| 2013 |
PFKFB3 controls endothelial cell filopodia/lamellipodia formation and directional migration by compartmentalizing with F-actin in motile protrusions, and overrides the pro-stalk activity of Notch signaling to regulate vessel branching tip/stalk cell competition. |
Loss-of-function (siRNA/genetic KO), mosaic in vitro and in vivo sprouting assays, subcellular co-localization with F-actin, epistasis with Notch pathway |
Cell |
High |
23911327
|
| 2002 |
PFKFB3 (iPFK-2) is induced by hypoxia in cancer cells and its expression and fructose-2,6-bisphosphate levels increase specifically during S phase of the cell cycle, functionally coupling glycolytic activation to cell cycle progression. |
In situ hybridization, immunohistochemistry, cell-cycle fractionation, hypoxia treatment of cultured cells |
Cancer research |
Medium |
12384552
|
| 2015 |
During mitotic arrest, AMPK directly phosphorylates PFKFB3, increasing glycolysis to replace lost oxidative phosphorylation (caused by mitophagy-dependent mitochondrial loss), thereby promoting cell survival. PFKFB3 protein levels also increase due to mitotic-specific translational activation of its mRNA. |
AMPK kinase assay, phospho-specific immunoblotting, siRNA knockdown, autophagy induction/inhibition, breast cancer cell viability assays |
Nature cell biology |
High |
26322680
|
| 2020 |
The lncRNA AGPG binds to and stabilizes PFKFB3 protein by preventing APC/C-mediated ubiquitination, thereby protecting PFKFB3 from proteasomal degradation and leading to accumulation of PFKFB3 and enhanced glycolytic flux in cancer cells. |
Co-immunoprecipitation, ubiquitination assay, proteasome inhibition, siRNA/overexpression, PDX tumor models |
Nature communications |
High |
32198345
|
| 2018 |
Cisplatin induces acetylation of PFKFB3 at lysine 472 (K472), which impairs its nuclear localization signal (NLS) activity, causing cytoplasmic accumulation of PFKFB3. Cytoplasmic PFKFB3 is then phosphorylated by AMPK, leading to PFKFB3 activation and enhanced glycolysis that protects cells from DNA damage-induced apoptosis. |
Site-directed mutagenesis (K472 acetylation-dead mutant), subcellular fractionation, AMPK phosphorylation assay, siRNA knockdown, xenograft model |
Nature communications |
High |
29410405
|
| 2014 |
PFKFB3 promotes cell cycle progression and suppresses apoptosis via its product F2,6BP activating Cdk1, which phosphorylates p27 (T187) leading to p27 ubiquitination and proteasomal degradation. siRNA silencing of PFKFB3 inhibits Cdk1 activity, stabilizes p27, and causes G1/S arrest; co-silencing of p27 reverses these effects. |
siRNA silencing, co-siRNA epistasis, Cdk1 kinase assay, p27 protein stability/ubiquitination assay, flow cytometry (cell cycle/apoptosis), HeLa cells |
Cell death & disease |
High |
25032860
|
| 2012 |
NMDAR activation stabilizes PFKFB3 protein in neurons (normally degraded by APC/C-Cdh1 proteasomal pathway via its KEN motif) and promotes PFKFB3 release from the nucleus to cytosol, switching neuronal metabolism from pentose-phosphate pathway (PPP) to glycolysis, causing oxidative stress and apoptotic neuronal death. |
NMDAR stimulation in cortical neurons, APC/C-Cdh1 KEN-motif deletion mutant, subcellular fractionation, PPP/glycolysis flux measurements, overexpression of G6PD rescue experiment |
Cell death and differentiation |
High |
22421967
|
| 2018 |
PFKFB3 rapidly relocates into ionizing radiation-induced nuclear foci in an MRN-ATM-γH2AX-MDC1-dependent manner and is critical for recruitment of homologous recombination (HR) repair proteins and HR activity. PFKFB3 enzymatic activity is required for ribonucleotide reductase M2 (RRM2) recruitment, deoxynucleotide incorporation during DNA repair, and maintenance of dNTP levels. |
Immunofluorescence co-localization with DNA damage/HR foci, pharmacological inhibitor (KAN0438757), siRNA, HR reporter assay, dNTP level measurements, radiosensitization assays in transformed vs. non-transformed cells |
Nature communications |
High |
30250201
|
| 2006 |
siRNA silencing of PFKFB3 in HeLa cells decreases fructose-2,6-bisphosphate, lactate, and ATP, leading to reduced cell viability, cell cycle delay, increased apoptosis, and inhibition of anchorage-independent growth. |
siRNA knockdown, metabolite measurement (F2,6BP, lactate, ATP), flow cytometry (cell cycle, apoptosis), soft agar colony formation |
FEBS letters |
Medium |
16698023
|
| 2020 |
CPEB4 binds cytoplasmic polyadenylation elements (CPEs) in the 3'-UTR of PFKFB3 mRNA to promote its polyadenylation and translational upregulation (not transcriptional), thereby increasing PFKFB3 protein and glycolysis during hepatic stellate cell activation and liver fibrosis. |
CPEB4 siRNA knockdown, CPEB4-KO mice, RIP/pulldown of PFKFB3 mRNA by CPEB4, mRNA polyadenylation assay, PFKFB3 protein levels, primary HSC and LX2 cells |
Gastroenterology |
High |
32169429
|
| 2020 |
c-Src phosphorylates PFKFB3 at tyrosine 194 (Tyr194), activating PFKFB3 and stimulating glycolysis. PFKFB3-Y194F knockin mice show impaired glycolysis and attenuated spontaneous colon tumor formation when crossed with APCmin/+ mice. |
In vitro kinase assay (c-Src + PFKFB3), site-directed mutagenesis (Y194F), PFKFB3-Y194F knockin mice, APCmin/+ genetic cross, cell proliferation and xenograft assays, clinical tumor sample correlation |
Cell reports |
High |
32209481
|
| 2014 |
Estradiol (E2) promotes PFKFB3 mRNA transcription through direct estrogen receptor (ER) binding to the PFKFB3 promoter, leading to increased PFKFB3 protein, elevated F2,6BP, and enhanced glucose uptake and glycolysis in ER+ breast cancer cells. |
ER-chromatin binding (promoter assay), siRNA, PFKFB3 inhibitor, 14C-glucose uptake, F2,6BP measurement, apoptosis assay in MCF-7 cells |
The Journal of biological chemistry |
Medium |
24515104
|
| 2019 |
Autophagy machinery, specifically the UBA domain of p62/sequestosome-1, physically interacts with PFKFB3; autophagy reduces PFKFB3 expression and induces cellular dormancy, whereas impairment of autophagy (knockdown of Atg3, Atg7, or p62) restores PFKFB3 expression and reactivates proliferation in dormant breast cancer stem cells. |
Co-immunoprecipitation (PFKFB3-p62 interaction), siRNA (Atg3, Atg7, p62), microarray, cell dormancy/proliferation assays, in vivo metastasis model |
Nature communications |
High |
31413316
|
| 2018 |
Nuclear PFKFB3 silencing in hepatocellular carcinoma decreases AKT phosphorylation and reduces ERCC1 expression, impairing DNA repair and causing G2/M arrest and apoptosis, revealing a non-glycolytic nuclear function. |
siRNA knockdown, AKT phosphorylation western blot, ERCC1 expression, cell cycle analysis, xenograft model |
Cell death & disease |
Medium |
29559632
|
| 2019 |
PFKFB3-driven glycolysis in pulmonary endothelial cells promotes pulmonary hypertension; endothelial PFKFB3 knockout reduces HIF2A levels, decreases production of growth factors (PDGFB, FGF2) and proinflammatory factors (CXCL12, IL-1β), and prevents vascular smooth muscle cell proliferation and leukocyte recruitment. |
Conditional endothelial Pfkfb3-KO mice (constitutive and inducible), heterozygous global KO, 3PO pharmacological inhibition, Western blot, immunostaining of lung ECs |
Proceedings of the National Academy of Sciences |
High |
31213542
|
| 2024 |
PFKFB3-mediated tubular glycolysis increases kidney lactate levels, which drives histone lactylation (particularly H4K12la) at the promoters of NF-κB pathway genes (Ikbkb, Rela, Relb), activating their transcription and promoting renal inflammation and fibrosis. PFKFB3 also directly activates IKKβ, IκBα, and p65. |
PTC-specific PFKFB3-KO mice, ChIP-seq (H4K12la at gene promoters), kidney IRI model, heterozygous KO and pharmacological inhibition, metabolite measurement (lactate), Western blot |
Kidney international |
High |
38789037
|
| 2022 |
PFKFB3-driven glycolysis in endothelial cells drives EndoMT by hijacking glucose flux from the pentose phosphate pathway, reducing cytoplasmic NADPH production. Mitochondrial NADPH efflux via the isocitrate/α-ketoglutarate shuttle replenishes cytoplasmic NADPH but impairs mitochondrial respiration by hampering iron-sulfur cluster biosynthesis. |
PFKFB3 haplodeficiency and overexpression, NADPH flux measurement, mitochondrial respiration assay (Seahorse), iron-sulfur cluster biosynthesis analysis, pharmacological inhibition with salvianolic acid C |
Signal transduction and targeted therapy |
Medium |
36045132
|
| 2019 |
PFKFB3 inhibition by small molecule AZ67 prevents NMDAR excitotoxicity-induced NADPH oxidation, redox stress, and apoptotic neuronal death; in vivo administration of AZ67 alleviates motor discoordination and brain infarct in middle carotid artery occlusion ischemia/reperfusion model in mice. |
Pharmacological inhibition (AZ67), NADPH measurement, apoptosis assay in primary neurons, oxygen-glucose deprivation model, in vivo MCAO mouse model |
Scientific reports |
Medium |
31406177
|
| 2022 |
In CLN7 neuronal ceroid lipofuscinosis, failure of autophagy causes mitochondrial accumulation and elevated mitochondrial ROS (mROS), which signal to stabilize PFKFB3 (normally proteasomally degraded in healthy neurons) via a mROS-dependent protein stabilization cascade, driving aberrant glycolysis in neurons and contributing to disease pathogenesis. |
Cln7∆ex2 mouse model, in vivo genetic mROS measurement, PFKFB3 protein stability assays, AZ67 inhibitor treatment in vivo and in CLN7 patient-derived cells |
Nature communications |
High |
35087090
|
| 2022 |
PFKFB3 interacts with cyclin-dependent kinase 4 (CDK4) in the nucleus of kidney tubular cells following cisplatin treatment, leading to CDK4 activation and consequent phosphorylation/inactivation of the retinoblastoma tumor suppressor (Rb), causing apoptosis; this is independent of PFKFB3's canonical glycolytic function. |
Co-immunoprecipitation (PFKFB3-CDK4), CDK4 kinase activity assay, Rb phosphorylation immunoblot, renal proximal tubule-specific PFKFB3-KO mice, CDK4 inhibition rescue |
Translational research |
High |
36243313
|
| 2023 |
OTUD4 deubiquitinase binds PFKFB3 and blocks its ubiquitination-mediated proteasomal degradation, stabilizing PFKFB3 protein in cardiac fibroblasts stimulated with TGF-β1 and promoting glycolysis-driven fibroblast activation. |
Co-immunoprecipitation (PFKFB3-OTUD4), ubiquitination assay, siRNA knockdown, cardiac fibroblast activation assays, post-MI mouse model |
Journal of molecular medicine |
Medium |
37162556
|
| 2022 |
ROCK2 stabilizes PFKFB3 protein in osteosarcoma cells by modifying its ubiquitination and reducing proteasomal degradation; PFKFB3 is epistatic to ROCK2 in promoting proliferation and metastasis, as PFKFB3 overexpression rescues ROCK2 knockdown-induced growth impairment. |
siRNA knockdown, PFKFB3 overexpression rescue, ubiquitination assay, proliferation/invasion assays, clinical tissue correlation |
Experimental cell research |
Medium |
31678169
|
| 2022 |
CYLD stabilizes p53 and promotes its nuclear translocation by removing K63- and K48-linked ubiquitin chains from p53, enabling p53 to bind the PFKFB3 promoter and inhibit its transcription. CYLD also interacts with FZR1 to promote APC/C-FZR1 E3 ligase activity, which ubiquitinates and degrades PFKFB3 via 26S proteasomal system. |
Co-immunoprecipitation (CYLD-FZR1, CYLD-p53), ubiquitination assay (K48/K63 chains on p53), ChIP (p53 binding to PFKFB3 promoter), proteasome inhibition, siRNA |
Cancer letters |
Medium |
35131382
|
| 2020 |
Fascin promotes PFKFB3 transcription by activating YAP1 through its canonical actin-bundling activity; YAP1 binds a TEAD1/4 binding motif 30 bp upstream of the PFKFB3 transcription start site to increase PFKFB3 expression and glycolysis in lung cancer cells. |
ChIP (YAP1-TEAD at PFKFB3 promoter), fascin actin-bundling mutant, YAP1 siRNA, PFKFB3 promoter luciferase, xenograft metastasis models, organoid cultures |
Cancer letters |
High |
34303764
|
| 2021 |
YAP acts as a transcriptional co-activator with TEAD1 to bind the PFKFB3 promoter and increase PFKFB3 expression under hypoxia in endothelial cells, thereby promoting glycolysis and angiogenesis; YAP silencing inhibits endothelial glycolysis and can be rescued by enforced PFKFB3 expression. |
ChIP (YAP-TEAD1 binding to PFKFB3 promoter), siRNA, PFKFB3 overexpression rescue, Seahorse glycolysis assay, intravitreal siRNA injection in CNV/OIR mouse models |
Angiogenesis |
Medium |
33400016
|
| 2022 |
PFKFB3-driven glycolysis in macrophages activates HIF-1α/HIF-2α and NF-κB, inducing M1/M2 polarization markers and pro-angiogenic cytokines, thereby reprogramming macrophages toward an angiogenic phenotype that promotes choroidal neovascularization; myeloid-specific PFKFB3 KO attenuates this process. |
Myeloid cell-specific PFKFB3 KO mice, bone marrow-derived macrophage stimulation, PFKFB3 inhibitor (AZ67), HIF-1α/HIF-2α and NF-κB pathway immunoblotting, laser-induced CNV model |
British journal of pharmacology |
High |
35830274
|
| 2022 |
PFKFB3-driven glycolysis is required for actin polymerization in macrophages; homozygous loss of Pfkfb3 impairs macrophage efferocytosis by disrupting actin-dependent cytoskeletal function, exacerbating atherosclerosis. |
Homozygous Pfkfb3-KO Apoe-/- mice, efferocytosis assay, actin polymerization measurement, flow cytometry |
British journal of pharmacology |
Medium |
35834356
|
| 2022 |
ALK fusion oncoproteins promote PFKFB3 transcription through the downstream transcription factor STAT3; pharmacological or genetic blockade of ALK reduces PFKFB3 expression and glycolysis, and PFKFB3 inhibition can overcome TKI resistance in ALK-mutant cancer cells. |
Quantitative proteomics, STAT3 ChIP/reporter assay, ALK TKI treatment, PFKFB3 siRNA and overexpression, correlation in clinical ALK+ NSCLC samples |
Oncogene |
Medium |
36064579
|
| 2022 |
The deubiquitinase UCHL1 stabilizes PFKFB3 in astrocytes by cleaving K48-linked ubiquitin chains; the UCHL1/PFKFB3 axis increases lactate production, which drives histone H4K8 lactylation (H4K8la), subsequently activating transcription of Uchl1 and glycolysis genes, forming a positive feedback loop that supports astrocytic metabolic reprogramming after spinal cord injury. |
Genetic deletion of Uchl1 and Pfkfb3, K48-ubiquitin chain cleavage assay, ChIP (H4K8la at gene promoters), lactate measurement, in vivo SCI model, scRNA-seq analysis |
Cell death and differentiation |
Medium |
40016338
|
| 2023 |
In pulmonary arterial smooth muscle cells, HIF-1α binds the promoter of miR-26a-5p to inhibit its expression; miR-26a-5p directly targets PFKFB3 (validated by dual-luciferase assay), and PFKFB3 enhances phosphorylation of ULK1 to promote autophagy and cell proliferation in PAH. |
ChIP (HIF-1α at miR-26a-5p promoter), dual-luciferase reporter assay (miR-26a-5p targeting PFKFB3 3'UTR), ULK1 phosphorylation immunoblot, tandem mRFP-GFP-LC3B autophagy assay, adeno-miR-26a-5p in vivo rat PAH model |
FASEB journal |
Medium |
37052859
|
| 2003 |
The human PFKFB3 gene spans 32.5 kb on chromosome 10p15.3-p15.2, contains 19 exons (15 normally expressed), and encodes a 590 amino acid, 66.9 kDa bifunctional protein with the highest kinase/phosphatase activity ratio among all PFKFB isozymes. |
Genomic sequencing, exon-intron junction determination, FISH chromosomal localization, open reading frame analysis |
International journal of oncology |
Medium |
12963966
|
| 2018 |
X-ray crystal structures of PFKFB3 were used to guide structure-based design of N-aryl 6-aminoquinoxaline inhibitors that bind the kinase active site; the most potent compound showed IC50 of 14 nM for PFKFB3 kinase and 0.49 μM for F2,6BP production in HCT116 cells. |
X-ray crystallography, docking, structure-activity relationship (SAR), enzymatic IC50 assay, cellular F2,6BP assay |
ChemMedChem |
High |
30378281
|
| 2022 |
LINC00930 lncRNA acts as a scaffold to recruit the RBBP5 and GCN5 complex to the PFKFB3 promoter, increasing H3K4 trimethylation and H3K9 acetylation at the PFKFB3 promoter to epigenetically activate PFKFB3 transcription and glycolysis in nasopharyngeal carcinoma. |
ChIP (H3K4me3, H3K9ac at PFKFB3 promoter), RNA-immunoprecipitation (RBBP5/GCN5 interaction with LINC00930), siRNA, in vivo tumor model |
Journal of experimental & clinical cancer research |
Medium |
35209949
|
| 2019 |
PFKFB3 inhibition reduces TNF-α-induced endothelial proinflammatory responses (cytokines, ICAM-1, monocyte adhesion/transmigration) by suppressing IKKβ phosphorylation, IκBα phosphorylation/degradation, NF-κB-p65 nuclear translocation, and NF-κB DNA-binding activity. |
siRNA knockdown, PFKFB3 inhibitor, cytokine antibody array, monocyte adhesion/transmigration assay, NF-κB EMSA, immunofluorescence (p65 localization), western blotting |
Inflammation |
Medium |
30171427
|
| 2019 |
PFKFB3/HIF-1α form a positive feedback loop in hepatocellular carcinoma: HIF-1α drives PFKFB3 expression, and exogenous PFKFB3 overexpression in turn upregulates HIF-1α protein levels; HIF-1α deficiency impairs PFKFB3-induced sorafenib resistance. |
HIF-1α blockade, PFKFB3 overexpression, HIF-1α and PFKFB3 protein immunoblotting, apoptosis assay, GEO dataset validation |
Biochemical and biophysical research communications |
Low |
30981500
|
| 2023 |
cGAS-STING-IRF3 signaling pathway promotes renal fibrosis by upregulating PFKFB3 expression under hypoxia; inhibition of STING or IRF3 reverses elevated PFKFB3, placing PFKFB3 downstream of cGAS-STING-IRF3 in hypoxia-induced fibrosis. |
STING and IRF3 pharmacological inhibition, hypoxia-stimulated HK-2 cells, IRI mouse model, PFKFB3 expression immunoblotting, fibrosis markers |
Free radical biology & medicine |
Medium |
37714438
|
| 2022 |
HIF1A directly induces PFKFB3 expression in alveolar epithelial cells; alveolar epithelium-specific Pfkfb3 deletion dramatically increases acute lung injury severity, and pyruvate (restoring metabolic substrate) rescues both Pfkfb3loxP/loxP and Hif1aloxP/loxP mice from ALI. |
Alveolar epithelium-specific Pfkfb3-KO and Hif1a-KO mice (SPC-ER-Cre), injurious ventilation and acid instillation ALI models, intratracheal pyruvate rescue, human lung biopsy immunostaining |
JCI insight |
High |
36326834
|
| 2021 |
PFKFB3 inhibition in small cell lung carcinoma attenuates invasion/migration by downregulating YAP/TAZ signaling while increasing pLATS1 via activation of pMST1 and NF2, linking PFKFB3-driven glycolysis to regulation of the Hippo pathway. |
PFK158 inhibitor, shRNA stable knockdown, Hippo pathway component immunoblotting (pMST1, NF2, pLATS1, YAP/TAZ), xenograft model |
Oncogene |
Medium |
35804016
|
| 2019 |
RANKL-induced osteoclastogenesis increases PFKFB3 expression and glycolysis; L-lactate (the glycolytic product) partially reverses the suppression of osteoclastogenesis caused by PFKFB3 inhibition and abrogates the inhibitory effect on NF-κB and MAPK pathways, establishing lactate as a downstream mediator of PFKFB3's pro-osteoclast function. |
siRNA, PFKFB3 inhibitor (PFK15), L-lactate supplementation rescue, NF-κB and MAPK immunoblotting, lactate/glucose measurement, ovariectomy bone loss mouse model |
Journal of cellular and molecular medicine |
Medium |
31880389
|
| 2023 |
PFKFB3-derived lactate in renal fibroblasts promotes fibrotic activation; the glycolytic metabolite lactate directly promotes fibrotic phenotype in NRK-49F cells, and myofibroblast-specific PFKFB3 KO mice show substantially reduced fibrosis after UUO or IRI. |
Myofibroblast-specific Pfkfb3-KO mice (Pfkfb3f/f/PostnMCM), exogenous lactate supplementation, TGF-β1 stimulation, α-SMA and fibronectin markers, scRNA-seq reanalysis |
Cells |
Medium |
37626891
|
| 2024 |
SphK1/S1P signaling stabilizes PFKFB3 in endothelial cells to supply glycolytic energy for tumor angiogenesis; pharmacological SphK1 inhibition induces proteasomal degradation of PFKFB3, which can be reversed by S1P supplementation in an S1P receptor-dependent manner. |
SphK1 inhibitor (PF-543), Sphk1-KO mice, S1P supplementation rescue, PFKFB3 protein stability assay (proteasome inhibitor), DEN-induced primary HCC model, lentiviral SphK1 KD |
Journal of translational medicine |
Medium |
38200582
|
| 2023 |
In myeloid cells, PFKFB3-driven glycolysis stabilizes HIF1α, which alters macrophage phenotype to contribute to renal fibrosis; myeloid-specific PFKFB3-KO reduces M1 and M2 macrophage infiltration, suppresses macrophage-to-myofibroblast transition, and decreases kidney fibrosis. |
Myeloid-specific Pfkfb3-KO mice (Pfkfb3ΔMφ), UUO model, HIF1α stabilization assay, macrophage phenotyping by flow cytometry, scRNA-seq reanalysis |
Frontiers in immunology |
Medium |
38035106
|
| 2020 |
FAT10 upregulates PFKFB3 in osteosarcoma by directly binding to EGFR and inhibiting EGFR ubiquitination and degradation, thereby stabilizing EGFR which then promotes PFKFB3 expression and glycolysis. |
Co-immunoprecipitation (FAT10-EGFR), ubiquitination assay, siRNA knockdown, PFKFB3 expression immunoblot, glycolysis measurement, osteosarcoma tissue correlation |
American journal of cancer research |
Medium |
32775001
|
| 2023 |
PFKFB3-driven glycolysis in vascular smooth muscle cells promotes calcification through FoxO3 expression and lactate production; silencing PFKFB3 reduces FoxO3, and pyruvate/lactate supplementation reverses PFKFB3-depletion effects on ALP activity and OPG expression, establishing lactate as an osteogenic mediator downstream of PFKFB3. |
RNA-seq after PFKFB3 KD, FoxO3 silencing epistasis, lactate/pyruvate supplementation rescue, miR-26a/b-5p overexpression, in vivo vitamin D3 calcification model, VSMC osteogenic transdifferentiation assays |
FASEB journal |
Medium |
37682013
|
| 2022 |
PFKFB3 overexpression in bone marrow endothelial progenitor cells facilitates pro-apoptotic transcription factor FOXO3A and its downstream genes (p21, p27, FAS), activates NF-κB and E-selectin expression, and reduces SDF-1, impairing hematopoiesis-supporting function after chemotherapy; FOXO3A silencing rescues these effects. |
PFKFB3 overexpression/knockdown in BM EPCs, FOXO3A silencing rescue experiment, NF-κB activation immunoblot, E-selectin and SDF-1 measurement, BM EC-specific PFKFB3 overexpression transgenic mice, 5-FU chemotherapy model, patient BM EPC analysis |
Haematologica |
Medium |
35354250
|