Mechanism of apoptosis induction in human breast cancer MCF-7 cell by Ruviprase, a small peptide from Daboia russelii russelii venom

Chem Biol Interact. 2016 Oct 25;258:297-304. doi: 10.1016/j.cbi.2016.09.004. Epub 2016 Sep 6.

Abstract

Ruviprase, a 4.4 kDa peptide isolated from Daboia russelii russelii venom demonstrated antiproliferative activity against EMT6/AR1, U-87MG, HeLa and MCF-7 cancer cells with an IC50 value of 23.0, 8.8, 5.8 and 4.0 μg ml(-1), respectively. However, it was nontoxic to non-cancerous human embryonic kidney cell and human peripheral blood lymphocytes. Flow-cytometric analysis confirmed the apoptosis induction in MCF-7 cells by Ruviprase where it induced DNA condensation but did not cause mitotic blockage or chromosomal aberration in treated-cells. Immunofluorescence microscopic analysis indicated Ruviprase induced apoptosis in MCF-7 cells through p53 and p21-mediated pathways. Ruviprase generated reactive oxygen species (ROS), altered the mitochondrial transmembrane potential, and significantly decreased the cellular glutathione (GSH) content of MCF-7 cells. Immunoblotting and quantitative real-time PCR (qRT-PCR) analyses suggested that Ruviprase down-regulated the expression of anti-apoptotic protein Bcl-2, increased cleavage of poly (ADP-ribose) polymerase (PARP) protein, and up-regulated the expression of pro-apoptotic protein Bax, as well as executer protein caspase-7 to induced apoptosis in MCF-7 cells via intrinsic pathway. This is the first report on the characterization of the anticancer potential of a small, non-toxic and anticoagulant peptide purified from Russell's viper venom.

Keywords: Anticancer peptide; Apoptosis; Caspase-7; Mitochondrial pathway; Pro-apoptotic protein; p53.

MeSH terms

  • Apoptosis / drug effects*
  • Blotting, Western
  • Breast Neoplasms / genetics
  • Breast Neoplasms / pathology*
  • Cell Proliferation / drug effects
  • Chromatography, Liquid
  • Female
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Neoplastic / drug effects
  • Glutathione / metabolism
  • Humans
  • Immunoblotting
  • MCF-7 Cells
  • Membrane Potential, Mitochondrial / drug effects
  • Models, Biological
  • Peptides / pharmacology*
  • Reactive Oxygen Species / metabolism
  • Real-Time Polymerase Chain Reaction
  • Snake Venoms / pharmacology*
  • Tandem Mass Spectrometry

Substances

  • Peptides
  • Reactive Oxygen Species
  • Snake Venoms
  • ruviprase, Daboia russelii russelii
  • Glutathione