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Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway
In this study, we addressed how silibinin enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in various cancer cells. Combined treatment with silibinin and TRAIL (silibinin/TRAIL) induced apoptosis accompanied by the activation of caspase-3, caspase-8, caspas...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828202/ https://www.ncbi.nlm.nih.gov/pubmed/29535810 http://dx.doi.org/10.18632/oncotarget.23129 |
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author | Dilshara, Matharage Gayani Jayasooriya, Rajapaksha Gedara Prasad Tharanga Molagoda, Ilandarage Menu Neelaka Jeong, Jin-Woo Lee, Seungheon Park, Sang Rul Kim, Gi-Young Choi, Yung Hyun |
author_facet | Dilshara, Matharage Gayani Jayasooriya, Rajapaksha Gedara Prasad Tharanga Molagoda, Ilandarage Menu Neelaka Jeong, Jin-Woo Lee, Seungheon Park, Sang Rul Kim, Gi-Young Choi, Yung Hyun |
author_sort | Dilshara, Matharage Gayani |
collection | PubMed |
description | In this study, we addressed how silibinin enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in various cancer cells. Combined treatment with silibinin and TRAIL (silibinin/TRAIL) induced apoptosis accompanied by the activation of caspase-3, caspase-8, caspase-9, and Bax, and cytosolic accumulation of cytochrome c. Anti-apoptotic proteins such as Bcl-2, IAP-1, and IAP-2 were inhibited as well. Silibinin also triggered TRAIL-induced apoptosis in A549 cells through upregulation of death receptor 5 (DR5). Pretreatment with DR5/Fc chimeric protein and DR5-targeted small interfering RNA (siRNA) significantly blocked silibinin/TRAIL-mediated apoptosis in A549 cells. Furthermore, silibinin increased the production of reactive oxygen species (ROS), which led to the induction of TRAIL-mediated apoptosis through DR5 upregulation. Antioxidants such as N-acetyl-L-cysteine and glutathione reversed the apoptosis-inducing effects of TRAIL. Silibinin further induced endoplasmic reticulum (ER) stress as was indicated by the increase in ER marker proteins such as PERK, eIF2α, and ATF-4, which stimulate the expression of CCAAT/enhancer binding protein homologous protein (CHOP). CHOP-targeted siRNA eliminated the induction of DR5 and resulted in a significant decrease in silibinin/TRAIL-mediated apoptosis. We also found that silibinin/TRAIL-induced apoptosis was accompanied with intracellular influx of Ca(2+), which was stimulated by ER stress and the Ca(2+) chelator, ethylene glycol tetraacetic acid (EGTA). Ca(2+)/calmodulin-dependent protein kinase (CaMKII) inhibitor, K252a, blocked silibinin/TRAIL-induced DR5 expression along with TRAIL-mediated apoptosis. Accordingly, we showed that ROS/ER stress-induced CaMKII activated Sp1, which is an important transcription factor for DR5 expression. Our results showed that silibinin enhanced TRAIL-induced apoptosis by upregulating DR5 expression through the ROS-ER stress-CaMKII-Sp1 axis. |
format | Online Article Text |
id | pubmed-5828202 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-58282022018-03-13 Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway Dilshara, Matharage Gayani Jayasooriya, Rajapaksha Gedara Prasad Tharanga Molagoda, Ilandarage Menu Neelaka Jeong, Jin-Woo Lee, Seungheon Park, Sang Rul Kim, Gi-Young Choi, Yung Hyun Oncotarget Research Paper In this study, we addressed how silibinin enhances tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in various cancer cells. Combined treatment with silibinin and TRAIL (silibinin/TRAIL) induced apoptosis accompanied by the activation of caspase-3, caspase-8, caspase-9, and Bax, and cytosolic accumulation of cytochrome c. Anti-apoptotic proteins such as Bcl-2, IAP-1, and IAP-2 were inhibited as well. Silibinin also triggered TRAIL-induced apoptosis in A549 cells through upregulation of death receptor 5 (DR5). Pretreatment with DR5/Fc chimeric protein and DR5-targeted small interfering RNA (siRNA) significantly blocked silibinin/TRAIL-mediated apoptosis in A549 cells. Furthermore, silibinin increased the production of reactive oxygen species (ROS), which led to the induction of TRAIL-mediated apoptosis through DR5 upregulation. Antioxidants such as N-acetyl-L-cysteine and glutathione reversed the apoptosis-inducing effects of TRAIL. Silibinin further induced endoplasmic reticulum (ER) stress as was indicated by the increase in ER marker proteins such as PERK, eIF2α, and ATF-4, which stimulate the expression of CCAAT/enhancer binding protein homologous protein (CHOP). CHOP-targeted siRNA eliminated the induction of DR5 and resulted in a significant decrease in silibinin/TRAIL-mediated apoptosis. We also found that silibinin/TRAIL-induced apoptosis was accompanied with intracellular influx of Ca(2+), which was stimulated by ER stress and the Ca(2+) chelator, ethylene glycol tetraacetic acid (EGTA). Ca(2+)/calmodulin-dependent protein kinase (CaMKII) inhibitor, K252a, blocked silibinin/TRAIL-induced DR5 expression along with TRAIL-mediated apoptosis. Accordingly, we showed that ROS/ER stress-induced CaMKII activated Sp1, which is an important transcription factor for DR5 expression. Our results showed that silibinin enhanced TRAIL-induced apoptosis by upregulating DR5 expression through the ROS-ER stress-CaMKII-Sp1 axis. Impact Journals LLC 2017-12-07 /pmc/articles/PMC5828202/ /pubmed/29535810 http://dx.doi.org/10.18632/oncotarget.23129 Text en Copyright: © 2018 Dilshara et al. http://creativecommons.org/licenses/by/3.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/) (CC-BY), which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Research Paper Dilshara, Matharage Gayani Jayasooriya, Rajapaksha Gedara Prasad Tharanga Molagoda, Ilandarage Menu Neelaka Jeong, Jin-Woo Lee, Seungheon Park, Sang Rul Kim, Gi-Young Choi, Yung Hyun Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway |
title | Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway |
title_full | Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway |
title_fullStr | Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway |
title_full_unstemmed | Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway |
title_short | Silibinin sensitizes TRAIL-mediated apoptosis by upregulating DR5 through ROS-induced endoplasmic reticulum stress-Ca(2+)-CaMKII-Sp1 pathway |
title_sort | silibinin sensitizes trail-mediated apoptosis by upregulating dr5 through ros-induced endoplasmic reticulum stress-ca(2+)-camkii-sp1 pathway |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5828202/ https://www.ncbi.nlm.nih.gov/pubmed/29535810 http://dx.doi.org/10.18632/oncotarget.23129 |
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