Cargando…
Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade
Death Receptor 5 (DR5) is known to be an important anti-cancer drug target. TRAIL is a natural ligand of DR5, but its drug action is limited because of several factors. A few agonistic ligands were identified as TRAIL-DR5 axis modulators, which enhance the cellular apoptosis. Literature suggest that...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352116/ https://www.ncbi.nlm.nih.gov/pubmed/27542249 http://dx.doi.org/10.18632/oncotarget.11335 |
_version_ | 1782514887585431552 |
---|---|
author | Das, Sarita Tripathi, Neha Preet, Ranjan Siddharth, Sumit Nayak, Anmada Bharatam, Prasad V. Kundu, Chanakya Nath |
author_facet | Das, Sarita Tripathi, Neha Preet, Ranjan Siddharth, Sumit Nayak, Anmada Bharatam, Prasad V. Kundu, Chanakya Nath |
author_sort | Das, Sarita |
collection | PubMed |
description | Death Receptor 5 (DR5) is known to be an important anti-cancer drug target. TRAIL is a natural ligand of DR5, but its drug action is limited because of several factors. A few agonistic ligands were identified as TRAIL-DR5 axis modulators, which enhance the cellular apoptosis. Literature suggest that quinacrine (QC) acts as a DR5 agonistic ligand. However, the detailed mechanism explaining how QC interacts with TRAIL-DR5 axis has not been established. Also focused in vitro and in vivo experimental analysis to validate the hypothesis is not yet performed. In this work, extensive studies have been carried out using in silico analysis (molecular dynamics), in vitro analysis (cell based assays) and in vivo analysis (based on mice xenograft model), to delineate the mechanism of QC action in modulating the TRAIL-DR5 signaling. The MD simulations helped in identifying the important residues contributing to the formation of a QC-TRAIL-DR5 complex, which provide extra stability to it, consequently leading to the enhanced cellular apoptosis. QC caused a dose dependent increase of DR5 expression in cancer cells but not in normal breast epithelial cells, MCF-10A. QC showed a synergistic effect with TRAIL in causing cancer cell apoptosis. In DR5-KD MCF-10A-Tr (DR5 knocked down) cells, TRAIL+ QC failed to significantly increase the apoptosis but over expression of full length DR5 in DR5-silence cells induced apoptosis, further supporting DR5 as a drug target for QC. An increase in the release of reactive species (ROS and RNS) and activation of enzymes (FADD, CASPASES 3, 8, 9 and cytochrome-C) indicated the involvement of mitochondrial intrinsic pathway in TRAIL+QC mediated apoptosis. In vivo study pointed out that TRAIL+QC co-administration increases the expression of DR5 and reduce the tumor size in xenograft mice. This combined in silico, in vitro and in vivo analysis revealed that QC enhances the cellular apoptosis via the modulation of TRAIL-DR5 complexation and the mitochondrial intrinsic pathway. |
format | Online Article Text |
id | pubmed-5352116 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-53521162017-04-13 Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade Das, Sarita Tripathi, Neha Preet, Ranjan Siddharth, Sumit Nayak, Anmada Bharatam, Prasad V. Kundu, Chanakya Nath Oncotarget Research Paper Death Receptor 5 (DR5) is known to be an important anti-cancer drug target. TRAIL is a natural ligand of DR5, but its drug action is limited because of several factors. A few agonistic ligands were identified as TRAIL-DR5 axis modulators, which enhance the cellular apoptosis. Literature suggest that quinacrine (QC) acts as a DR5 agonistic ligand. However, the detailed mechanism explaining how QC interacts with TRAIL-DR5 axis has not been established. Also focused in vitro and in vivo experimental analysis to validate the hypothesis is not yet performed. In this work, extensive studies have been carried out using in silico analysis (molecular dynamics), in vitro analysis (cell based assays) and in vivo analysis (based on mice xenograft model), to delineate the mechanism of QC action in modulating the TRAIL-DR5 signaling. The MD simulations helped in identifying the important residues contributing to the formation of a QC-TRAIL-DR5 complex, which provide extra stability to it, consequently leading to the enhanced cellular apoptosis. QC caused a dose dependent increase of DR5 expression in cancer cells but not in normal breast epithelial cells, MCF-10A. QC showed a synergistic effect with TRAIL in causing cancer cell apoptosis. In DR5-KD MCF-10A-Tr (DR5 knocked down) cells, TRAIL+ QC failed to significantly increase the apoptosis but over expression of full length DR5 in DR5-silence cells induced apoptosis, further supporting DR5 as a drug target for QC. An increase in the release of reactive species (ROS and RNS) and activation of enzymes (FADD, CASPASES 3, 8, 9 and cytochrome-C) indicated the involvement of mitochondrial intrinsic pathway in TRAIL+QC mediated apoptosis. In vivo study pointed out that TRAIL+QC co-administration increases the expression of DR5 and reduce the tumor size in xenograft mice. This combined in silico, in vitro and in vivo analysis revealed that QC enhances the cellular apoptosis via the modulation of TRAIL-DR5 complexation and the mitochondrial intrinsic pathway. Impact Journals LLC 2016-08-17 /pmc/articles/PMC5352116/ /pubmed/27542249 http://dx.doi.org/10.18632/oncotarget.11335 Text en Copyright: © 2017 Das et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Paper Das, Sarita Tripathi, Neha Preet, Ranjan Siddharth, Sumit Nayak, Anmada Bharatam, Prasad V. Kundu, Chanakya Nath Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade |
title | Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade |
title_full | Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade |
title_fullStr | Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade |
title_full_unstemmed | Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade |
title_short | Quinacrine induces apoptosis in cancer cells by forming a functional bridge between TRAIL-DR5 complex and modulating the mitochondrial intrinsic cascade |
title_sort | quinacrine induces apoptosis in cancer cells by forming a functional bridge between trail-dr5 complex and modulating the mitochondrial intrinsic cascade |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352116/ https://www.ncbi.nlm.nih.gov/pubmed/27542249 http://dx.doi.org/10.18632/oncotarget.11335 |
work_keys_str_mv | AT dassarita quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade AT tripathineha quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade AT preetranjan quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade AT siddharthsumit quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade AT nayakanmada quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade AT bharatamprasadv quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade AT kunduchanakyanath quinacrineinducesapoptosisincancercellsbyformingafunctionalbridgebetweentraildr5complexandmodulatingthemitochondrialintrinsiccascade |