Induction of Apoptosis by Pierisin-6 in HPV Positive HeLa and HepG2 Cancer Cells is Mediated by the Caspase-3 Dependent Mitochondrial Pathway
(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:
https://lup.lub.lu.se/record/7fc9a569-61d3-42fa-ab63-400878f264df
- author
- Sarathbabu, Subbarayan ; Marimuthu, Satheesh K. ; Ghatak, Souvik LU ; Vidyalakshmi, Subramanian ; Gurusubramanian, Guruswami ; Ghosh, Sankar K. ; Subramanian, Selvi ; Zhang, Wenqing and Senthil Kumar, Nachimuthu
- publishing date
- 2019
- 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}}, }