Energy metabolism transition in multi-cellular human tumor spheroids.
It is thought that glycolysis is the predominant energy pathway in cancer, particularly in solid and poorly vascularized tumors where hypoxic regions develop. To evaluate whether glycolysis does effectively predominate for ATP supply and to identify the underlying biochemical mechanisms, the glycolytic and oxidative phosphorylation (OxPhos) fluxes, ATP/ADP ratio, phosphorylation potential, and expression and activity of relevant energy metabolism enzymes were determined in multi-cellular tumor spheroids, as a model of human solid tumors. In HeLa and Hek293 young-spheroids, the OxPhos flux and cytochrome c oxidase protein content and activity were similar to those observed in monolayer cultured cells, whereas the glycolytic flux increased two- to fourfold; the contribution of OxPhos to ATP supply was 60%. In contrast, in old-spheroids, OxPhos, ATP content, ATP/ADP ratio, and phosphorylation potential diminished 50-70%, as well as the activity (88%) and content (3 times) of cytochrome c oxidase. Glycolysis and hexokinase increased significantly (both, 4 times); consequently glycolysis was the predominant pathway for ATP supply (80%). These changes were associated with an increase (3.3 times) in the HIF-1alpha content. After chronic exposure, both oxidative and glycolytic inhibitors blocked spheroid growth, although the glycolytic inhibitors, 2-deoxyglucose and gossypol (IC(50) of 15-17 nM), were more potent than the mitochondrial inhibitors, casiopeina II-gly, laherradurin, and rhodamine 123 (IC(50) > 100 nM). These results suggest that glycolysis and OxPhos might be considered as metabolic targets to diminish cellular proliferation in poorly vascularized, hypoxic solid tumors.
Rodríguez-Enríquez
Sara
S
Departamento de Bioquímica, Instituto Nacional de Cardiología, Mexico, Mexico. saren960104@hotmail.com
Gallardo-Pérez
Juan Carlos
JC
Avilés-Salas
Alejandro
A
Marín-Hernández
Alvaro
A
Carreño-Fuentes
Liliana
L
Maldonado-Lagunas
Vilma
V
Moreno-Sánchez
Rafael
R
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D003847:Deoxyglucose / Q000378:metabolism; D004734:Energy Metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006019:Glycolysis / Q000502:physiology; D006072:Gossypol / Q000378:metabolism; D006801:Humans; D007783:Lactones / Q000378:metabolism; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology; D009942:Organometallic Compounds / Q000378:metabolism; D010085:Oxidative Phosphorylation; D010100:Oxygen / Q000378:metabolism; D020112:Rhodamine 123; D018874:Spheroids, Cellular
D005456:Fluorescent Dyes; D007783:Lactones; D009942:Organometallic Compounds; C505569:casiopeina II-glycine; C432547:laherradurin; D020112:Rhodamine 123; D000255:Adenosine Triphosphate; D003847:Deoxyglucose; D006072:Gossypol; D010100:Oxygen