title: Neuronal markers are expressed in human gliomas and NSE knockdown sensitizes glioblastoma cells to radiotherapy and temozolomide.
abstract: BACKGROUND
  Expression of neuronal elements has been identified in various glial tumors, and glioblastomas (GBMs) with neuronal differentiation patterns have reportedly been associated with longer survival. However, the neuronal class III β-tubulin has been linked to increasing malignancy in astrocytomas. Thus, the significance of neuronal markers in gliomas is not established.
  
  
  METHODS
  The expressions of class III β-tubulin, neurofilament protein (NFP), microtubule-associated protein 2 (MAP2) and neuron-specific enolase (NSE) were investigated in five GBM cell lines and two GBM biopsies with immunocytochemistry and Western blot. Moreover, the expression levels were quantified by real-time qPCR under different culture conditions. Following NSE siRNA treatment we used Electric cell-substrate impedance sensing (ECIS) to monitor cell growth and migration and MTS assays to study viability after irradiation and temozolomide treatment. Finally, we quantitated NSE expression in a series of human glioma biopsies with immunohistochemistry using a morphometry software, and collected survival data for the corresponding patients. The biopsies were then grouped according to expression in two halves which were compared by survival analysis.
  
  
  RESULTS
  Immunocytochemistry and Western blotting showed that all markers except NFP were expressed both in GBM cell lines and biopsies. Notably, qPCR demonstrated that NSE was upregulated in cellular stress conditions, such as serum-starvation and hypoxia, while we found no uniform pattern for the other markers. NSE knockdown reduced the migration of glioma cells, sensitized them to hypoxia, radio- and chemotherapy. Furthermore, we found that GBM patients in the group with the highest NSE expression lived significantly shorter than patients in the low-expression group.
  
  
  CONCLUSIONS
  Neuronal markers are aberrantly expressed in human GBMs, and NSE is consistently upregulated in different cellular stress conditions. Knockdown of NSE reduces the migration of GBM cells and sensitizes them to hypoxia, radiotherapy and chemotherapy. In addition, GBM patients with high NSE expression had significantly shorter survival than patients with low NSE expression. Collectively, these data suggest a role for NSE in the adaption to cellular stress, such as during treatment.
authors.affiliation: Department of Neurosurgery, Qilu Hospital, Shandong University, Jinan, PR China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yan
authors.forename: Kai Ove
authors.initials: KO
authors.lastname: Skaftnesmo
authors.forename: Lina
authors.initials: L
authors.lastname: Leiss
authors.forename: Linda
authors.initials: L
authors.lastname: Sleire
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Xingang
authors.initials: X
authors.lastname: Li
authors.forename: Per Øyvind
authors.initials: PØ
authors.lastname: Enger
mesh_terms: D000328:Adult; D000368:Aged; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001706:Biopsy; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000378:metabolism / Q000628:therapy; D015687:Cell Hypoxia / Q000187:drug effects / Q000528:radiation effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D003606:Dacarbazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D017097:Electric Impedance; D005260:Female; D055786:Gene Knockout Techniques; D005909:Glioblastoma / Q000378:metabolism / Q000628:therapy; D005910:Glioma / Q000378:metabolism / Q000628:therapy; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008875:Middle Aged; D016900:Neurofilament Proteins / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D000077204:Temozolomide; D014404:Tubulin / Q000378:metabolism; D055815:Young Adult
chemical_list: D018906:Antineoplastic Agents, Alkylating; D014408:Biomarkers, Tumor; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; D016900:Neurofilament Proteins; D014404:Tubulin; D003606:Dacarbazine; D010751:Phosphopyruvate Hydratase; D000077204:Temozolomide