35% and 25% at 24h and 48h after injury. HO‐2 knockdown inhibited HCE migration, measured using a modified Boyden Chamber, by 50% and 40% in control and EGF (10ng/mL) treated media, respectively. The HO‐2 knockdown‐induced inhibition of migration was associated with a 70% decrease in p‐ERK and 35% decrease in p‐Akt levels. Moreover, HO‐2 knockdown reduced p‐EGFR by 45% in injured HCE cells. Immunofluorescence of HO‐2shRNA and control transfected cells revealed a cytoplasmic distribution of p‐FAK. Upon injury, p‐FAK was localized to the nucleus in HO‐2 knockdown cells, whereas, control cells displayed typical p‐FAK as focal adhesion points in the membrane. In summary, HO‐2 knockdown altered EGFR and FAK‐mediated signaling pathways that are crucial for cell migration and proliferation.