title: Differential analysis of glioblastoma multiforme proteome by a 2D-DIGE approach.
abstract: BACKGROUND
  Genomics, transcriptomics and proteomics of glioblastoma multiforme (GBM) have recently emerged as possible tools to discover therapeutic targets and biomarkers for new therapies including immunotherapy. It is well known that macroscopically complete surgical excision, radiotherapy and chemotherapy have therapeutic limitations to improve survival in these patients. In this study, we used a differential proteomic-based technique (2D-Difference Gel Electrophoresis) coupled with matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry to identify proteins that may serve as brain tumor antigens in new therapeutic assays. Five samples of patients presenting a GBM and five samples of microscopically normal brain tissues derived from brain epileptic surgery specimen were labeled and run in 2D-PAGE (Two-Dimensional Polyacrylamide Gel Electrophoresis) with an internal pool sample on each gel. Five gels were matched and compared with DIA (Difference In-gel Analysis) software. Differential spots were picked, in-gel digested and peptide mass fingerprints were obtained.
  
  
  RESULTS
  From 51 protein-spots significantly up-regulated in GBM samples, mass spectrometry (MS) identified twenty-two proteins. The differential expression of a selected protein set was first validated by western-blotting, then tested on large cohorts of GBM specimens and non-tumor tissues, using immunohistochemistry and real-time RT-PCR.
  
  
  CONCLUSIONS
  Our results confirmed the importance of previously described proteins in glioma pathology and their potential usefulness as biological markers but also revealed some new interesting targets for future therapies.
authors.affiliation: Centre Régional de Lutte contre le Cancer; Rennes, France. b.collet@rennes.fnclcc.fr.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Collet
authors.forename: Nathalie
authors.initials: N
authors.lastname: Guitton
authors.forename: Stephan
authors.initials: S
authors.lastname: Saïkali
authors.forename: Tony
authors.initials: T
authors.lastname: Avril
authors.forename: Charles
authors.initials: C
authors.lastname: Pineau
authors.forename: Abderrahmane
authors.initials: A
authors.lastname: Hamlat
authors.forename: Jean
authors.initials: J
authors.lastname: Mosser
authors.forename: Véronique
authors.initials: V
authors.lastname: Quillien