Human breast cancer cell death induced by BnSP-6, a Lys-49 PLA₂ homologue from Bothrops pauloensis venom

Fernanda Van Petten Vasconcelos Azevedo; Daiana Silva Lopes; Sarah Natalie Cirilo Gimenes; David Collares Achê; Lara Vecchi; Patrícia Terra Alves; Denise de Oliveira Guimarães; Renata Santos Rodrigues; Luiz Ricardo Goulart; Veridiana de Melo Rodrigues; Kelly Aparecida Geraldo Yoneyama

doi:10.1016/j.ijbiomac.2015.10.080

Human breast cancer cell death induced by BnSP-6, a Lys-49 PLA₂ homologue from Bothrops pauloensis venom

Int J Biol Macromol. 2016 Jan:82:671-7. doi: 10.1016/j.ijbiomac.2015.10.080. Epub 2015 Oct 28.

Affiliations

¹ Laboratory of Biochemistry and Animal Toxins, Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil.
² Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil.
³ Laboratory of Biochemistry and Animal Toxins, Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil; INCT, Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica, Belo Horizonte, Brazil.
⁴ Laboratory of Biochemistry and Animal Toxins, Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil; INCT, Instituto Nacional de Ciência e Tecnologia em Nano-Biofarmacêutica, Belo Horizonte, Brazil. Electronic address: veridiana@ingeb.ufu.br.
⁵ Laboratory of Biochemistry and Animal Toxins, Institute of Genetics and Biochemistry, Federal University of Uberlandia, UFU, Uberlandia, MG, Brazil. Electronic address: kelly@ingeb.ufu.br.

PMID: 26519876
DOI: 10.1016/j.ijbiomac.2015.10.080

Abstract

This work shows the antitumoral effects of BnSP-6, a Lys 49 PLA2 isolated from Bothrops pauloensis venom, on human breast cancer MDA-MB-231 cells. BnSP-6 caused a dose-dependent cytotoxicity and inhibited cell adhesion. Interestingly, cytotoxic activity of BnSP-6 was significantly lower against MCF10A, a non-tumorigenic breast cell line, suggesting that this PLA2 presented a possible preference for targets in cancer cells. Analysis of cell death on MDA-MB-231 cells showed that BnSP-6 stimulated the autophagy process, as evidenced by labeling of autophagic vacuoles. Moreover, apoptosis assays showed that BnSP-6 induced both early and late apoptosis. Apoptosis of MDA-MB-231 cells was also confirmed by up-regulation of different genes related to the apoptosis pathway, such as TNF, TNFRSF10B, TNFRSF1A and CASP8 and decreased expression of anti-apoptotic genes (BCL2 and BCL2L). In addition, BnSP-6 caused a remarkable increase in gene expression of BRCA2 and TP53 tumor suppressors. Finally, BnSP-6 induced down-regulation of Angiopoetin 1 gene (potent pro-angiogenic factor) and inhibited adhesion and migration of MDA-MB-231 cells suggesting pharmaceutical applications of this PLA2 as an antiangiogenic and anti-metastatic agent. Taken together, our results show that the PLA2 BnSP-6 presents anticancer potential that can be exploited as prototype for the design of new therapies.

Keywords: Apoptosis; Autophagy; Breast cancer; Cell migration; Phospholipase A(2); Snake venom.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects
Autophagy / drug effects
Bothrops / metabolism*
Breast Neoplasms
Cell Adhesion / drug effects
Cell Death / drug effects
Cell Line, Tumor
Cell Movement
Crotalid Venoms / chemistry*
Crotalid Venoms / isolation & purification
Crotalid Venoms / toxicity*
Female
Gene Expression
Humans
Phospholipases A2 / chemistry*
Phospholipases A2 / isolation & purification
Phospholipases A2 / toxicity*
Protein Interaction Domains and Motifs

Substances

Crotalid Venoms
BnSP-6
Phospholipases A2