Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells

American Journal of Physiology - Cell Physiology - Tập 292 Số 1 - Trang C125-C136 - 2007
Min Wu1, Andy Neilson, Amy L. Swift, Rebecca Moran, James Tamagnine, Diane Parslow, Suzanne Armistead, Kristie Lemire, Jim Orrell, Jay Teich, Steve Chomicz, David A. Ferrick
1Seahorse Bioscience, 16 Esquire Road, North Billerica, MA 01862, USA

Tóm tắt

Increased conversion of glucose to lactic acid associated with decreased mitochondrial respiration is a unique feature of tumors first described by Otto Warburg in the 1920s. Recent evidence suggests that the Warburg effect is caused by oncogenes and is an underlying mechanism of malignant transformation. Using a novel approach to measure cellular metabolic rates in vitro, the bioenergetic basis of this increased glycolysis and reduced mitochondrial respiration was investigated in two human cancer cell lines, H460 and A549. The bioenergetic phenotype was analyzed by measuring cellular respiration, glycolysis rate, and ATP turnover of the cells in response to various pharmacological modulators. H460 and A549 cells displayed a dependency on glycolysis and an ability to significantly upregulate this pathway when their respiration was inhibited. The converse, however, was not true. The cell lines were attenuated in oxidative phosphorylation (OXPHOS) capacity and were unable to sufficiently upregulate mitochondrial OXPHOS when glycolysis was disabled. This observed mitochondrial impairment was intimately linked to the increased dependency on glycolysis. Furthermore, it was demonstrated that H460 cells were more glycolytic, having a greater impairment of mitochondrial respiration, compared with A549 cells. Finally, the upregulation of glycolysis in response to mitochondrial ATP synthesis inhibition was dependent on AMP-activated protein kinase activity. In summary, our results demonstrate a bioenergetic phenotype of these two cancer cell lines characterized by increased rate of glycolysis and a linked attenuation in their OXPHOS capacity. These metabolic alterations provide a mechanistic explanation for the growth advantage and apoptotic resistance of tumor cells.

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Tài liệu tham khảo

Almeida A, Almeida J, Bolanos JP, Moncada S. Different responses of astrocytes and neurons to nitric oxide: the role of glycolytically generated ATP in astrocyte protection. Proc Natl Acad Sci USA 98: 15294–15299, 2001.

Almeida A, Moncada S, Bolanos JP. Nitric oxide switches on glycolysis through the AMP protein kinase and 6-phosphofructo-2-kinase pathway. Nat Cell Biol 6: 45–51, 2004.

Blum R, Jacob-Hirsch J, Amariglio N, Rechavi G, Kloog Y. Ras inhibition in glioblastoma down-regulates hypoxia-inducible factor-1alpha, causing glycolysis shutdown and cell death. Cancer Res 65: 999–1006, 2005.

Boren J, Cascante M, Marin S, Comin-Anduix B, Centelles JJ, Lim S, Bassilian S, Ahmed S, Lee WN, Boros LG. Gleevec (STI571) influences metabolic enzyme activities and glucose carbon flow toward nucleic acid and fatty acid synthesis in myeloid tumor cells. J Biol Chem 276: 37747–3753, 2001.

Brand KA, Hermfisse U. Aerobic glycolysis by proliferating cells: a protective strategy against reactive oxygen species. FASEB J 11: 388–395, 1997.

Brand MD. The efficiency and plasticity of mitochondrial energy transduction. Biochem Soc Trans 33: 897–904, 2005.

Clark LC Jr. Monitor and control of blood and tissue oxygen tension. Trans Am Soc Artif Intern Organs 2: 41–48, 1956.

Cuezva JM, Krajewska M, de Heredia ML, Krajewski S, Santamaria G, Kim H, Zapata JM, Marusawa H, Chamorro M, Reed JC. The bioenergetic signature of cancer: a marker of tumor progression. Cancer Res 62: 6674–6681, 2002.

Cuezva JM, Chen G, Alonso AM, Isidoro A, Misek DE, Hanash SM, Beer DG. The bioenergetic signature of lung adenocarcinomas is a molecular marker of cancer diagnosis and prognosis. Carcinogenesis 25: 1157–1163, 2004.

Dang CV, Semenza GL. Oncogenic alterations of metabolism. Trends Biochem Sci 24: 68–72, 1999.

Elstrom RL, Bauer DE, Buzzai M, Karnauskas R, Harris MH, Plas DR, Zhuang H, Cinalli RM, Alavi A, Rudin CM, Thompson CB. Akt stimulates aerobic glycolysis in cancer cells. Cancer Res 64: 3892–3899, 2004.

Fantin VR, St-Pierre J, Leder P. Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell 9: 425–434, 2006.

Gambhir SS. Molecular imaging of cancer with positron emission tomography. Nat Rev Cancer 2: 683–693, 2002.

Gatenby RA, Gillies RJ. Why do cancers have high aerobic glycolysis? Nat Rev Cancer 4: 891–899, 2004.

Gottschalk S, Anderson N, Hainz C, Eckhardt SG, Serkova NJ. Imatinib (STI571)-mediated changes in glucose metabolism in human leukemia BCR-ABL-positive cells. Clin Cancer Res 10: 6661–6618, 2004.

Hardie DG, Scott JW, Pan DA, Hudson ER. Management of cellular energy by the AMP-activated protein kinase system. FEBS Lett 546: 113–120, 2003.

Hatzivassiliou G, Andreadis C, Thompson CB. Akt-directed metabolic alterations in cancer. Drug Discov Today Dis Mech 2: 255–262, 2005.

Herst PM, Tan AS, Scarlett DJ, Berridge MV. Cell surface oxygen consumption by mitochondrial gene knockout cells. Biochim Biophys Acta 1656: 79–87, 2004.

Isidoro A, Casado E, Redondo A, Acebo P, Espinosa E, Alonso AM, Cejas P, Hardisson D, Fresno Vara JA, Belda-Iniesta C, Gonzalez-Baron M, Cuezva JM. Breast carcinomas fulfill the Warburg hypothesis and provide metabolic markers of cancer prognosis. Carcinogenesis 26: 2095–2104, 2005.

Kelloff GJ, Krohn KA, Larson SM, Weissleder R, Mankoff DA, Hoffman JM, Link JM, Guyton KZ, Eckelman WC, Scher HI, O'Shaughnessy J, Cheson BD, Sigman CC, Tatum JL, Mills GQ, Sullivan DC, Woodcock J. The progress and promise of molecular imaging probes in oncologic drug development. Clin Cancer Res 11: 2785–2808, 2005.

Kim J, Gardner LB, Dang CV. Oncogenic alterations of metabolism and the Warburg effect. Drug Discov Today Dis Mech 2: 233–238, 2005.

Koukourakis MI, Giatromanolaki A, Simopoulos C, Polychronidis A, Sivridis E. Lactate dehydrogenase 5 (LDH5) relates to up-regulated hypoxia inducible factor pathway and metastasis in colorectal cancer. Clin Exp Metastasis 22:25–30, 2005.

Kraus-Berthier L, Jan M, Guilbaud N, Naze M, Pierre A, Atassi G. Histology and sensitivity to anticancer drugs of two human non-small cell lung carcinomas implanted in the pleural cavity of nude mice. Clin Cancer Res 6: 297–304. 2000.

Maity A, Tuttle SW. 2-Deoxyglucose and radiosensitization: Teaching an old DOG new tricks? Cancer Biol Ther 5: 824–826, 2006.

Matoba S, Kang JG, Patino WD, Wragg A, Boehm M, Gavrilova O, Hurley PJ, Bunz F, Hwang PM. p53 regulates mitochondrial respiration. Science 312: 1650–1653, 2006.

Mazurek S, Eigenbrodt E. The tumor metabolome. Anticancer Res 23: 1149–1154, 2003.

Nicholls DG, Ward MW. Mitochondrial membrane potential and neuronal glutamate excitotoxicity: mortality and millivolts. Trends Neurosci 23: 166–174, 2000.

Owicki JC, Parce JW. Biosensors based on the energy metabolism of living cells: the physical chemistry and cell biology of extracellular acidification. Biosens Bioelectron 7: 255–272, 1992.

Parce JW, Owicki JC, Kercso KM, Sigal GB, Wada HG, Muir VC, Bousse LJ, Ross KL, Sikic BI, McConnell HM. Detection of cell-affecting agents with a silicon biosensor. Science 246: 243–247, 1989.

Ramanathan A, Wang C, Scheiber SL. Perturbational profiling of a cell-line model of tumorigenesis by using metabolic measurements. Proc Natl Acad Sci USA 102: 5992–5997, 2005.

Ristow M. Oxidative metabolism in cancer growth. Curr Opin Clin Nutr Metab Care 9: 339–345, 2006.

Robey RB, Hay N. Akt, hexokinase, mTOR: Targeting cellular energy metabolism for cancer therapy. Drug Discov Today Dis Mech 2: 239–246, 2005.

Rolfe DF, Brand MD. The physiological significance of mitochondrial proton leak in animal cells and tissues. Biosci Rep 17: 9–16, 1997.

Rolfe DF, Brown GC. Cellular energy utilization and molecular origin of standard metabolic rate in mammals. Physiol Rev 77: 731–758, 1997.

Rossignol R, Gilkerson R, Aggeler R, Yamagata K, Remington SJ, Capaldi RA. Energy substrate modulates mitochondrial structure and oxidative capacity in cancer cells. Cancer Res 64: 985–993, 2004.

Santamaria G, Martinez-Diez M, Fabregat I, Cuezva JM. Efficient execution of cell death in non-glycolytic cells requires the generation of ROS controlled by the activity of mitochondrial H+-ATP synthase. Carcinogenesis 27: 925–935, 2006.

Schulz TJ, Thierbach R, Voigt A, Drewes G, Mietzner B, Steinberg P, Pfeiffer AF, Ristow M. Induction of oxidative metabolism by mitochondrial frataxin inhibits cancer growth: Otto Warburg revisited. J Biol Chem 281: 977–981, 2006.

Shaw RJ, Kosmatka M, Bardeesy N, Hurley RL, Witters LA, DePinho RA, Cantley LC. The tumor suppressor LKB1 kinase directly activates AMP-activated kinase and regulates apoptosis in response to energy stress. Proc Natl Acad Sci USA 101: 3329–3335, 2004.

Simonnet H, Alazard N, Pfeiffer K, Gallou C, Beroud C, Demont J, Bouvier R, Schagger H, Godinot C. Low mitochondrial respiratory chain content correlates with tumor aggressiveness in renal cell carcinoma. Carcinogenesis 23: 759–768, 2002.

Simonnet H, Demont J, Pfeiffer K, Guenaneche L, Bouvier R, Brandt U, Schagger H, Godinot C. Mitochondrial complex I is deficient in renal oncocytomas. Carcinogenesis 24: 1461–1466, 2003.

Tan MG, Ooi LL, Aw SE, Hui KM. Cloning and identification of hepatocellular carcinoma down-regulated mitochondrial carrier protein, a novel liver-specific uncoupling protein. J Biol Chem 279: 45235–4544, 2004.

Warburg O. On the origin of cancer cells. Science 123: 309–314, 1956.

Warburg O, Posener K, Negelein E. Uber den Stoffwechsel der Carcinomzelle. Biochem Z 152: 309–344, 1924.

Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk-Melody J, Wu M, Ventre J, Doebber T, Fujii N, Musi N, Hirshman MF, Goodyear LJ, Moller DE. Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest 108: 1167–1174, 2001.