Anti-invasive and anti-adhesive activities of a recombinant disintegrin, r-viridistatin 2, derived from the Prairie rattlesnake (Crotalus viridis viridis)

• Published in: Toxicon (Oxford) 2012-07, Vol.60 (1), p.31-39
• Main Authors: Lucena, Sara E., Jia, Ying, Soto, Julio G., Parral, Jessica, Cantu, Esteban, Brannon, Jeremy, Lardner, Kristina, Ramos, Carla J., Seoane, Agustin I., Sánchez, Elda E.
• Format: Article
• Language: English
• Subjects: adenocarcinoma; adhesion; Amino Acid Sequence; amino acids; Animal poisons toxicology. Antivenoms; Animals; Anti-metastatic activity; anticarcinogenic activity; antineoplastic agents; Base Sequence; basement membrane; Biological and medical sciences; bladder; Cancer cell lines; Cell Adhesion - drug effects; Cell Line, Tumor; Crotalid Venoms - chemistry; Crotalus; Crotalus viridis; Crotalus viridis viridis; Disintegrin; Disintegrins - chemistry; Disintegrins - pharmacology; DNA Primers; DNA, Complementary; extracellular matrix; fibrosarcoma; Humans; Medical sciences; melanoma; Mice; Molecular Sequence Data; Neoplasm Invasiveness; platelet aggregation; Platelet Aggregation Inhibitors - pharmacology; proteins; Recombinant Proteins - chemistry; Recombinant Proteins - pharmacology; Recombinant snake toxins; Sequence Homology, Amino Acid; Toxicology; venoms
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/j.toxicon.2012.03.011

Snake venom disintegrins inhibit platelet aggregation and have anti-cancer activities. In this study, we report the cloning, expression, and functional activities of a recombinant disintegrin, r-viridistatin 2 (GenBank ID: JQ071899), from the Prairie rattlesnake. r-Viridistatin 2 was tested for anti-invasive and anti-adhesive activities against six different cancer cell lines (human urinary bladder carcinoma (T24), human fibrosarcoma (HT-1080), human skin melanoma (SK-Mel-28), human colorectal adenocarcinoma (CaCo-2), human breast adenocarcinoma (MDA-MB-231) and murine skin melanoma (B16F10)). r-Viridistatin 2 shares 96% and 64% amino acid identity with two other Prairie rattlesnake medium-sized disintegrins, viridin and viridistatin, respectively. r-Viridistatin 2 was able to inhibit adhesion of T24, SK-MEL-28, HT-1080, CaCo-2 and MDA-MB-231 to various extracellular matrix proteins with different affinities. r-Viridistatin 2 decreased the ability of T24 and SK-MEL-28 cells to migrate by 62 and 96% respectively, after 24 h of incubation and the invasion of T24, SK-MEL-28, HT-1080 and MDA-MB-231 cells were inhibited by 80, 85, 65 and 64% respectively, through a reconstituted basement membrane using a modified Boyden chamber. Finally, r-viridistatin 2 effectively inhibited lung colonization of murine melanoma cells in BALB/c mice by 71%, suggesting that r-viridistatin 2 could be a potent anti-cancer agent in vivo. ► A recombinant disintegrin, r-viridistatin 2, from the Prairie rattlesnake was cloned, expressed and isolated. ► r-Viridistatin 2 was able to inhibit the adhesion of 5 different cancer cell lines to various extracellular matrices. ► r-Viridistatin 2 effectively inhibited tumor colonization in mice by 71%.