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Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model

Continuous activation of hypoxia pathways in pheochromocytomas and paragangliomas (PPGLs) is associated with higher disease aggressiveness, for which effective treatment strategies are still missing. Most of the commonly used in vitro models lack characteristics of these pseudohypoxic tumors, includ...

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Autores principales: Helm, Jana, Drukewitz, Stephan, Poser, Isabel, Richter, Susan, Friedemann, Markus, William, Doreen, Mohr, Hermine, Nölting, Svenja, Robledo, Mercedes, Bornstein, Stefan R., Eisenhofer, Graeme, Bechmann, Nicole
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8834104/
https://www.ncbi.nlm.nih.gov/pubmed/35159368
http://dx.doi.org/10.3390/cells11030560
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author Helm, Jana
Drukewitz, Stephan
Poser, Isabel
Richter, Susan
Friedemann, Markus
William, Doreen
Mohr, Hermine
Nölting, Svenja
Robledo, Mercedes
Bornstein, Stefan R.
Eisenhofer, Graeme
Bechmann, Nicole
author_facet Helm, Jana
Drukewitz, Stephan
Poser, Isabel
Richter, Susan
Friedemann, Markus
William, Doreen
Mohr, Hermine
Nölting, Svenja
Robledo, Mercedes
Bornstein, Stefan R.
Eisenhofer, Graeme
Bechmann, Nicole
author_sort Helm, Jana
collection PubMed
description Continuous activation of hypoxia pathways in pheochromocytomas and paragangliomas (PPGLs) is associated with higher disease aggressiveness, for which effective treatment strategies are still missing. Most of the commonly used in vitro models lack characteristics of these pseudohypoxic tumors, including elevated expression of hypoxia-inducible factor (HIF) 2α. To address this shortcoming, we investigated whether recurrent hypoxia cycles lead to continuous activation of hypoxia pathways under normoxic conditions and whether this pseudohypoxia is associated with increased cellular aggressiveness. Rat pheochromocytoma cells (PC12) were incubated under hypoxia for 24 h every 3–4 days, up to 20 hypoxia–reoxygenation cycles, resulting in PC12 Z20 cells. PC12 Z20 control cells were obtained by synchronous cultivation under normoxia. RNA sequencing revealed upregulation of HIF2α in PC12 Z20 cells and a pseudohypoxic gene signature that overlapped with the gene signature of pseudohypoxic PPGLs. PC12 Z20 cells showed a higher growth rate, and the migration and adhesion capacity were significantly increased compared with control cells. Changes in global methylation, together with the pseudohypoxic conditions, may be responsible for the increased aggressiveness of this new model. The established sub-cell line with characteristics of pseudohypoxic PPGLs represent a complementary model for further investigations, for example, with regard to new therapeutic approaches.
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spelling pubmed-88341042022-02-12 Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model Helm, Jana Drukewitz, Stephan Poser, Isabel Richter, Susan Friedemann, Markus William, Doreen Mohr, Hermine Nölting, Svenja Robledo, Mercedes Bornstein, Stefan R. Eisenhofer, Graeme Bechmann, Nicole Cells Article Continuous activation of hypoxia pathways in pheochromocytomas and paragangliomas (PPGLs) is associated with higher disease aggressiveness, for which effective treatment strategies are still missing. Most of the commonly used in vitro models lack characteristics of these pseudohypoxic tumors, including elevated expression of hypoxia-inducible factor (HIF) 2α. To address this shortcoming, we investigated whether recurrent hypoxia cycles lead to continuous activation of hypoxia pathways under normoxic conditions and whether this pseudohypoxia is associated with increased cellular aggressiveness. Rat pheochromocytoma cells (PC12) were incubated under hypoxia for 24 h every 3–4 days, up to 20 hypoxia–reoxygenation cycles, resulting in PC12 Z20 cells. PC12 Z20 control cells were obtained by synchronous cultivation under normoxia. RNA sequencing revealed upregulation of HIF2α in PC12 Z20 cells and a pseudohypoxic gene signature that overlapped with the gene signature of pseudohypoxic PPGLs. PC12 Z20 cells showed a higher growth rate, and the migration and adhesion capacity were significantly increased compared with control cells. Changes in global methylation, together with the pseudohypoxic conditions, may be responsible for the increased aggressiveness of this new model. The established sub-cell line with characteristics of pseudohypoxic PPGLs represent a complementary model for further investigations, for example, with regard to new therapeutic approaches. MDPI 2022-02-05 /pmc/articles/PMC8834104/ /pubmed/35159368 http://dx.doi.org/10.3390/cells11030560 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Helm, Jana
Drukewitz, Stephan
Poser, Isabel
Richter, Susan
Friedemann, Markus
William, Doreen
Mohr, Hermine
Nölting, Svenja
Robledo, Mercedes
Bornstein, Stefan R.
Eisenhofer, Graeme
Bechmann, Nicole
Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model
title Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model
title_full Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model
title_fullStr Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model
title_full_unstemmed Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model
title_short Treatment of Pheochromocytoma Cells with Recurrent Cycles of Hypoxia: A New Pseudohypoxic In Vitro Model
title_sort treatment of pheochromocytoma cells with recurrent cycles of hypoxia: a new pseudohypoxic in vitro model
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8834104/
https://www.ncbi.nlm.nih.gov/pubmed/35159368
http://dx.doi.org/10.3390/cells11030560
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