title: Transmembrane protein CD9 is glioblastoma biomarker, relevant for maintenance of glioblastoma stem cells.
abstract: The cancer stem cell model suggests that glioblastomas contain a subpopulation of stem-like tumor cells that reproduce themselves to sustain tumor growth. Targeting these cells thus represents a novel treatment strategy and therefore more specific markers that characterize glioblastoma stem cells need to be identified. In the present study, we performed transcriptomic analysis of glioblastoma tissues compared to normal brain tissues revealing sensible up-regulation of CD9 gene. CD9 encodes the transmembrane protein tetraspanin which is involved in tumor cell invasion, apoptosis and resistance to chemotherapy. Using the public REMBRANDT database for brain tumors, we confirmed the prognostic value of CD9, whereby a more than two fold up-regulation correlates with shorter patient survival. We validated CD9 gene and protein expression showing selective up-regulation in glioblastoma stem cells isolated from primary biopsies and in primary organotypic glioblastoma spheroids as well as in U87-MG and U373 glioblastoma cell lines. In contrast, no or low CD9 gene expression was observed in normal human astrocytes, normal brain tissue and neural stem cells. CD9 silencing in three CD133+ glioblastoma cell lines (NCH644, NCH421k and NCH660h) led to decreased cell proliferation, survival, invasion, and self-renewal ability, and altered expression of the stem-cell markers CD133, nestin and SOX2. Moreover, CD9-silenced glioblastoma stem cells showed altered activation patterns of the Akt, MapK and Stat3 signaling transducers. Orthotopic xenotransplantation of CD9-silenced glioblastoma stem cells into nude rats promoted prolonged survival. Therefore, CD9 should be further evaluated as a target for glioblastoma treatment.
authors.affiliation: Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, Slovenia.
authors.forename: Neža
authors.initials: N
authors.lastname: Podergajs
authors.affiliation: Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, Slovenia.
authors.forename: Helena
authors.initials: H
authors.lastname: Motaln
authors.affiliation: Department of Biochemistry, Blood Transfusion Centre of Slovenia, 1000 Ljubljana, Slovenia.
authors.forename: Uroš
authors.initials: U
authors.lastname: Rajčević
authors.affiliation: Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, Slovenia.
authors.forename: Urška
authors.initials: U
authors.lastname: Verbovšek
authors.affiliation: Department of Neurosurgery, University Medical Centre, University of Ljubljana, 1000 Ljubljana, Slovenia.
authors.forename: Marjan
authors.initials: M
authors.lastname: Koršič
authors.affiliation: Department of Biomedicine, University of Bergen, 5009 Bergen, Norway.
authors.forename: Nina
authors.initials: N
authors.lastname: Obad
authors.affiliation: Department of Biomedicine, University of Bergen, 5009 Bergen, Norway.
authors.forename: Heidi
authors.initials: H
authors.lastname: Espedal
authors.affiliation: Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, Slovenia.
authors.forename: Miloš
authors.initials: M
authors.lastname: Vittori
authors.affiliation: Division of Neurosurgical Research, Department of Neurosurgery, University of Heidelberg, 69120 Heidelberg, Germany.
authors.forename: Christel
authors.initials: C
authors.lastname: Herold-Mende
authors.affiliation: Department of Biomedicine, University of Bergen, 5009 Bergen, Norway.
authors.forename: Hrvoje
authors.initials: H
authors.lastname: Miletic
authors.affiliation: Department of Biomedicine, University of Bergen, 5009 Bergen, Norway.|NorLux Neuro-Oncology Laboratory, Centre de Recherche Public de la Santé, 1526 Luxembourg, Luxembourg.
authors.forename: Rolf
authors.initials: R
authors.lastname: Bjerkvig
authors.affiliation: Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, 1000 Ljubljana, Slovenia.|Department of Biochemistry, Faculty of Chemistry and Chemical Engineering, University of Ljubljana, 1000 Ljubljana, Slovenia.
authors.forename: Tamara Lah
authors.initials: TL
authors.lastname: Turnšek
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic; D005909:Glioblastoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D009924:Organ Culture Techniques; D034622:RNA Interference; D011923:Rats, Nude; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016019:Survival Analysis; D060245:Tetraspanin 29 / Q000235:genetics* / Q000378:metabolism; D014183:Transplantation, Heterologous; D015854:Up-Regulation
chemical_list: D014408:Biomarkers, Tumor; D060245:Tetraspanin 29
keywords: CD9; biomarker; glioblastoma stem cells; neural stem cells; tetraspanin