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Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate

BACKGROUND: Cell growth requires fatty acids for membrane synthesis. Fatty acids are assembled from 2-carbon units in the form of acetyl-CoA (AcCoA). In nutrient and oxygen replete conditions, acetyl-CoA is predominantly derived from glucose. In hypoxia, however, flux from glucose to acetyl-CoA decr...

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Autores principales: Kamphorst, Jurre J, Chung, Michelle K, Fan, Jing, Rabinowitz, Joshua D
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4322440/
https://www.ncbi.nlm.nih.gov/pubmed/25671109
http://dx.doi.org/10.1186/2049-3002-2-23
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author Kamphorst, Jurre J
Chung, Michelle K
Fan, Jing
Rabinowitz, Joshua D
author_facet Kamphorst, Jurre J
Chung, Michelle K
Fan, Jing
Rabinowitz, Joshua D
author_sort Kamphorst, Jurre J
collection PubMed
description BACKGROUND: Cell growth requires fatty acids for membrane synthesis. Fatty acids are assembled from 2-carbon units in the form of acetyl-CoA (AcCoA). In nutrient and oxygen replete conditions, acetyl-CoA is predominantly derived from glucose. In hypoxia, however, flux from glucose to acetyl-CoA decreases, and the fractional contribution of glutamine to acetyl-CoA increases. The significance of other acetyl-CoA sources, however, has not been rigorously evaluated. Here we investigate quantitatively, using (13)C-tracers and mass spectrometry, the sources of acetyl-CoA in hypoxia. RESULTS: In normoxic conditions, cultured cells produced more than 90% of acetyl-CoA from glucose and glutamine-derived carbon. In hypoxic cells, this contribution dropped, ranging across cell lines from 50% to 80%. Thus, under hypoxia, one or more additional substrates significantly contribute to acetyl-CoA production. (13)C-tracer experiments revealed that neither amino acids nor fatty acids are the primary source of this acetyl-CoA. Instead, the main additional source is acetate. A large contribution from acetate occurs despite it being present in the medium at a low concentration (50–500 μM). CONCLUSIONS: Acetate is an important source of acetyl-CoA in hypoxia. Inhibition of acetate metabolism may impair tumor growth. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2049-3002-2-23) contains supplementary material, which is available to authorized users.
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spelling pubmed-43224402015-02-11 Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate Kamphorst, Jurre J Chung, Michelle K Fan, Jing Rabinowitz, Joshua D Cancer Metab Research BACKGROUND: Cell growth requires fatty acids for membrane synthesis. Fatty acids are assembled from 2-carbon units in the form of acetyl-CoA (AcCoA). In nutrient and oxygen replete conditions, acetyl-CoA is predominantly derived from glucose. In hypoxia, however, flux from glucose to acetyl-CoA decreases, and the fractional contribution of glutamine to acetyl-CoA increases. The significance of other acetyl-CoA sources, however, has not been rigorously evaluated. Here we investigate quantitatively, using (13)C-tracers and mass spectrometry, the sources of acetyl-CoA in hypoxia. RESULTS: In normoxic conditions, cultured cells produced more than 90% of acetyl-CoA from glucose and glutamine-derived carbon. In hypoxic cells, this contribution dropped, ranging across cell lines from 50% to 80%. Thus, under hypoxia, one or more additional substrates significantly contribute to acetyl-CoA production. (13)C-tracer experiments revealed that neither amino acids nor fatty acids are the primary source of this acetyl-CoA. Instead, the main additional source is acetate. A large contribution from acetate occurs despite it being present in the medium at a low concentration (50–500 μM). CONCLUSIONS: Acetate is an important source of acetyl-CoA in hypoxia. Inhibition of acetate metabolism may impair tumor growth. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/2049-3002-2-23) contains supplementary material, which is available to authorized users. BioMed Central 2014-12-11 /pmc/articles/PMC4322440/ /pubmed/25671109 http://dx.doi.org/10.1186/2049-3002-2-23 Text en © Kamphorst et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Kamphorst, Jurre J
Chung, Michelle K
Fan, Jing
Rabinowitz, Joshua D
Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate
title Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate
title_full Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate
title_fullStr Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate
title_full_unstemmed Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate
title_short Quantitative analysis of acetyl-CoA production in hypoxic cancer cells reveals substantial contribution from acetate
title_sort quantitative analysis of acetyl-coa production in hypoxic cancer cells reveals substantial contribution from acetate
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4322440/
https://www.ncbi.nlm.nih.gov/pubmed/25671109
http://dx.doi.org/10.1186/2049-3002-2-23
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