title: Increased concentrations of fructose 2,6-bisphosphate contribute to the Warburg effect in phosphatase and tensin homolog (PTEN)-deficient cells.
abstract: Unlike normal differentiated cells, tumor cells metabolize glucose via glycolysis under aerobic conditions, a hallmark of cancer known as the Warburg effect. Cells lacking the commonly mutated tumor suppressor PTEN exhibit a glycolytic phenotype reminiscent of the Warburg effect. This has been traditionally attributed to the hyperactivation of PI3K/Akt signaling that results from PTEN loss. Here, we propose a novel mechanism whereby the loss of PTEN negatively affects the activity of the E3 ligase APC/C-Cdh1, resulting in the stabilization of the enzyme PFKFB3 and increased synthesis of its product fructose 2,6-bisphosphate (F2,6P2). We discovered that when compared with wild-type cells, PTEN knock-out mouse embryonic fibroblasts (PTEN KO MEF) have 2-3-fold higher concentrations of F2,6P2, the most potent allosteric activator of the glycolytic enzyme phosphofructokinase-1 (PFK-1). Reintroduction of either wild-type or phosphatase mutant PTEN in the PTEN KO cells effectively lowers F2,6P2 to the wild-type levels and reduces their lactate production. PTEN KO cells were found to have high protein levels of PFKFB3, which directly contribute to the increased concentrations of F2,6P2. PTEN enhances interaction between PFKFB3 and Cdh1, and overexpression of Cdh1 down-regulates the PFKFB3 protein level in wild-type, but not in PTEN-deficient cells. Importantly, we found that the degradation of endogenous PFKFB3 in PTEN KO cells occurs at a slower rate than in wild-type cells. Our results suggest an important role for F2,6P2 in the metabolic reprogramming of PTEN-deficient cells that has important consequences for cell proliferation.
authors.affiliation: From the Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
authors.forename: Lucía
authors.initials: L
authors.lastname: Cordero-Espinoza
authors.forename: Thilo
authors.initials: T
authors.lastname: Hagen
mesh_terms: D064173:Anaphase-Promoting Complex-Cyclosome / Q000378:metabolism; D000818:Animals; D064200:Cdh1 Proteins / Q000378:metabolism; D002460:Cell Line; D005635:Fructosediphosphates / Q000378:metabolism*; D055786:Gene Knockout Techniques; D006019:Glycolysis; D051379:Mice; D051059:PTEN Phosphohydrolase / Q000172:deficiency* / Q000235:genetics; D025481:Phosphofructokinase-2 / Q000378:metabolism; D059748:Proteolysis
chemical_list: D064200:Cdh1 Proteins; D005635:Fructosediphosphates; C577585:Fzr1 protein, mouse; C027652:fructose 2,6-diphosphate; D064173:Anaphase-Promoting Complex-Cyclosome; D025481:Phosphofructokinase-2; D051059:PTEN Phosphohydrolase
keywords: APC/C-Cdh1; E3 Ubiquitin Ligase; Fructose 2,6-Bisphosphate; Glycolysis; PFKFB3; Phosphofructokinase; Pten