Regulation of tumor cell chemotaxis by type IV collagen is mediated by a Ca2+-dependent mechanism requiring CD47 and the integrin alpha(v)beta(3)

Studies from our laboratories demonstrated that synthetic peptides from the non-collagenous (NC-1) domain of the (alpha 3 (IV) chain of type TV collag en (COL IV) enhanced tumor cell adhesion (Han, J., Ohno, N., Monboisse, J. C., Pasco, S., Borel, J. P., and Kefalides, N. A. (1997) J. Biol. Chem. 272 , 20395-20401). We have isolated the receptors for the alpha 3(IV)185-203 p eptide from melanoma and prostate tumor cells and identified them as CD47/i ntegrin-associated protein and the integrin alpha(v)beta(3) (Shahan, T. A, Ziaie, Z., Pasco, S., Fawzi, A, Bellon, G., Monboisse, J. C., and Kefalides , N. A. (1999) Cancer Res. 59, 4584-4590). In the present study we have exa mined the effect of CD47 and the integrin alpha(v)beta(3) on in vitro tumor cell chemotaxis and Ca-i(2+) modulation in response to COL IV, from the an terior lens capsule (ALC-COL IV) and peptides from its NC-1 domain. COL TV as well as the alpha 3(IV) peptide promoted tumor cell chemotaxis with an i mmediate increase in intracellular [Ca2+]. Treating tumor cells with CD47 a nd integrin alpha(v)beta(3)-reactive antibodies reduced chemotaxis as well as the rise in [Ca2+](i) in response to ALC-COL IV or the alpha 3(IV)185-20 3 peptide but not to Engelbreth-Holm-Swarm-COL IV or fibronectin. The a3(IV )185-203 synthetic peptide stimulated an increase in calcium from intracell ular stores exclusively, whereas ALC-COL IV, Engelbreth-Holm-Swarm-COL IV, and fibronectin stimulated Ca2+ flux from both internal and external stores . Furthermore, treatment of the cells with Ca2+ chelator bis-(O-aminophenox yl)ethane-N,N,N',N' -tetraaceticacid-acetomethoxy ester inhibited chemotaxi s toward both ALC-COL IV and the alpha 3(IV)185-203 peptide. These data ind icate that CD47 and integrin alpha(v)beta(3) regulate tumor cell chemotaxis in response to COL IV and the alpha 3(IV)185-203 peptide through a Ca2+-de pendent mechanism.