Pro-apoptotic signaling pathway activated by echistatin in GD25 cells.
Disintegrins, low molecular weight RGD-containing polypeptides isolated from snake venoms, have seen use as integrin antagonists in the field of tumor biology and angiogenesis. In this study, we investigated the molecular mechanism by which the disintegrin echistatin affects cell adhesion and signaling resulting in an apoptotic response in the GD25 cell system. Wild-type GD25 cells, which lack expression of the beta(1) family of integrin, and stable transfectants expressing the A isoform of beta(1) integrin subunit were used. Nanomolar concentrations of echistatin detached fibronectin- and vitronectin-adherent GD25 cells from immobilized substratum. However, prior to inducing detachment of adherent cells, echistatin caused apoptosis as measured by caspase-3 activation. Either cell detachment or apoptotic response induced by echistatin were more pronounced on fibronectin-adherent GD25 cells than on vitronectin-adherent ones. GD25 cell exposure to echistatin caused a reduction of tyrosine phosphorylation levels of pp125(FAK), whereas it didn't affect either Shc tyrosine phosphorylation levels or Shc-Grb2 functional association. The down-regulation of pp125(FAK) preceded apoptosis and cell detachment induced by echistatin. Our results indicate that pp125(FAK) and not Shc pathway is involved in echistatin-induced apoptotic response in the GD25 cell system.
Dipartimento di Strutture, Funzioni e Tecnologie Biologiche, Università di Napoli Federico II, Naples, Italy.
Elena
E
Alimenti
Simona
S
Tafuri
Antonio
A
Scibelli
Danila
D
D'Angelo
Laura
L
Manna
Luigi Michele
LM
Pavone
M Antonietta
MA
Belisario
Norma
N
Staiano
D017209:Apoptosis / Q000187:drug effects*; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D005353:Fibronectins / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D019096:Vitronectin / Q000378:metabolism
D005353:Fibronectins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D019096:Vitronectin; C057986:echistatin