Mechanism of apoptosis induction in human breast cancer MCF-7 cell by Ruviprase, a small peptide from Daboia russelii russelii venom. (25th October 2016)

• Record Type: Journal Article
• Authors: Thakur, Rupamoni; Kini, Sudarshan; Kurkalang, Sillarine; Banerjee, Atanu; Chatterjee, Purba; Chanda, Abhishek; Chatterjee, Anupam; Panda, Dulal; Mukherjee, Ashis K.
• Is Part Of: Chemico-biological interactions. Volume 258(2016)
• Page Start: 297
• Page End: 304
• Publication Date: 2016-10-25
• Subjects: Anticancer peptide -- Apoptosis -- Caspase-7 -- Mitochondrial pathway -- p53 -- Pro-apoptotic protein; BSA bovine serum albumin -- DIC differential interference contrast -- EMEM Eagle's minimal essential medium -- GSH cellular glutathione -- HPBL human peripheral blood lymphocytes -- PARP poly (ADP-ribose) polymerase -- PBS phosphate buffered saline -- PI propidium iodide -- qRT-PCR Quantitative Real-time PCR -- ROS reactive oxygen species -- RVV Russell's Viper venom; Biochemistry -- Periodicals; Toxicological chemistry -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biochimie -- Périodiques; Toxicologie biochimique -- Périodiques; 572
• DOI: 10.1016/j.cbi.2016.09.004
• Languages: English
• ISSNs: 0009-2797

Abstract

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. Highlights: Ruviprase shows selective cytotoxicity towards MCF-7 cells. Ruviprase leads to p53-mediated cell-cycle arrest in MCF-7 cells. Ruviprase generates ROS and alters mitochondrial transmembrane potential. Ruviprase up-regulates Bax, caspase 7 expression besides increasing PARP cleavage. Ruviprase is a promising molecule for development of peptide-based anticancer drug.