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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Dove 2019
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.
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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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