title: JAK2/STAT3 targeted therapy suppresses tumor invasion via disruption of the EGFRvIII/JAK2/STAT3 axis and associated focal adhesion in EGFRvIII-expressing glioblastoma.
abstract: BACKGROUND
  As a commonly mutated form of the epidermal growth factor receptor, EGFRvIII strongly promotes glioblastoma (GBM) tumor invasion and progression, but the mechanisms underlying this promotion are not fully understood.
  
  
  METHODS
  Through gene manipulation, we established EGFRvIII-, wild-type EGFR-, and vector-expressing GBM cells. We used cDNA microarrays, bioinformatics analysis, target-blocking migration and invasion assays, Western blotting, and an orthotopic U87MG GBM model to examine the phenotypic shifts and treatment effects of EGFRvIII expression in vitro and in vivo. Confocal imaging, co-immunoprecipitation, and siRNA assays detected the focal adhesion-associated complex and their relationships to the EGFRvIII/JAK2/STAT3 axis in GBM cells.
  
  
  RESULTS
  The activation of JAK2/STAT3 signaling is vital for promoting migration and invasion in EGFRvIII-GBM cells. AG490 or WP1066, the JAK2/STAT3 inhibitors, specifically destroyed EGFRvIII/JAK2/STAT3-related focal adhesions and depleted the activation of EGFR/Akt/FAK and JAK2/STAT3 signaling, thereby abolishing the ability of EGFRvIII-expressing GBM cells to migrate and invade. Furthermore, the RNAi silencing of JAK2 in EGFRvIII-expressing GBM cells significantly attenuated their ability to migrate and invade; however, as a result of a potential EGFRvIII-JAK2-STAT3 activation loop, neither EGFR nor STAT3 knockdown yielded the same effects. Moreover, AG490 or JAK2 gene knockdown greatly suppressed tumor invasion and progression in the U87MG-EGFRvIII orthotopic models.
  
  
  CONCLUSIONS
  Taken together, our data demonstrate that JAK2/STAT3 signaling is essential for EGFRvIII-driven migration and invasion by promoting focal adhesion and stabilizing the EGFRvIII/JAK2/STAT3 axis. Targeting JAK2/STAT3 therapy, such as AG490, may have potential clinical implications for the tailored treatment of GBM patients bearing EGFRvIII-positive tumors.
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Qifan
authors.initials: Q
authors.lastname: Zheng
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Lei
authors.initials: L
authors.lastname: Han
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Yucui
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Jing
authors.initials: J
authors.lastname: Tian
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Zhaoyu
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Xiuzhi
authors.initials: X
authors.lastname: Jia
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Tao
authors.initials: T
authors.lastname: Jiang
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Jianning
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Kang
authors.affiliation: Department of Immunology, Harbin Medical University; Heilongjiang Provincial Key Laboratory for Infection and Immunity, Harbin, China (Q.-F.Z., Y.-C.D., J.T., W.H., Z.-Y.L., X.-Z.J., H.R.); Department of Neurosurgery, Beijing Tiantan Hospital, Capital Medical University, Beijing, China (T.J.); College of Bioinformatics, Harbin Medical University, Harbin, China (X.L.); Department of Neurosurgery, Tianjin Medical University General Hospital; Laboratory of Neuro-Oncology, Tianjin Neurological Institute; Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education; Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin, China (L.H., C.-S.K., J.-N.Z.); Chinese Glioma Cooperative Group (CGCG) (L.H., T.J., C.-S.K.).
authors.forename: Huan
authors.initials: H
authors.lastname: Ren
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage*; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000008:administration & dosage; D066246:ErbB Receptors / Q000378:metabolism*; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism; D005909:Glioblastoma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D053614:Janus Kinase 2 / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D050796:STAT3 Transcription Factor / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D020032:Tyrphostins / Q000008:administration & dosage*
chemical_list: D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D020032:Tyrphostins; C095512:alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide; C410723:epidermal growth factor receptor VIII; D066246:ErbB Receptors; D053614:Janus Kinase 2
keywords: EGFRvIII; JAK2/STAT3; focal adhesion; glioblastoma; invasion