The fibronectin-derived anti-adhesive peptide III14-2 suppresses adhesion and apoptosis of leukemic cell lines through down-regulation of protein-tyrosine phosphorylation.
We previously found that fibronectin (FN) has a cryptic functional site (YTIYVIAL, #1848-1855) opposing to cell adhesion to extracellular matrix (ECM). The present study demonstrates that the FN peptide containing this anti-adhesive site, termed peptide III14-2, affects programmed cell death (PCD) (apoptosis) as well as cell adhesion by down-regulating protein-tyrosine phosphorylation. Peptide III14-2 suppressed the integrin alpha5beta1-mediated adhesion of leukemic cell lines (K562 and HL60), and protein tyrosine phosphatase inhibitor, 1 microM phenylarsine oxide (PAO) blocked the anti-adhesive effect of peptide III14-2. These leukemic cells underwent PCD when exposed to PAO at the higher concentration (5 microM), as judged by nuclear and DNA fragmentations, and which was reversed by tyrosine kinase inhibitor, genistein. Peptide III14-2 suppressed the PAO-induced PCD, whereas a control peptide in which the anti-adhesive sequence YTIYVIAL is scrambled, was inactive. Western blotting showed that PAO stimulated the tyrosine phosphorylation of cellular proteins including focal adhesion kinase and that peptide III14-2 inhibited them, suggesting that protein-tyrosine phosphorylation represents a common early signal for the adhesion and PCD. The anti-adhesive site of FN molecule may play a crucial role also in a variety of cellular processes other than adhesion and PCD by down-regulating protein-tyrosine phosphorylation.
Fukai
F
F
Department of Patho-Physiology, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Japan. fukai@ps.kagu.sut.ac.jp
Kamiya
S
S
Ohwaki
T
T
Goto
S
S
Akiyama
K
K
Goto
T
T
Katayama
T
T
D017209:Apoptosis / Q000187:drug effects*; D001152:Arsenicals / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects*; D053938:DNA Fragmentation / Q000187:drug effects; D015536:Down-Regulation / Q000187:drug effects; D005353:Fibronectins / Q000737:chemistry*; D019833:Genistein / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D019000:Phosphotyrosine / Q000378:metabolism*; D017469:Receptors, Fibronectin / Q000378:metabolism
D001152:Arsenicals; D005353:Fibronectins; D010446:Peptide Fragments; D017469:Receptors, Fibronectin; C029341:oxophenylarsine; D019000:Phosphotyrosine; D019833:Genistein