Rapid Analysis of Glycolytic and Oxidative Substrate Flux of Cancer Cells in a Microplate
Lisa S. Pike Winer,Min Wu +1 more
TLDR
These rapid, sensitive and high-throughput substrate flux analysis methods introduce highly valuable approaches for developing a greater understanding of genetic and epigenetic pathways that regulate cellular metabolism, and the development of therapies that target cancer metabolism.Abstract:
Cancer cells exhibit remarkable alterations in cellular metabolism, particularly in their nutrient substrate preference. We have devised several experimental methods that rapidly analyze the metabolic substrate flux in cancer cells: glycolysis and the oxidation of major fuel substrates glucose, glutamine, and fatty acids. Using the XF Extracellular Flux analyzer, these methods measure, in real-time, the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of living cells in a microplate as they respond to substrates and metabolic perturbation agents. In proof-of-principle experiments, we analyzed substrate flux and mitochondrial bioenergetics of two human glioblastoma cell lines, SF188s and SF188f, which were derived from the same parental cell line but proliferate at slow and fast rates, respectively. These analyses led to three interesting observations: 1) both cell lines respired effectively with substantial endogenous substrate respiration; 2) SF188f cells underwent a significant shift from glycolytic to oxidative metabolism, along with a high rate of glutamine oxidation relative to SF188s cells; and 3) the mitochondrial proton leak-linked respiration of SF188f cells increased significantly compared to SF188s cells. It is plausible that the proton leak of SF188f cells may play a role in allowing continuous glutamine-fueled anaplerotic TCA cycle flux by partially uncoupling the TCA cycle from oxidative phosphorylation. Taken together, these rapid, sensitive and high-throughput substrate flux analysis methods introduce highly valuable approaches for developing a greater understanding of genetic and epigenetic pathways that regulate cellular metabolism, and the development of therapies that target cancer metabolism.read more
Citations
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Mammary adipocytes stimulate breast cancer invasion through metabolic remodeling of tumor cells
Yuan Yuan Wang,Yuan Yuan Wang,Camille Attané,Delphine Milhas,Béatrice Dirat,Stéphanie Dauvillier,Adrien Guerard,Julia Gilhodes,Ikrame Lazar,Nathalie Alet,Victor Laurent,Sophie Le Gonidec,Denis Biard,Caroline Hervé,Frédéric Bost,Guo Sheng Ren,Françoise Bono,Ghislaine Escourrou,Marc Prentki,Laurence Nieto,Philippe Valet,Catherine Muller +21 more
TL;DR: The results show a complex metabolic symbiosis between tumor-surrounding adipocytes and cancer cells that stimulate their invasiveness, highlighting ATGL as a potential therapeutic target to impede breast cancer progression.
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Metabolic Reprogramming in Glioma
TL;DR: The current state of knowledge regarding the metabolic strategies employed by malignant glioma cells, including aerobic glycolysis; the pentose phosphate pathway; one-carbon metabolism; the tricarboxylic acid cycle, which is central to amino acid metabolism; oxidative phosphorylation; and fatty acid metabolism are presented.
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Tumor cell oxidative metabolism as a barrier to PD-1 blockade immunotherapy in melanoma.
Yana G. Najjar,Ashley V. Menk,Cindy Sander,Uma Rao,Arivarasan Karunamurthy,Roma Bhatia,Shuyan Zhai,John M. Kirkwood,Greg M. Delgoffe +8 more
TL;DR: It is shown that metabolic derangement can vary widely in both degree and type in patient-derived cell lines and in ex vivo analysis of patient samples, such that some cells demonstrate solely deregulated oxidative or glycolytic metabolism.
Journal ArticleDOI
Overcoming Hypoxia by Multistage Nanoparticle Delivery System to Inhibit Mitochondrial Respiration for Photodynamic Therapy
TL;DR: Tumor regression in animal models confirms the feasibility and effectiveness of conquering the PDT‐resistance through abrogating the oxygen consumption, and it is hopeful that such a strategy could shed light on the development of next‐generation PDT‐adjuvant treatment.
Journal ArticleDOI
Determining Maximum Glycolytic Capacity Using Extracellular Flux Measurements.
TL;DR: Maximum glycolytic capacity is experimentally defined, how it differs from glyCOlytic rate is demonstrated, and a protocol for determining it is provided, and the results illustrate the power of extracellular flux analysis to describe the energetics of adherent cells in culture in a fully quantitative way.
References
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Journal ArticleDOI
Understanding the Warburg Effect: The Metabolic Requirements of Cell Proliferation
TL;DR: It is proposed that the metabolism of cancer cells, and indeed all proliferating cells, is adapted to facilitate the uptake and incorporation of nutrients into the biomass needed to produce a new cell.
Journal ArticleDOI
Why do cancers have high aerobic glycolysis
TL;DR: In this article, the authors propose that persistent metabolism of glucose to lactate even in aerobic conditions is an adaptation to intermittent hypoxia in pre-malignant lesions, which leads to microenvironmental acidosis requiring evolution to phenotypes resistant to acid-induced cell toxicity.
Journal ArticleDOI
Mitochondrial formation of reactive oxygen species.
TL;DR: This review describes the main mitochondrial sources of reactive species and the antioxidant defences that evolved to prevent oxidative damage in all the mitochondrial compartments and discusses various physiological and pathological scenarios resulting from an increased steady state concentration of mitochondrial oxidants.