Snake venom phospholipase A2 (svPLA2) from Crotalus durissus terrificus inhibits tumoral cell adhesion

FACUNDO SPIAZZI; LAURA D. GALARZA; DANIELA SASOVSKY; GUILLERMO ESCOBAR ; LUCIANO FUSCO; LAURA C. LEIVA; SILVINA ETCHEVERRIA; SOLEDAD BUSTILLO

Mar del Plata

Congreso; Reunión Anual de SOCIEDADES DE BIOCIENCIA SAIC.SAFE.SAB.SAP. AACYTAL. NANOMED-ar. HCS; 2019

S0ciedades de Biociencia

Snake venoms are natural sources of bioactive substances with therapeutic potential.In particular, different types of phospholipases have been shown to possess antitumorand antiangiogenic properties. The venom of the South American Rattlesnake,Crotalus durissus terrificus (C.d.t.), is a complex mixture of proteins. Crotoxin, the maintoxin of this venom, contains a basic phospholipase A2 (PLA2) and a non-toxic acidicprotein, crotapotin. Thus, the aim of this study was to evaluate the potential effect of aPLA2 isolated from C.d.t. (Cdt-PLA2) on tumoral cell adhesion.Cdt-PLA2 was purified by two chromatographic steps, a gel filtration and a reversedphase HPLC C-18 chromatographs. The purity of the enzyme was verified by SDSPAGE.Firstly, cytotoxicity of Cdt-PLA2 (5 - 250 μg/mL) was assessed in cultured murinetumoral epithelial cell line, LM3, grown in DMEM-5% FBS at 37°C-5% CO2. Noncytotoxic concentrations were selected for adhesion inhibition assay. Briefly, LM3 cells(3×104/well) were preincubated for 30 min at 37°C with Cdt-PLA2 (5, 10, 20, 25, and 50μg/mL) or culture medium (control) and then added to 96-well plates. After 1.5 h, nonadherent cells were removed by careful washing and aspiration with PBS. Adherentcells were fixed and stained with crystal violet. The percentage of cell adhesion wasdetermined by comparison of the absorbance readings (620nm) with the meanabsorbance of control cells (not exposed to the PLA2s), considered as 100% adhesion.The obtained results showed a concentration-dependent adhesion inhibition effect ofCdt-PLA2 using non-cytotoxic concentrations. These findings are the beginning of thestudy of the potential use of this enzyme as an antitumoral.