We report the results of a multidisciplinary study on the inhibitory effect of a
snake venom disintegrin,
contortrostatin, a 13.5 kDa homodimeric
protein isolated from Agkistrodon contortrix contortrix (southern copperhead)
venom, on
breast cancer progression. We demonstrate that
contortrostatin binds to
integrins and blocks the adhesion of human
breast cancer cells (MDA-MB-435) to extracellular matrix (ECM)
proteins including
fibronectin and
vitronectin, but it has no effect on adhesion of the cells to
laminin and
Matrigel.
Contortrostatin also prevents invasion of MDA-MB-435 cells through an artificial
Matrigel basement membrane. Daily local injection of
contortrostatin (5 microg per mouse per day) into MDA-MB-435
tumor masses in an orthotopic xenograft nude mouse model inhibits growth of the
tumor by 74% (p = 0.0164). More importantly, it reduces the number of pulmonary macro-
metastasis of the
breast cancer by 68% (p < 0.001), and micro-
metastasis by 62.4% (p < 0.001).
Contortrostatin is not cytotoxic to
cancer cells, and does not inhibit proliferation of the
breast cancer cells in vitro. However,
contortrostatin inhibits angiogenesis induced by the
breast cancer, as shown by immunohistochemical quantitation of the vascular endothelial cells in
tumor tissue removed from the nude mice. We have identified alpha(v)beta3, an important
integrin mediating cell motility and
tumor invasion, as one of the binding sites of
contortrostatin on MDA-MB-435 cells. We conclude that
contortrostatin blocks alpha(v)beta3, and perhaps other
integrins, and thus inhibits in vivo progression.