title: MicroRNA-566 activates EGFR signaling and its inhibition sensitizes glioblastoma cells to nimotuzumab.
abstract: BACKGROUND
  Epidermal growth factor receptor (EGFR) is amplified in 40% of human glioblastomas. However, most glioblastoma patients respond poorly to anti-EGFR therapy. MicroRNAs can function as either oncogenes or tumor suppressor genes, and have been shown to play an important role in cancer cell proliferation, invasion and apoptosis. Whether microRNAs can impact the therapeutic effects of EGFR inhibitors in glioblastoma is unknown.
  
  
  METHODS
  miR-566 expression levels were detected in glioma cell lines, using real-time quantitative RT-PCR (qRT-PCR). Luciferase reporter assays and Western blots were used to validate VHL as a direct target gene of miR-566. Cell proliferation, invasion, cell cycle distribution and apoptosis were also examined to confirm whether miR-566 inhibition could sensitize anti-EGFR therapy.
  
  
  RESULTS
  In this study, we demonstrated that miR-566 is up-regulated in human glioma cell lines and inhibition of miR-566 decreased the activity of the EGFR pathway. Lentiviral mediated inhibition of miR-566 in glioblastoma cell lines significantly inhibited cell proliferation and invasion and led to cell cycle arrest in the G0/G1 phase. In addition, we identified von Hippel-Lindau (VHL) as a novel functional target of miR-566. VHL regulates the formation of the β-catenin/hypoxia-inducible factors-1α complex under miR-566 regulation.
  
  
  CONCLUSIONS
  miR-566 activated EGFR signaling and its inhibition sensitized glioblastoma cells to anti-EGFR therapy.
authors.forename: Kai-Liang
authors.initials: KL
authors.lastname: Zhang
authors.forename: Xuan
authors.initials: X
authors.lastname: Zhou
authors.forename: Lei
authors.initials: L
authors.lastname: Han
authors.forename: Lu-Yue
authors.initials: LY
authors.lastname: Chen
authors.forename: Ling-Chao
authors.initials: LC
authors.lastname: Chen
authors.forename: Zhen-Dong
authors.initials: ZD
authors.lastname: Shi
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.forename: Yu
authors.initials: Y
authors.lastname: Ren
authors.forename: Jing-Xuan
authors.initials: JX
authors.lastname: Yang
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Frank
authors.forename: Chuan-Bao
authors.initials: CB
authors.lastname: Zhang
authors.forename: Jun-Xia
authors.initials: JX
authors.lastname: Zhang
authors.forename: Pei-Yu
authors.initials: PY
authors.lastname: Pu
authors.forename: Jian-Ning
authors.initials: JN
authors.lastname: Zhang
authors.forename: Tao
authors.initials: T
authors.lastname: Jiang
authors.affiliation: 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 300052, China. Eric.J.Wagner@uth.tmc.edu.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Wagner
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Chun-Sheng
authors.initials: CS
authors.lastname: Kang
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005909:Glioblastoma / Q000235:genetics* / Q000378:metabolism; D064593:Heterografts; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D014162:Transfection; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; C000591379:MIRN566 microRNA, human; D035683:MicroRNAs; C501466:nimotuzumab; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C512478:EGFR protein, human; D066246:ErbB Receptors; C497446:VHL protein, human