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ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency

Previously, the in vitro growth of cancer stem cells in the form of tumor spheres from five different brain cancer cell lines was found to be methionine-dependent. As this earlier work indicated that ALDH1L2, a folate-dependent mitochondria aldehyde dehydrogenase gene, is upregulated in glioblastoma...

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Autores principales: Quéré, Maëlle, Alberto, Jean-Marc, Broly, Franck, Hergalant, Sébastien, Christov, Christo, Gauchotte, Guillaume, Guéant, Jean-Louis, Namour, Farès, Battaglia-Hsu, Shyue-Fang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105572/
https://www.ncbi.nlm.nih.gov/pubmed/35565854
http://dx.doi.org/10.3390/nu14091887
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author Quéré, Maëlle
Alberto, Jean-Marc
Broly, Franck
Hergalant, Sébastien
Christov, Christo
Gauchotte, Guillaume
Guéant, Jean-Louis
Namour, Farès
Battaglia-Hsu, Shyue-Fang
author_facet Quéré, Maëlle
Alberto, Jean-Marc
Broly, Franck
Hergalant, Sébastien
Christov, Christo
Gauchotte, Guillaume
Guéant, Jean-Louis
Namour, Farès
Battaglia-Hsu, Shyue-Fang
author_sort Quéré, Maëlle
collection PubMed
description Previously, the in vitro growth of cancer stem cells in the form of tumor spheres from five different brain cancer cell lines was found to be methionine-dependent. As this earlier work indicated that ALDH1L2, a folate-dependent mitochondria aldehyde dehydrogenase gene, is upregulated in glioblastoma stem cells, we invalidated this gene using CRISPR-cas 9 technique in this present work. We reported here that this invalidation was effective in U251 glioblastoma cells, and no cas9 off target site could be detected by genome sequencing of the two independent knockout targeting either exon I or exon III. The knockout of ALDH1L2 gene in U251 cells rendered the growth of the cancer stem cells of U251 methionine independent. In addition, a much higher ROS (reactive oxygen radicals) level can be detected in the knockout cells compared to the wild type cells. Our evidence here linked the excessive ROS level of the knockout cells to reduced total cellular NADPH. Our evidence suggested also that the cause of the slower growth of the knockout turmor sphere may be related to its partial differentiation.
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spelling pubmed-91055722022-05-14 ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency Quéré, Maëlle Alberto, Jean-Marc Broly, Franck Hergalant, Sébastien Christov, Christo Gauchotte, Guillaume Guéant, Jean-Louis Namour, Farès Battaglia-Hsu, Shyue-Fang Nutrients Article Previously, the in vitro growth of cancer stem cells in the form of tumor spheres from five different brain cancer cell lines was found to be methionine-dependent. As this earlier work indicated that ALDH1L2, a folate-dependent mitochondria aldehyde dehydrogenase gene, is upregulated in glioblastoma stem cells, we invalidated this gene using CRISPR-cas 9 technique in this present work. We reported here that this invalidation was effective in U251 glioblastoma cells, and no cas9 off target site could be detected by genome sequencing of the two independent knockout targeting either exon I or exon III. The knockout of ALDH1L2 gene in U251 cells rendered the growth of the cancer stem cells of U251 methionine independent. In addition, a much higher ROS (reactive oxygen radicals) level can be detected in the knockout cells compared to the wild type cells. Our evidence here linked the excessive ROS level of the knockout cells to reduced total cellular NADPH. Our evidence suggested also that the cause of the slower growth of the knockout turmor sphere may be related to its partial differentiation. MDPI 2022-04-30 /pmc/articles/PMC9105572/ /pubmed/35565854 http://dx.doi.org/10.3390/nu14091887 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
Quéré, Maëlle
Alberto, Jean-Marc
Broly, Franck
Hergalant, Sébastien
Christov, Christo
Gauchotte, Guillaume
Guéant, Jean-Louis
Namour, Farès
Battaglia-Hsu, Shyue-Fang
ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency
title ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency
title_full ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency
title_fullStr ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency
title_full_unstemmed ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency
title_short ALDH1L2 Knockout in U251 Glioblastoma Cells Reduces Tumor Sphere Formation by Increasing Oxidative Stress and Suppressing Methionine Dependency
title_sort aldh1l2 knockout in u251 glioblastoma cells reduces tumor sphere formation by increasing oxidative stress and suppressing methionine dependency
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105572/
https://www.ncbi.nlm.nih.gov/pubmed/35565854
http://dx.doi.org/10.3390/nu14091887
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