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Simultaneous steady-state and dynamic13C NMR can differentiate alternative routes of pyruvate metabolism in living cancer cells
Chendong Yang; Crystal Harrison; Eunsook S. Jin; David T. Chuang; A. Dean Sherry; Craig R. Malloy; Matthew E. Merritt; Ralph J. DeBerardinis(Profiled Authors: David T Chuang; Ralph J DeBerardinis; Eunsook S Jin; Craig R Malloy; Matthew E Merritt; Dean Sherry)
Journal of Biological Chemistry. 2014;289(9):6212-6224.Abstract
Metabolic reprogramming facilitates cancer cell growth, so quantitative metabolic flux measurements could produce useful biomarkers. However, current methods to analyze flux in vivo provide either a steady-state overview of relative activities (infusion of 13C and analysis of extracted metabolites) or a dynamic view of a few reactions (hyperpolarized 13C spectroscopy). Moreover, although hyperpolarization has successfully quantified pyruvate-lactate exchanges, its ability to assess mitochondrial pyruvate metabolism is unproven in cancer. Here, we combined 13C hyperpolarization and isotopomer analysis to quantify multiple fates of pyruvate simultaneously. Two cancer cell lines with divergent pyruvate metabolism were incubated with thermally polarized [3-13C]pyruvate for several hours, then briefly exposed to hyperpolarized [1-13C]pyruvate during acquisition of NMR spectra using selective excitation to maximize detection of H[13C]O
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