title: Glutaminolysis and glycolysis regulation by troglitazone in breast cancer cells: Relationship to mitochondrial membrane potential.
abstract: We studied the roles of glycolysis and glutaminolysis following an acute reduction in mitochondrial membrane potential (Ψ(m)) induced by the thiazolidinedione troglitazone (TRO) and compared the responses with CCCP-induced depolarization in breast cancer derived MCF-7 and MDA-MB-231 cells as well as in the MCF-10A normal breast cell line. TRO and CCCP both acutely reduced Ψ(m) but after 24 h TRO-treated cells had restored Ψ(m) associated with both increased glycolysis and glutaminolysis. In contrast, CCCP-treated cells exhibited only a partial restoration of Ψ(m) associated with increased glycolysis but decreased glutaminolysis. TRO-induced glutaminolysis was coupled to increased ammonium (GDH flux) and decreased alanine production (ALT flux) in all three cell lines. Both cancer cell lines exhibited a higher spontaneous GDH/ALT flux than the normal breast cell line associated with a reduced keto-acid pool. TRO's effect on GDH/ALT fluxes and mitochondrial keto-acid pool homeostasis was additive with glucose withdrawal suggesting limited intramitochondrial pyruvate availability. The TRO-induced acceleration in GDH flux supplies substrate for Complex I contributing to the restoration of Ψ(m) as well as Krebs cycle intermediates for biosynthesis. Inhibiting mitochondrial proton ATPase with oligomycin or nullifying the proton gradient with CCCP prevented both the TRO-induced recovery of Ψ(m) and accelerated GDH flux but restored ALT flux consonant with important roles for proton pumping in regulating GDH flux and Ψ(m) recovery. Blocking enhanced GDH flux reduced DNA synthesis consistent with glutaminolysis via GDH playing an important biosynthetic role in tumorigenesis.
authors.affiliation: Department of Medicine, Feist-Weiller Cancer Center, Louisiana State University Health Science Center, Shreveport, Louisiana 71103, USA. rfrida@lsuhsc.edu
authors.forename: Ellen
authors.initials: E
authors.lastname: Friday
authors.forename: Robert
authors.initials: R
authors.lastname: Oliver
authors.forename: Tomas
authors.initials: T
authors.lastname: Welbourne
authors.forename: Francesco
authors.initials: F
authors.lastname: Turturro
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism*; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D002839:Chromans / Q000494:pharmacology*; D002952:Citric Acid Cycle / Q000502:physiology; D005260:Female; D005947:Glucose / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism*; D006019:Glycolysis / Q000187:drug effects*; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D045162:Thiazolidinediones / Q000494:pharmacology*; D000077288:Troglitazone; D014407:Tumor Cells, Cultured / Q000187:drug effects*
chemical_list: D002839:Chromans; D007004:Hypoglycemic Agents; D045162:Thiazolidinediones; D005973:Glutamine; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D000077288:Troglitazone; D005947:Glucose