Mechanism of apoptosis induction in human breast cancer MCF-7 cell by Ruviprase, a small peptide from Daboia russelii russelii venom.
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.
Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India.
Rupamoni
R
Thakur
Department of Biosciences and Bioengineering, Indian Institute of Technology, Mumbai 400076, Maharashtra, India.
Sudarshan
S
Kini
Molecular Genetics Laboratory, Department of Biotechnology and Bioinformatics, North-Eastern Hill University, Shillong 793022, Meghalaya, India.
Sillarine
S
Kurkalang
Molecular Genetics Laboratory, Department of Biotechnology and Bioinformatics, North-Eastern Hill University, Shillong 793022, Meghalaya, India.
Atanu
A
Banerjee
Molecular Genetics Laboratory, Department of Biotechnology and Bioinformatics, North-Eastern Hill University, Shillong 793022, Meghalaya, India.
Purba
P
Chatterjee
Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India.
Abhishek
A
Chanda
Molecular Genetics Laboratory, Department of Biotechnology and Bioinformatics, North-Eastern Hill University, Shillong 793022, Meghalaya, India.
Anupam
A
Chatterjee
Department of Biosciences and Bioengineering, Indian Institute of Technology, Mumbai 400076, Maharashtra, India.
Dulal
D
Panda
Microbial Biotechnology and Protein Research Laboratory, Department of Molecular Biology and Biotechnology, School of Sciences, Tezpur University, Tezpur 784028, Assam, India. Electronic address: akm@tezu.ernet.in.
Ashis K
AK
Mukherjee
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology*; D049109:Cell Proliferation / Q000187:drug effects; D002853:Chromatography, Liquid; D005260:Female; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D061986:MCF-7 Cells; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008954:Models, Biological; D010455:Peptides / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012910:Snake Venoms / Q000494:pharmacology*; D053719:Tandem Mass Spectrometry
D010455:Peptides; D017382:Reactive Oxygen Species; D012910:Snake Venoms; C000613788:ruviprase, Daboia russelii russelii; D005978:Glutathione
Anticancer peptide; Apoptosis; Caspase-7; Mitochondrial pathway; Pro-apoptotic protein; p53