POTENTIAL ANTITUMOR ACTIVITY OF A BASIC PHOSPHOLIPASE A2 FROM BOTHROPS DIPORUS VENOM

FUSCO, L; MATZNER, V; BUSTILLO S; LEIVA L

Buenos Aires

Jornada; XVIII Jornadas Anuales Multidisciplinarias. Sociedad Argentina de Biología,; 2016

Sociedad Argentina de Biológia

Many toxins from snake venoms have been investigated as possible prototypes for cancer treatment. In particular, different types of phospholipases have been shown to possess antitumor and antiangiogenic properties, such as acidic and basic PLA2s, and synthetic peptides derived from PLA2 homologues. In this work, a basic PLA2 was isolated from Bothrops diporus (yarará chica) venom and its potential antitumor effect in vitro was studied. Purification was made by a two-step procedure utilizing ion exchange (HiTrap SP XL-AKTAprime) and gel filtration chromatography (Sephadex G-75). SDS-PAGE of the isolated enzyme showed a single typical band of 14 kDa and PLA2 activity was evidenced by the formation of hemolytic halos in agarose-erythrocyte egg yolk gels. Cytotoxic activity was determined on a normal (NMuMG) and a tumoral (LM3) epithelial cell lines. Briefly, cells were exposed to different amounts of PLA2 (10-250 g/ml) for 3 h, toxicity was quantitatively assayed by crystal violet method. The percentage of adherent cells in the monolayer culture was registered and its CC50 was calculated. Results indicate that the isolated enzyme PLA2 induces a dose-dependent detachment of cells in both lines, but tumoral LM3 cell line showed to be more sensitivity to the enzyme cytotoxic action (CC50= 13.3 μg/mL) than normal cell line NMuMG (CC50= 74.2 μg/mL). Further studies will be necessaries to demonstrate if this selectivity for tumor cells could be an advantage in the investigation of novel therapeutic agents.