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.2007 Dec 4;104(49):19345-50.
doi: 10.1073/pnas.0709747104. Epub 2007 Nov 21.

Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis

Affiliations

Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis

Ralph J DeBerardinis et al. Proc Natl Acad Sci U S A..

Abstract

Tumor cell proliferation requires rapid synthesis of macromolecules including lipids, proteins, and nucleotides. Many tumor cells exhibit rapid glucose consumption, with most of the glucose-derived carbon being secreted as lactate despite abundant oxygen availability (the Warburg effect). Here, we used 13C NMR spectroscopy to examine the metabolism of glioblastoma cells exhibiting aerobic glycolysis. In these cells, the tricarboxylic acid (TCA) cycle was active but was characterized by an efflux of substrates for use in biosynthetic pathways, particularly fatty acid synthesis. The success of this synthetic activity depends on activation of pathways to generate reductive power (NADPH) and to restore oxaloacetate for continued TCA cycle function (anaplerosis). Surprisingly, both these needs were met by a high rate of glutamine metabolism. First, conversion of glutamine to lactate (glutaminolysis) was rapid enough to produce sufficient NADPH to support fatty acid synthesis. Second, despite substantial mitochondrial pyruvate metabolism, pyruvate carboxylation was suppressed, and anaplerotic oxaloacetate was derived from glutamine. Glutamine catabolism was accompanied by secretion of alanine and ammonia, such that most of the amino groups from glutamine were lost from the cell rather than incorporated into other molecules. These data demonstrate that transformed cells exhibit a high rate of glutamine consumption that cannot be explained by the nitrogen demand imposed by nucleotide synthesis or maintenance of nonessential amino acid pools. Rather, glutamine metabolism provides a carbon source that facilitates the cell's ability to use glucose-derived carbon and TCA cycle intermediates as biosynthetic precursors.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.
Fig. 1.
Glucose metabolism in proliferating glioblastoma cells. (A) The labeling scheme highlights distribution of13C (filled circles) from [1,6-13C2]glucose into various metabolites. Several core pathways, including glycolysis, anaerobic pyruvate metabolism, the TCA cycle, and fatty acid synthesis are shown. (B) Stacked spectra from a real-time [1,6-13C2]glucose perfusion experiment in SF188 cells. Each spectrum represents data summed over 15 min. (C) A partial spectrum from time 6 h highlights enrichment of informative metabolic intermediates. Background from the time 0-h spectrum was subtracted to emphasize the effect of [1,6-13C2]glucose metabolism. (D) Absolute concentrations of [1-13C]glucose, [3-13C]lactate, and [3-13C]alanine during the experiment.
Fig. 2.
Fig. 2.
Tricarboxylic acid cycle activity in glioblastoma cells. (A)The intracellular concentrations of [4-13C]glutamate and [2-13C]glutamate were determined after introduction of [1,6-13C2]glucose. The [4-13C]glutamate results from PDH and the [2-13C]glutamate results from cycling of [4-13C]glutamate. (B) A13C NMR spectrum of a cellular extract from cells cultured in [2-13C]glucose. TheInset is an expansion of the 25- to 60-ppm region highlighting glutamate carbons 2, 3, and 4. Data are from one representative experiment.
Fig. 3.
Fig. 3.
Lipid synthesis in proliferating glioblastoma cells. (A) The labeling of three fatty acyl (FA) resonances, the ω (methyl), ω-2, and methylene [−(CH2)n−] increased throughout the experiment at a constant rate, demonstrating continuous transfer of13C from glucose into fatty acids. (B) Cells were cultured in medium containing [U-14C6]glucose, and then lipids were extracted and analyzed by thin-layer chromatography. The average and standard deviation for three parallel cultures are shown for each lipid species. (C) Cells were cultured in medium containing 10 mM [U-13C6]glucose, and then lipids were extracted and analyzed by NMR spectroscopy. TheInset shows an expansion of the 23-ppm region, highlighting the ω-1 multiplet (d, doublet; t, triplet). (D) Cells were cultured in medium containing 4 mM [3-13C]glutamine and 10 mM unlabeled glucose (time 0–6.25 h), followed by medium containing 4 mM [3-13C]glutamine and 10 mM [1,6-13C2]glucose (time 6.25–13 h). The relative rate (R.R.) for the increase in −(13CH2)n− signal was calculated for both halves of the experiment.
Fig. 4.
Fig. 4.
Glutamine metabolism in proliferating glioblastoma cells. (A) The labeling scheme begins with [3-13C]glutamine (blue) at the top right of the mitochondrion. For simplicity, extramitochondrial glutamine metabolism and aspartate production are not shown. The pathway outlines glutaminolysis, the conversion of glutamine-derived carbon to lactate by using malic enzyme (ME), which produces NADPH. The appearance of [2-13C]Ac-CoA (red) is from the addition of [1,6-13C2]glucose. (B) Design for two-stage perfusion experiment. In the first stage, cells received [3-13C]glutamine and unlabeled glucose as a bolus and then as a continuous feed. In the second stage, cells received [3-13C]glutamine and [1,6-13C2]glucose. (C) Stacked spectra acquired during the two-stage perfusion experiment. Each spectrum represents data summed over 15 min. (D) Intracellular concentrations of [3-13C]glutamate, [2-13C]glutamate, [4-13C]glutamate, and [3-13C]aspartate during both stages. (E) Concentration of [2-13C]lactate and [3-13C]lactate during perfusion with [3-13C]glutamine.
Fig. 5.
Fig. 5.
Glutamine is the major anaplerotic substrate in proliferating glioblastoma cells. (A) Stacked spectra of the 34.3-ppm region ([4-13C]glutamate) obtained during a perfusion with [1,6-13C2]glucose (Left, from Fig. 1B) and during the two-stage perfusion (Right). (B) Intracellular concentration of glutamate isotopomers labeled at C-4. “Total” concentration equals the sum of [4-13C]glutamate and [3,4-13C2]glutamate. (C) Cumulative production of extracellular15NH3 and [15N]alanine from cells cultured withl-[α-15N]glutamine. This experiment was performed three times, and one representative time course is shown.
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References

    1. Cooper EH, Barkhan P, Hale AJ. Br J Haematol. 1963;9:101–111. - PubMed
    1. Hedeskov CJ. Biochem J. 1968;110:373–380. - PMC - PubMed
    1. Edinger AL, Thompson CB. Mol Biol Cell. 2002;13:2276–2288. - PMC - PubMed
    1. Warburg O. Klin Wochenschr Berl. 1925;4:534–536.
    1. Wang T, Marquardt C, Foker J. Nature. 1976;261:702–705. - PubMed

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