title: Analysis of hypoxia-induced metabolic reprogramming.
abstract: Hypoxia is a common finding in advanced human tumors and is often associated with metastatic dissemination and poor prognosis. Cancer cells adapt to hypoxia by utilizing physiological adaptation pathways that promote a switch from oxidative to glycolytic metabolism. This promotes the conversion of glucose into lactate while limiting its transformation into acetyl coenzyme A (acetyl-CoA). The uptake of glucose and the glycolytic flux are increased under hypoxic conditions, mostly owing to the upregulation of genes encoding glucose transporters and glycolytic enzymes, a process that depends on hypoxia-inducible factor 1 (HIF-1). The reduced delivery of acetyl-CoA to the tricarboxylic acid cycle leads to a switch from glucose to glutamine as the major substrate for fatty acid synthesis in hypoxic cells. In addition, hypoxia induces (1) the HIF-1-dependent expression of BCL2/adenovirus E1B 19-kDa interacting protein 3 (BNIP3) and BNIP3-like (BNIP3L), which trigger mitochondrial autophagy, thereby decreasing the oxidative metabolism of both fatty acids and glucose, and (2) the expression of the sodium-hydrogen exchanger NHE1, which maintains an alkaline intracellular pH. Here, we present a compendium of methods to study hypoxia-induced metabolic alterations.
authors.affiliation: Children's Medical Center Research Institute, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Chendong
authors.initials: C
authors.lastname: Yang
authors.affiliation: Children's Medical Center Research Institute, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Lei
authors.initials: L
authors.lastname: Jiang
authors.affiliation: School of Life Science, University of Science and Technology of China, Hefei, PR China.
authors.forename: Huafeng
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Larissa A
authors.initials: LA
authors.lastname: Shimoda
authors.affiliation: Children's Medical Center Research Institute, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Ralph J
authors.initials: RJ
authors.lastname: DeBerardinis
authors.affiliation: Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; Departments of Pediatrics, Medicine, Oncology, Radiation Oncology, and Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA; McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA. Electronic address: gsemenza@jhmi.edu.
authors.forename: Gregg L
authors.initials: GL
authors.lastname: Semenza
mesh_terms: D001343:Autophagy; D002247:Carbon Isotopes; D018929:Cell Culture Techniques; D003470:Culture Media; D005947:Glucose / Q000032:analysis* / Q000378:metabolism*; D006019:Glycolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D000860:Hypoxia / Q000378:metabolism*; D015151:Immunoblotting; D019344:Lactic Acid / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D008928:Mitochondria / Q000378:metabolism; D015336:Molecular Probe Techniques; D009369:Neoplasms / Q000378:metabolism*; D017382:Reactive Oxygen Species
chemical_list: D002247:Carbon Isotopes; D003470:Culture Media; D017382:Reactive Oxygen Species; D019344:Lactic Acid; D005947:Glucose
keywords: Glycolytic rate; Intracellular pH; Mitochondrial autophagy; Oxygen consumption; Stable isotope labeling; [(13)C]Glucose