title: NEK2 Promotes Aerobic Glycolysis in Multiple Myeloma Through Regulating Splicing of Pyruvate Kinase.
abstract: Aerobic glycolysis, a hallmark of cancer, is characterized by increased metabolism of glucose and production of lactate in normaxia. Recently, pyruvate kinase M2 (PKM2) has been identified as a key player for regulating aerobic glycolysis and promoting tumor cell proliferation and survival.
  
  
  
  Tandem affinity purification followed up by mass spectrometry (TAP-MS) and co-immunoprecipitation (Co-IP) were used to study the interaction between NIMA (never in mitosis gene A)-related kinase 2 (NEK2) and heterogeneous nuclear ribonucleoproteins (hnRNP) A1/2. RNA immunoprecipitation (RIP) was performed to identify NEK2 binding to PKM pre-mRNA sequence. Chromatin-immunoprecipitation (ChIP)-PCR was performed to analyze a transcriptional regulation of NEK2 by c-Myc. Western blot and real-time PCR were executed to analyze the regulation of PKM2 by NEK2.
  
  
  
  NEK2 regulates the alternative splicing of PKM immature RNA in multiple myeloma cells by interacting with hnRNPA1/2. RIP shows that NEK2 binds to the intronic sequence flanking exon 9 of PKM pre-mRNA. Knockdown of NEK2 decreases the ratio of PKM2/PKM1 and also other aerobic glycolysis genes including GLUT4, HK2, ENO1, LDHA, and MCT4. Myeloma patients with high expression of NEK2 and PKM2 have lower event-free survival and overall survival. Our data indicate that NEK2 is transcriptionally regulated by c-Myc in myeloma cells. Ectopic expression of NEK2 partially rescues growth inhibition and cell death induced by silenced c-Myc.
  
  
  
  Our studies demonstrate that NEK2 promotes aerobic glycolysis through regulating splicing of PKM and increasing the PKM2/PKM1 ratio in myeloma cells which contributes to its oncogenic activity.
authors.affiliation: Department of Medicine, Division of Hematology, Oncology and Blood and Marrow Transplantation and Holden Comprehensive Cancer Center, University of Iowa, 585 Newton Rd, 52242, Iowa City, IA, USA.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Gu
authors.affiliation: Department of Medicine, Division of Hematology, Oncology and Blood and Marrow Transplantation and Holden Comprehensive Cancer Center, University of Iowa, 585 Newton Rd, 52242, Iowa City, IA, USA.|Institute of Cancer Research, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
authors.forename: Jiliang
authors.initials: J
authors.lastname: Xia
authors.affiliation: Department of Medicine, Division of Hematology, Oncology and Blood and Marrow Transplantation and Holden Comprehensive Cancer Center, University of Iowa, 585 Newton Rd, 52242, Iowa City, IA, USA.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Medicine, Division of Hematology, Oncology and Blood and Marrow Transplantation and Holden Comprehensive Cancer Center, University of Iowa, 585 Newton Rd, 52242, Iowa City, IA, USA.
authors.forename: Ivana
authors.initials: I
authors.lastname: Frech
authors.affiliation: Department of Medicine, Division of Hematology, Oncology and Blood and Marrow Transplantation and Holden Comprehensive Cancer Center, University of Iowa, 585 Newton Rd, 52242, Iowa City, IA, USA.
authors.forename: Guido
authors.initials: G
authors.lastname: Tricot
authors.affiliation: Department of Medicine, Division of Hematology, Oncology and Blood and Marrow Transplantation and Holden Comprehensive Cancer Center, University of Iowa, 585 Newton Rd, 52242, Iowa City, IA, USA. fenghuang-zhan@uiowa.edu.
authors.forename: Fenghuang
authors.initials: F
authors.lastname: Zhan
mesh_terms: D000332:Aerobiosis; D002352:Carrier Proteins; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006019:Glycolysis; D006801:Humans; D008565:Membrane Proteins; D009101:Multiple Myeloma / Q000201:enzymology / Q000378:metabolism*; D000072379:NIMA-Related Kinases / Q000502:physiology*; D016271:Proto-Oncogene Proteins c-myc / Q000502:physiology; D011770:Pyruvate Kinase / Q000378:metabolism*; D012326:RNA Splicing; D013963:Thyroid Hormones; D000097810:Thyroid Hormone-Binding Proteins
chemical_list: D002352:Carrier Proteins; D008565:Membrane Proteins; D000072379:NIMA-Related Kinases; D016271:Proto-Oncogene Proteins c-myc; D011770:Pyruvate Kinase; D013963:Thyroid Hormones; D000097810:Thyroid Hormone-Binding Proteins; C089441:NEK2 protein, human
keywords: Alternative splicing; Multiple myeloma; NEK2; Pyruvate kinase