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7116090 
Journal Article 
CAP37 Activation of PKC Promotes Human Corneal Epithelial Cell Chemotaxis 
Griffith, GL; Russell, RA; Kasus-Jacobi, A; Thavathiru, E; Gonzalez, ML; Logan, S; Pereira, HA; , 
2013 
Yes 
Investigative Ophthalmology and Visual Science
ISSN: 0146-0404
EISSN: 1552-5783 
ASSOC RESEARCH VISION OPHTHALMOLOGY INC 
ROCKVILLE 
54 
10 
6712-6723 
English 
24008408 
WOS:000326567700030 
PURPOSE. The objective of this study was to elucidate the signaling pathway through which cationic antimicrobial protein of 37 kDa (CAP37) mediates human corneal epithelial cell (HCEC) chemotaxis.METHODS. Immortalized HCECs were treated with pertussis toxin (10 and 1000 ng/mL), protein kinase C (PKC) inhibitors (calphostin c, 50 nM and Ro-31-8220, 100 nM), phorbol esters (phorbol 12,13-dibutyrate, 200 nM and phorbol 12-myristate 13-acetate, 1 mu M) known to deplete PKC isoforms, and siRNAs (400 nM) before a modified Boyden chamber assay was used to determine the effect of these inhibitors and siRNAs on CAP37-directed HCEC migration. PKC delta protein levels, PKC delta-Thr(505) phosphorylation, and PKC delta kinase activity was assessed in CAP37-treated HCECs using immunohistochemistry, Western blotting, and a kinase activity assay, respectively.RESULTS. Chemotaxis studies revealed that treatment with pertussis toxin, PKC inhibitors, phorbol esters, and siRNAs significantly inhibited CAP37-mediated chemotaxis compared with untreated controls. CAP37 treatment increased PKC delta protein levels and led to PKC delta phosphorylation on residue Thr(505). Direct activation of PKC delta by CAP37 was demonstrated using a kinase activity assay.CONCLUSIONS. These findings lead us to conclude that CAP37 is an important regulator of corneal epithelial cell migration and mediates its effects through PKC delta.