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The Warburg effect: a balance of flux analysis

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Abstract

Cancer metabolism is characterized by increased macromolecular syntheses through coordinated increases in energy and substrate metabolism. The observation that cancer cells produce lactate in an environment of oxygen sufficiency (aerobic glycolysis) is a central theme of cancer metabolism known as the Warburg effect. Aerobic glycolysis in cancer metabolism is accompanied by increased pentose cycle and anaplerotic activities producing energy and substrates for macromolecular synthesis. How these processes are coordinated is poorly understood. Recent advances have focused on molecular regulation of cancer metabolism by oncogenes and tumor suppressor genes which regulate numerous enzymatic steps of central glucose metabolism. In the past decade, new insights in cancer metabolism have emerged through the application of stable isotopes particularly from 13C carbon tracing. Such studies have provided new evidence for system-wide changes in cancer metabolism in response to chemotherapy. Interestingly, experiments using metabolic inhibitors on individual biochemical pathways all demonstrate similar system-wide effects on cancer metabolism as in targeted therapies. Since biochemical reactions in the Warburg effect place competing demands on available precursors, high energy phosphates and reducing equivalents, the cancer metabolic system must fulfill the condition of balance of flux (homeostasis). In this review, the functions of the pentose cycle and of the tricarboxylic acid (TCA) cycle in cancer metabolism are analyzed from the balance of flux point of view. Anticancer treatments that target molecular signaling pathways or inhibit metabolism alter the invasive or proliferative behavior of the cancer cells by their effects on the balance of flux (homeostasis) of the cancer metabolic phenotype.

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Notes

  1. In a cellular system, biochemical reactions are constrained by physical and chemical factors such as compartmentalization or tunneling; Km and Vmax of Michaelis–Menten kinetics; cofactor concentrations, and conservation of mass and energy (balance of flux).

  2. The two types of reactions of Eq. 1 and 2 are either energy consuming or energy producing reaction. There is a large group of reversible reactions in which energy state of the system is not affected.

  3. The concept of balance of flux is applicable to reactions in one functional or physical compartment, as well as in all the compartments of the cellular metabolic network as a whole.

  4. Glucose is the ultimate source of energy and carbon for amino acids and fatty acid synthesis. As a general rule in metabolism, energy is conserved (stored) in the form of fatty acids, and carbon source is conserved or regenerated through anaplerosis. Thus, glucose is generally not used fot ATP production, which is consistent with the occurrence of hypoglycemia in fatty acid oxidation abnormality and the development of diabetes under high fat dietary condition.

  5. Futile cycle refers to a pair of reversible reactions in which the net result is the consumption of ATP or NADPH. A typical example of a pair of reactions is the combined action of glucokinase and glucose-6-phosphatase. It should be noted that these reactions do not take place contemporaneously, and reaction in either direction potentially affects its “downstream” reactions.

  6. [1, 2-13C2]-OAA is converted to [1, 2-13C2]-glutamate and [3, 4-13C2]-OAA to [3-13C]-glutamate. The carbon 2–5 fragment has only singly labeled mass isotopomer from recycling.

  7. The relation between m1/m2 ratio (r) and relative anaplerotic flux (Y) is given by the equation:

    $${\text{Y}} = \left( { 1- {\text{r}}} \right)/{\text{r}}$$

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Acknowledgments

This work was supported by the National Institutes of Health (P01AT003960) and the Hirshberg Foundation for Pancreatic Cancer Research, V.B.P was supported by DK58132-01A2 and NIAID Grant U19AI091175-01.

Conflict of interest

The authors have no conflicts of interest to disclose.

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This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Department of Medicine, Diabetes Center, Stable Isotope and Metabolomics Core Facility, Albert Einstein College of Medicine Diabetes Center, Bronx, NY, USA

    B. Vaitheesvaran

  2. Department of Pathology, University of Southern California, Los Angeles, CA, USA

    J. Xu

  3. Division of Endocrinology and Metabolism, Department of Pediatrics, University of California, Los Angeles, CA, USA

    J. Yee & W.-N. Lee

  4. Department of Medicine, University of California, Los Angeles, CA, USA

    Q.-Y. Lu & V. L. Go

  5. Functional Genomics/Proteomics Laboratories, Creighton University Medical Center, Omaha, NE, USA

    G. G. Xiao

  6. School of Pharmaceutical Science and Technology at Dalian University of Technology, Dalian, China

    G. G. Xiao

  7. LA Biomedical Research Institute, Torrance, CA, USA

    W.-N. Lee

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  1. B. Vaitheesvaran
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  2. J. Xu
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  3. J. Yee
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  6. G. G. Xiao
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Correspondence to W.-N. Lee.

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11306_2014_760_MOESM1_ESM.jpg

Supplemental Figure S1: Glycolytic/gluconeogenic pathways and their regulation. The regulation of glycolysis is accomplished through a series of futile cycles (shown as double arrows) and reversible reactions (shown as single bidirectional arrows). The direction of flux is regulated by multiple connected pathways. In addition, it is controlled by expression and/or activation of enzymes. Since glucose is needed for numerous substrate syntheses, multiple regulatory points (double arrows) are necessary to achieve fine control of glucose flux and its directions. HEX stands for hexokinase; G6Pase, glucose-6-phosphatase; GS glycogen synthase; GP, glycogenphosphorylase; PFK, phosphofructose kinase; GAPDH, glyceraldehyde 3 phosphate dehydrogenase; PGK, phosphoglycerate kinase; PGM, phosphoglycerate mutase; ENO, enolase; PKM, pyruvate kinase isozymes M1/M2 and LDH, lactate dehydrogenase. These key control points are regulated by oncogenes and cancer suppressor genes (D’Alessandro and Zolla 2012). (JPEG 228 kb)

11306_2014_760_MOESM2_ESM.jpg

Supplemental Figure S2: Balance of flux equations for anaplerotic reactions. These reactions allow communication (substrate exchange) between cytosolic and mitochondrial compartments. Not included in the OAA or malate balance are equations for transamination, PEPCK, maleic reaction. Together with reactions from Figure 5, these reactions form the basis for the balance of flux model of TCA cycle compartment. (JPEG 332 kb)

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Vaitheesvaran, B., Xu, J., Yee, J. et al. The Warburg effect: a balance of flux analysis. Metabolomics 11, 787–796 (2015). https://doi.org/10.1007/s11306-014-0760-9

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