Evaluation of cytotoxic and antiproliferative effects and programmed cell death of Venom Disintegrin Jararhagin in human breast adenocarcinoma cells

• Grant number: 12/18256-9
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): April 01, 2013
• Effective date (End): December 31, 2013
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Durvanei Augusto Maria
• Grantee: Manuela Garcia Laveli da Silva
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

The jararhagin is a type of metalloprotease disintegrin isolated from the venom of Bothrops jararaca, comprising metalloproteinase domain, disintegrin (ECD), and rich in cysteine. In vitro studies have shown that treatment with jararhagin decreases cell viability and exhibit inhibitory effects on the adhesion to the substrate of tumor cells, fibroblasts, and endothelial cells. The loss of adhesion occurs by binding the disintegrin-like domain of jararhagin integrins present on the cell membrane, preventing the signaling of extracellular matrix proteins and mediating various cellular phenomena with tumor cells. This design will be assessed in vitro antitumor effects of jararhagin in two cell lines from breast adenocarcinoma MCF-7 and MDA-MB-231 originating from ATCC, with their respective codes HTB-22TM and HTB-26TM. The effects of jararhagin in tumor cell lines will be assessed for cell viability by the MTT colorimetric assay and detection of IC50%, in the formation of free radicals lipoperoxidation, changes in cell death pathways for apoptosis and necrosis, and system for implementing 3D culture (Matrigel). The toxicity of jararhagin will be analyzed by flow cytometry and changes intraestruturais by scanning electron microscopy and confocal laser. Markings lysosomes by fluorescence microscopy using acridine-orange and mitochondrial electric potential probe rhodamine-123 and cytoskeleton arrangement phalloidin by using laser confocal microscopy. The values from the results expressed as mean ± SD and the different statistics will be evaluated by the program Prism version 5.0, being considered values of p <0.05. (AU)