Cell migration inhibition activity of a non-RGD disintegrin from Crotalus durissus collilineatus venom

Abstract

BackgroundIn recent decades, snake venom disintegrins have received special attention due to their potential use in anticancer therapy. Disintegrins are small and cysteine-rich proteins present in snake venoms and can interact with specific integrins to inhibit their activities in cell-cell and cell-ECM interactions. These molecules, known to inhibit platelet aggregation, are also capable of interacting with certain cancer-related integrins, and may interfere in important processes involved in carcinogenesis. Therefore, disintegrin from Crotalus durissus collilineatus venom was isolated, structurally characterized and evaluated for its toxicity and ability to interfere with cell proliferation and migration in MDA-MB-231, a human breast cancer cell line.MethodsBased on previous studies, disintegrin was isolated by FPLC, through two chromatographic steps, both on reversed phase C-18 columns. The isolated disintegrin was structurally characterized by Tris-Tricine-SDS-PAGE, mass spectrometry and N-terminal sequencing. For the functional assays, MTT and wound-healing assays were performed in order to investigate cytotoxicity and effect on cell migration in vitro, respectively.ResultsDisintegrin presented a molecular mass of 7287.4 Da and its amino acid sequence shared similarity with the disintegrin domain of P-II metalloproteases. Using functional assays, the disintegrin showed low cytotoxicity (15% and 17%, at 3 and 6 μg/mL, respectively) after 24 h of incubation and in the wound-healing assay, the disintegrin (3 μg/mL) was able to significantly inhibit cell migration (24%, p < 0.05), compared to negative control.ConclusionThus, our results demonstrate that non-RGD disintegrin from C. d. collilineatus induces low cytotoxicity and inhibits migration of human breast cancer cells. Therefore, it may be a very useful molecular tool for understanding ECM-cell interaction cancer-related mechanisms involved in an important integrin family that highlights molecular aspects of tumorigenesis. Also, non-RGD disintegrin has potential to serve as an agent in anticancer therapy or adjuvant component combined with other anticancer drugs.

Anticancer potential of nanogold conjugated toxin GNP-NN-32 from Naja naja venom

“…In this context, this study indicated that GNP-NN 32 was capable of inducing apoptosis at a concentration of 1.5 μg/mL in MCF-7 cell line and 5 μg/mL in MDA-MB-231 cell line. Our results are in agreement with other studies that discovered that disintegrin from Crotalus durissus collilineatus venom induced cytotoxicity [37] whereas BthTX-I from Bothrops jararacussu induced apoptosis [38] and also showed antitumor and antimetastatic effects [39] on human breast cancer cell lines.…”

Cytotoxic and anticancer properties of the Malaysian mangrove pit viper (Trimeresurus purpureomaculatus) venom and its disintegrin (purpureomaculin)

“…The antineoplastic potential of snake venom disintegrins have been shown in several studies, e.g. contortrostatin (disintegrin isolated from Agkistrodon contortrix venom) [23], saxatilin (disintegrin from Gloydius saxatilis venom) [24], PAIEM (disintegrin isolated from Echis multisquamatis venom) [25] and a disintegrin from Crotalus durissus collilineatus venom [26]. The disintegrins act by inhibiting angiogenesis, invasion and migration of cancer cells.…”

The Effects of αvβ3 Integrin Blockage in Breast Tumor and Endothelial Cells under Hypoxia In Vitro

“…These studies, however, were performed in normoxia only. Non-RGD disintegrins from Crotalus durissus colineatus inhibited MDA-MB-231 migration in a wound healing assay after 24 h in normoxia [61].…”