Özcan, S. C.Fernandez, Y. I.Muchut, R. J.Iglesias, A. A.Gürpınar, Y.Clem, A. L.Chesney, J. A.Yalçın, A.2024-02-272024-02-272020-05-27Özcan, S. C. vd. (2020). "PFKFB2 regulates glycolysis and proliferation in pancreatic cancer cells". Molecular and Cellular Biochemistry, 470(1-2), 115-129.0300-81771573-4919https://link.springer.com/article/10.1007/s11010-020-03751-5https://hdl.handle.net/11452/39991Tumor cells increase glucose metabolism through glycolysis and pentose phosphate pathways to meet the bioenergetic and biosynthetic demands of rapid cell proliferation. The family of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB1-4) are key regulators of glucose metabolism via their synthesis of fructose-2,6-bisphosphate (F2,6BP), a potent activator of glycolysis. Previous studies have reported the co-expression of PFKFB isozymes, as well as the mRNA splice variants of particular PFKFB isozymes, suggesting non-redundant functions. Majority of the evidence demonstrating a requirement for PFKFB activity in increased glycolysis and oncogenic properties in tumor cells comes from studies on PFKFB3 and PFKFB4 isozymes. In this study, we show that the PFKFB2 isozyme is expressed in tumor cell lines of various origin, overexpressed and localizes to the nucleus in pancreatic adenocarcinoma, relative to normal pancreatic tissue. We then demonstrate the differential intracellular localization of two PFKFB2 mRNA splice variants and that, when ectopically expressed, cytoplasmically localized mRNA splice variant causes a greater increase in F2,6BP which coincides with an increased glucose uptake, as compared with the mRNA splice variant localizing to the nucleus. We then show that PFKFB2 expression is required for steady-state F2,6BP levels, glycolytic activity, and proliferation of pancreatic adenocarcinoma cells. In conclusion, this study may provide a rationale for detailed investigation of PFKFB2's requirement for the glycolytic and oncogenic phenotype of pancreatic adenocarcinoma cells.eninfo:eu-repo/semantics/closedAccessPancreatic adenocarcinomaGlycolysisPFKFB2Fructose-26-bisphosphate6-phosphofructo-2-kinase PFKFB36-phosphofructo-2-kinase/fructose-2,6-BisphosphataseExpressionMetabolismGlucoseMigraitonInvasionCell biologyAdenocarcinomaCell differentiationCell line, tumorCell nucleusCell ProliferationCytoplasmGene expression regulation, enzymologicGene expression regulation, neoplasticGene SilencingGlycolysisHeLa CellsHumansIsoenzymesPancreasPancreatic neoplasmsPhenotypePhosphofructokinase-2RNA splicingRNA, messengerArticle2-s2.0-850846661151151294701-232415418https://doi.org/10.1007/s11010-020-03751-5Cell biologyPhosphofructokinase-2; Apoptosis; GlycolysisGlucoseMessenger RNAPfkfb2 proteinProteinSmall interfering RNAUnclassified drug6 phosphofructo 2 kinaseIsoenzymeMessenger RNAPFKFB2 protein, humanA-375 cell lineA-549 cell lineAmino acid sequenceArticleBxPC-3 cell lineCell nucleusCell proliferationCellular distributionColony formationComparative studyControlled studyCytosolDU145 cell lineEctopic expressionFemaleGenetic transfectionGlucose intakeGlucose metabolismGlucose transportGlycolysisHCT 116 cell lineHeLa cell lineHumanHuman cellImmunofluorescence testImmunoreactivityNuclear localization signalANC-1 cell lineProtein expression levelPancreas adenocarcinomaProtein expressionReal time polymerase chain reactionTransient expressionUpregulationUterine cervix carcinomaWestern blottingWound healing assayAdenocarcinomaCell differentiationCell proliferationCytoplasmEnzymologyGene expression regulationGene silencingGeneticsMetabolismPancreasPancreas tumorPathologyPhenotypeRNA splicingTumor cell linePhysiology