title: Phosphofructokinase 2 and glycolysis in HT29 human colon adenocarcinoma cell line. Regulation by insulin and phorbol esters.
abstract: Kinetic properties of phosphofructokinase 2 (PFK2) and regulation of glycolysis by phorbol 12-myristate 13-acetate (PMA) and insulin were investigated in highly glycolytic HT29 colon cancer cells. PFK2 was found to be inhibited by citrate and, to a lesser extent, by phosphoenolpyruvate and ADP, but to be insensitive to inhibition by sn-glycerol phosphate. From these kinetic data, PFK2 from HT29 cells appears different from the liver form, but resembles somewhat the heart isoenzyme. Fructose 2,6-bisphosphate (Fru-2,6-P2) levels, glucose consumption and lactate production are increased in a dose-dependent manner in HT29 cells treated with PMA or insulin. The increase in Fru-2,6-P2 can be related to an increase in the Vmax. of PFK2, persisting after the enzyme has been precipitated with poly(ethylene glycol), without change in the Km for fructose 6-phosphate. The most striking effects of PMA and insulin on Fru-2,6-P2 production are observed after long-term treatment (24 h) and are abolished by actinomycin, cycloheximide and puromycin, suggesting that protein synthesis is involved. Furthermore, the effects of insulin and PMA on glucose consumption, lactate production, Fru-2,6-P2 levels and PFK2 activity are additive, and the effect of insulin on Fru-2,6-P2 production is not altered by pre-treatment of the cells with the phorbol ester. This suggests that these effects are exerted by separate mechanisms.
authors.affiliation: INSERM Unité 317, Institut de Physiologie, Université Paul Sabatier, Toulouse, France.
authors.forename: C
authors.initials: C
authors.lastname: Denis-Pouxviel
authors.forename: T
authors.initials: T
authors.lastname: Gauthier
authors.forename: D
authors.initials: D
authors.lastname: Daviaud
authors.forename: J C
authors.initials: JC
authors.lastname: Murat
mesh_terms: D000230:Adenocarcinoma / Q000201:enzymology*; D003110:Colonic Neoplasms / Q000201:enzymology*; D006019:Glycolysis; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007700:Kinetics; D007773:Lactates / Q000378:metabolism; D025481:Phosphofructokinase-2; D010770:Phosphotransferases / Q000096:biosynthesis*; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D007328:Insulin; D007773:Lactates; D010770:Phosphotransferases; D025481:Phosphofructokinase-2; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate