Interactions of PLA2-s from Vipera lebetina, Vipera berus berus and Naja naja oxiana Venom with Platelets, Bacterial and Cancer Cells

Abstract

Secretory phospholipasesA2 (sPLA2s) form a large family of structurally related enzymes widespread in nature. Herein, we studied the inhibitory effects of sPLA2s from Vipera lebetina (VLPLA2), Vipera berus berus (VBBPLA2), and Naja naja oxiana (NNOPLA2) venoms on (i) human platelets, (ii) four different bacterial strains (gram-negative Escherichia coli and Vibrio fischeri; gram-positive Staphylococcus aureus and Bacillus subtilis) and (iii) five types of cancer cells (PC-3, LNCaP, MCF-7, K-562 and B16-F10) in vitro. sPLA2s inhibited collagen-induced platelet aggregation: VBBPLA2 IC50 = 0.054, VLPLA2 IC50 = 0.072, NNOPLA2 IC50 = 0.814 μM. p-Bromophenacylbromide-inhibited sPLA2 had no inhibitory action on platelets. 36.17 μM VBBPLA2 completely inhibited the growth of gram-positive Bacillus subtilis whereas no growth inhibition was observed towards gram-negative Escherichia coli. The inhibitory action of sPLA2s (~0.7 μM and ~7 μM) towards cancer cells depended on both venom and cell type. VBBPLA2 (7.2 μM) inhibited significantly the viability of K-562 cells and the cell death appeared apoptotic. The sPLA2s exhibited no inhibitory effect towards LNCaP cells and some effect (8%–20%) towards other cells. Thus, already sub-μM concentrations of sPLA2s inhibited collagen-induced platelet aggregation and from the current suite of studied svPLA2s and test cells, VBBPLA2 was the most growth inhibitory towards Bacillus subtilis and K-562 cells.

In Vitro Cytotoxic and Proapoptotic Activities of Anatolian Macrovipera Lebetina Obtusa (Dwigubski, 1832) Crude Venom on Cultured K562 Human Chronic Myelogenous Leukemia Cells.

“…8 Proteins with in vitro and in vivo anticancer activities, including metalloproteinase, disintegrin, PLA 2 , LAAO and CLP have been purified and identified in the venom of different subspecies of M. lebetina. 13,16,19,[36][37][38] These proteins exhibited their cytotoxic effects through different mechanisms such as integrin-blocking and apoptosis induction, resulting in the antiproliferative, antiadhesive and antiangiogenic activities.…”

“…PLA 2 and LAAO from the venom of Vipera berus berus also showed proapoptotic effect on K562 cells. 38,42 Nunes et al 17 reported apoptosis inducing effect of CLP from Bothrops lecurus venom on K562 cells. Furthermore, venoms or purified proteins from different subspecies of M. lebetina also induced apoptosis in other various cancers and non-cancerous cells, through different mechanisms.…”

“…12,42 PLA 2 from the venoms of M. lebetina turanica, Vipera berus berus and Naja naja oxiana have shown to inhibit the viability of K562 cells selectively among other cancer cell lines (LnCaP, PC-3, MCF-7, B10-F16); however, PLA 2 from M. lebetina turanica had only a slight effect compared to the other two species. 38 Conceição et al 43 reported that jararhagin, a P-III metalloproteinase from the venom of Bothrops jararaca decreased the viability of K562 cells via inducing autophagy and apoptosis. Debnath et al…”

“…32,33 There are limited studies investigating the induction of apoptosis in K562 cells, after treatment with crude snake venom or purified proteins. 14,17,19,22,38,[41][42][43] Samel et al 19 investigated the effect of Sephadex G-100 chromatography fractions of M. lebetina turanica (subspecies was not mentioned in the paper but assigned according to the locality) venom and also purified HUVEC apoptosis inducing metalloproteinase (VLAIP). The first fraction reported to inhibit the viability of K562 cells about 30%.…”

Antimicrobial activity of apitoxin, melittin and phospholipase A2 of honey bee (Apis mellifera) venom against oral pathogens

“…According to Samel et al (2013), phospholipases A 2 are enzymes that catalyze the hydrolysis of the sn-2 fatty acyl ester bond of sn-3 phosphoglycerides, liberating free fatty acids, and lysophospholipids. Phospholipases A 2 (PLA 2 s) are a large family of proteins found in various mammalian tissues, arthropods, and in the venoms of snakes, scorpions, and bees.…”