Functional Effect of Contortrostatin, a Snake Venom Disintegrin, on Human Glioma Cell Invasion In Vitro

Abstract

The metastatic spread of cancer is a complex process that involves the combination of different cellular actions including cell adhesion to the extracellular matrix (ECM), breakdown of the ECM by specific matrix-degrading proteinases, and active cell locomotion. Contortrostatin (CN), a homodimeric snake venom disintegrin, has previously been demonstrated to be effective in blocking vitronectin/fibronectin-dependent adhesion and invasion of T98G human glioblastoma cells through Matrigel using in vitro studies. However, it is not known at what step of the invasion process CN exerts its inhibitory effect. In the present report, CN is shown to decrease invasion of various glioma cell lines through Matrigel affecting neither cell adhesion, nor cell viability. While CN had no effect on cell binding to laminin and type IV collagen, it blocked adhesion of alphav beta3-positive, but not alphav beta3-negative cells, to vitronectin and fibronectin. Furthermore, members of the matrix metalloproteinase (MMP) family and their physiological inhibitors, and of the plasminogen activator (PA)/plasmin system were demonstrated not to be involved in CN-induced loss of glioma cell invasiveness. Instead, CN inhibited active locomotion of cells on Matrigel. These data suggest that CN-mediated inhibition of glioma cell invasion through Matrigel is a direct result of impaired cell motility. Moreover, use of several glioma cell lines and integrin antibodies strongly indicates the versatility of CN in inhibiting the invasion process based on the ability of CN to interact with different integrins, including alphav beta3, alphav beta5, and alpha5beta1.

Action of the Disintegrin Contortrostatin on Breast Cancer Cell Primary Cultures

“…Studies employing cell lines have shown that contortrostatin can be effectively used in vitro in a number of cases [37], but there is no information about the effect of this drug in a primary culture of breast cancer cell where tumor cells retain their original phenotype. The results in our study are in agreement with data provided by other workers [27,35] and they extend our previous observations [25,38] on the use of RGD peptides in primary breast cancer cultures.…”

“…Snake venoms and hemostatic system have been under consideration in relation to the role of disintegrins in platelet aggregation [26]. The anti-invasive activity of contortrostatin has been studied using human glioblastoma cell lines [27] as well as its anti-tumor activity in Ehrich ascites carcinoma [28].…”

Evaluating kinetic and physiological features of rCHO-K1 cells cultured on microcarriers for production of a recombinant metalloprotease/disintegrin

“…Snake venom disintegrins are synthesized in vivo as multimodular proteins that comprise a signal peptide, a pro-domain, a metalloprotease domain, a disintegrin domain, and a cysteine-rich domain (Bjarnason and Fox, 1994;Fox and Serrano, 2005;Ramos and Selistre-de-Araujo, 2006). Recently, several disintegrins have been proposed as therapeutic drugs, in view of their antitumor, antiangiogenic and antithrombotic activities (Braud et al 2000;Matsui et al 2000;Zhou et al 2000;Schmitmeier et al 2003;Swenson et al 2004;Markland et al 2005;Yang et al 2005). For such applications, a few cDNAs encoding disintegrins, originally from snake venom glands, have been cloned and expressed both in bacteria and in insect cells (Park et al 1998;Fan et al 1999;Jeon and Kim, 1999;Wang et al 2003;Wang et al 2004) by genetic engineering techniques.…”

Serum from rabbit orally administered cobra venom inhibits growth of implanted hepatocellular carcinoma cells in mice

“…Its toxic and side effects were unavoidable. Great efforts have been made [42][43][44][45] , but the problem remains unresolved. So a long-standing goal in snake venom therapy of cancer is to find a stable, low toxic, highly effective chemotherapeutic agent that selectively targets tumor cells.…”