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EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+)
BACKGROUND: The role of endothelin receptor type B (EDNRB) isoform 3 involved in Temozolomide (TMZ)-induced melanoma cell death has not yet been elucidated. METHODS: The subcellular localization of EDNRB isoform 3 was determined by confocal and immunoblotting assays. Silencing EDNRB isoform 3 was pe...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689146/ https://www.ncbi.nlm.nih.gov/pubmed/31496797 http://dx.doi.org/10.2147/CMAR.S208604 |
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author | Chen, Yun Shan Liu, Fen Luo, Yi Hong Fan, Yue Xu, Fang Gui Li, Pin Zhou, Bei Pan, Xiu Yu Wang, Chi Chiu Cui, Long |
author_facet | Chen, Yun Shan Liu, Fen Luo, Yi Hong Fan, Yue Xu, Fang Gui Li, Pin Zhou, Bei Pan, Xiu Yu Wang, Chi Chiu Cui, Long |
author_sort | Chen, Yun Shan |
collection | PubMed |
description | BACKGROUND: The role of endothelin receptor type B (EDNRB) isoform 3 involved in Temozolomide (TMZ)-induced melanoma cell death has not yet been elucidated. METHODS: The subcellular localization of EDNRB isoform 3 was determined by confocal and immunoblotting assays. Silencing EDNRB isoform 3 was performed by CRISPR/Cas9. Apoptosis was assessed by annexin V/propium iodide staining and caspases 3/7/9 activity. Mitochondrial membrane potential, reactive oxygen species and mitochondrial Ca(2+) were measured by flow cytometry. Apoptosis protein array was applied. RESULTS: Confocal and immunoblot analyses indicate mitochondrial localization of EDNRB isoform 3 and the first N-terminal (1–22) amino acids are sufficient for its mitochondrial targeting. EDNRB isoform 3 depleted A375 cells significantly confers chemoresistance with mitochondrial depolarization, reduced reactive oxygen species, enhanced mitochondrial Ca(2+) uptake and decreased caspase 9 activation. Additionally, apoptosis array shows that lack of EDNRB isoform 3 has relatively lower expression of phosphorylation of p53 at S392 and a slightly higher expression of Paraoxonase 2. CONCLUSION: Our findings raise the possibility of targeting EDNRB isoform 3 as a new therapeutic strategy in combination with TMZ for melanoma treatment. |
format | Online Article Text |
id | pubmed-6689146 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Dove |
record_format | MEDLINE/PubMed |
spelling | pubmed-66891462019-09-06 EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) Chen, Yun Shan Liu, Fen Luo, Yi Hong Fan, Yue Xu, Fang Gui Li, Pin Zhou, Bei Pan, Xiu Yu Wang, Chi Chiu Cui, Long Cancer Manag Res Original Research BACKGROUND: The role of endothelin receptor type B (EDNRB) isoform 3 involved in Temozolomide (TMZ)-induced melanoma cell death has not yet been elucidated. METHODS: The subcellular localization of EDNRB isoform 3 was determined by confocal and immunoblotting assays. Silencing EDNRB isoform 3 was performed by CRISPR/Cas9. Apoptosis was assessed by annexin V/propium iodide staining and caspases 3/7/9 activity. Mitochondrial membrane potential, reactive oxygen species and mitochondrial Ca(2+) were measured by flow cytometry. Apoptosis protein array was applied. RESULTS: Confocal and immunoblot analyses indicate mitochondrial localization of EDNRB isoform 3 and the first N-terminal (1–22) amino acids are sufficient for its mitochondrial targeting. EDNRB isoform 3 depleted A375 cells significantly confers chemoresistance with mitochondrial depolarization, reduced reactive oxygen species, enhanced mitochondrial Ca(2+) uptake and decreased caspase 9 activation. Additionally, apoptosis array shows that lack of EDNRB isoform 3 has relatively lower expression of phosphorylation of p53 at S392 and a slightly higher expression of Paraoxonase 2. CONCLUSION: Our findings raise the possibility of targeting EDNRB isoform 3 as a new therapeutic strategy in combination with TMZ for melanoma treatment. Dove 2019-08-05 /pmc/articles/PMC6689146/ /pubmed/31496797 http://dx.doi.org/10.2147/CMAR.S208604 Text en © 2019 Chen et al. http://creativecommons.org/licenses/by-nc/3.0/ This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php). |
spellingShingle | Original Research Chen, Yun Shan Liu, Fen Luo, Yi Hong Fan, Yue Xu, Fang Gui Li, Pin Zhou, Bei Pan, Xiu Yu Wang, Chi Chiu Cui, Long EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) |
title | EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) |
title_full | EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) |
title_fullStr | EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) |
title_full_unstemmed | EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) |
title_short | EDNRB isoform 3 confers Temozolomide resistance in A375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial Ca(2+) |
title_sort | ednrb isoform 3 confers temozolomide resistance in a375 melanoma cells by modulating membrane potential, reactive oxygen species and mitochondrial ca(2+) |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6689146/ https://www.ncbi.nlm.nih.gov/pubmed/31496797 http://dx.doi.org/10.2147/CMAR.S208604 |
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