Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Induction of Apoptosis by Pierisin-6 in HPV Positive HeLa and HepG2 Cancer Cells is Mediated by the Caspase-3 Dependent Mitochondrial Pathway

Sarathbabu, Subbarayan ; Marimuthu, Satheesh K. ; Ghatak, Souvik LU ; Vidyalakshmi, Subramanian ; Gurusubramanian, Guruswami ; Ghosh, Sankar K. ; Subramanian, Selvi ; Zhang, Wenqing and Senthil Kumar, Nachimuthu (2019) In Anti-Cancer Agents in Medicinal Chemistry 19(3). p.337-346
Abstract
Background
To explore the cytotoxic and apoptotic activity of the pierisin-6 protein in HPV HeLa and HepG2 cell lines.
Methods
In this study, isolation, and purification of cytotoxic Prierisin-6 from the larvae of Pieris napi by affinity column chromatography techniques. Characterization of full-length mRNA of pierisin-6 gene was performed using 3'/5' RACE PCR. The quantitative RT-PCR used to study the developmental stage-specific expression of pierisin-6 mRNA. The most effective concentration of Pierisin-6 protein was determined by measuring cell proliferation. Apoptosis was assessed using AO/Et-Br, Propidium Iodide, and Rhodamine 123 assays, whereas protein levels of caspase 3, cytochrome C were evaluated by ELISA method.... (More)
Background
To explore the cytotoxic and apoptotic activity of the pierisin-6 protein in HPV HeLa and HepG2 cell lines.
Methods
In this study, isolation, and purification of cytotoxic Prierisin-6 from the larvae of Pieris napi by affinity column chromatography techniques. Characterization of full-length mRNA of pierisin-6 gene was performed using 3'/5' RACE PCR. The quantitative RT-PCR used to study the developmental stage-specific expression of pierisin-6 mRNA. The most effective concentration of Pierisin-6 protein was determined by measuring cell proliferation. Apoptosis was assessed using AO/Et-Br, Propidium Iodide, and Rhodamine 123 assays, whereas protein levels of caspase 3, cytochrome C were evaluated by ELISA method. Pierisin-6 induced cell cycle arrest was determined using Propidium iodide by FACS.
Results
In this study, Pierisin-6, a novel apoptotic protein was found to have cytotoxicity against HeLa, HepG2 human cancer cell lines and L-132 human lung epithelial cell line. Among the target cells, HeLa was the most sensitive to Pierisin-6. Flow cytometry analysis confirms an increased percentage of apoptotic cells in sub G1 phase and cell cycle arrest at S phase. Alteration in the transmembrane potential of mitochondria, Cytochrome c released from the mitochondrial membrane, and caspase substrate assay demonstrated the cleavage of Ac- DEVD-pNA signifying the activation of Caspase-3. These findings suggested that Pierisin-6 significantly induce apoptosis in HeLa and HepG2 cells and is attributed mainly through a mitochondrial pathway by activation of caspases. The developmental and stage-specific expression of pierisin-6 mRNA was one thousand-fold increased from second to third instar larvae and gradually declined before pupation.
Conclusion
Pierisin-6 represents a promising therapeutic approach for liver cancer patients. (Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; and
publishing date
type
Contribution to journal
publication status
published
in
Anti-Cancer Agents in Medicinal Chemistry
volume
19
issue
3
pages
337 - 346
publisher
Bentham Science Publishers
external identifiers
  • scopus:85069458530
ISSN
1871-5206
DOI
10.2174/1871520619666181127113848
language
English
LU publication?
no
id
7fc9a569-61d3-42fa-ab63-400878f264df
date added to LUP
2021-11-09 15:56:37
date last changed
2022-04-19 17:50:03
@article{7fc9a569-61d3-42fa-ab63-400878f264df,
  abstract     = {{Background<br/>To explore the cytotoxic and apoptotic activity of the pierisin-6 protein in HPV HeLa and HepG2 cell lines.<br/>Methods<br/>In this study, isolation, and purification of cytotoxic Prierisin-6 from the larvae of Pieris napi by affinity column chromatography techniques. Characterization of full-length mRNA of pierisin-6 gene was performed using 3'/5' RACE PCR. The quantitative RT-PCR used to study the developmental stage-specific expression of pierisin-6 mRNA. The most effective concentration of Pierisin-6 protein was determined by measuring cell proliferation. Apoptosis was assessed using AO/Et-Br, Propidium Iodide, and Rhodamine 123 assays, whereas protein levels of caspase 3, cytochrome C were evaluated by ELISA method. Pierisin-6 induced cell cycle arrest was determined using Propidium iodide by FACS.<br/>Results<br/>In this study, Pierisin-6, a novel apoptotic protein was found to have cytotoxicity against HeLa, HepG2 human cancer cell lines and L-132 human lung epithelial cell line. Among the target cells, HeLa was the most sensitive to Pierisin-6. Flow cytometry analysis confirms an increased percentage of apoptotic cells in sub G1 phase and cell cycle arrest at S phase. Alteration in the transmembrane potential of mitochondria, Cytochrome c released from the mitochondrial membrane, and caspase substrate assay demonstrated the cleavage of Ac- DEVD-pNA signifying the activation of Caspase-3. These findings suggested that Pierisin-6 significantly induce apoptosis in HeLa and HepG2 cells and is attributed mainly through a mitochondrial pathway by activation of caspases. The developmental and stage-specific expression of pierisin-6 mRNA was one thousand-fold increased from second to third instar larvae and gradually declined before pupation.<br/>Conclusion<br/>Pierisin-6 represents a promising therapeutic approach for liver cancer patients.}},
  author       = {{Sarathbabu, Subbarayan and Marimuthu, Satheesh K. and Ghatak, Souvik and Vidyalakshmi, Subramanian and Gurusubramanian, Guruswami and Ghosh, Sankar K. and Subramanian, Selvi and Zhang, Wenqing and Senthil Kumar, Nachimuthu}},
  issn         = {{1871-5206}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{337--346}},
  publisher    = {{Bentham Science Publishers}},
  series       = {{Anti-Cancer Agents in Medicinal Chemistry}},
  title        = {{Induction of Apoptosis by Pierisin-6 in HPV Positive HeLa and HepG2 Cancer Cells is Mediated by the Caspase-3 Dependent Mitochondrial Pathway}},
  url          = {{http://dx.doi.org/10.2174/1871520619666181127113848}},
  doi          = {{10.2174/1871520619666181127113848}},
  volume       = {{19}},
  year         = {{2019}},
}