title: Shift of microRNA profile upon glioma cell migration using patient-derived spheroids and serum-free conditions.
abstract: Glioblastoma multiforme (GBM) is the most frequent malignant primary brain tumor. A major reason for the overall median survival being only 14.6 months is migrating tumor cells left behind after surgery. Another major reason is tumor cells having a so-called cancer stem cell phenotype being therefore resistant towards traditional chemo- and radiotherapy. A group of novel molecular targets are microRNAs (miRNAs). MiRNAs are small non-coding RNAs exerting post-transcriptional regulation of gene expression. The aim of this study was to identify differentially expressed miRNAs in migrating GBM cells using serum-free stem cell conditions. We used patient-derived GBM spheroid cultures for a novel serum-free migration assay. MiRNA expression of migrating tumor cells isolated at maximum migration speed was compared with corresponding spheroids using an OpenArray Real-Time PCR System. The miRNA profiling revealed 30 miRNAs to be differentially expressed. In total 13 miRNAs were upregulated and 17 downregulated in migrating cells compared to corresponding spheroids. The three most deregulated miRNAs, miR-1227 (up-regulated), miR-32 (down-regulated) and miR-222 (down-regulated), were experimentally overexpressed. A non-significantly increased migration rate was observed after miR-1227 overexpression. A significantly reduced migration rate was observed after miR-32 and miR-222 overexpression. In conclusion a shift in microRNA profile upon glioma cell migration was identified using an assay avoiding serum-induced migration. Both the miRNA profiling and the functional validation suggested that miR-1227 may be associated with increased migration and miR-32 and miR-222 with decreased migration. These miRNAs may represent potential novel targets in migrating glioma cells.
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 3rd floor, 5000, Odense C, Denmark.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19, 5000, Odense C, Denmark.|Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Sune
authors.initials: S
authors.lastname: Munthe
authors.affiliation: Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19, 5000, Odense C, Denmark.|Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Bo
authors.initials: B
authors.lastname: Halle
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 3rd floor, 5000, Odense C, Denmark.
authors.forename: Henning B
authors.initials: HB
authors.lastname: Boldt
authors.affiliation: Department of Nuclear Medicine, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Helle
authors.initials: H
authors.lastname: Christiansen
authors.affiliation: Faculty of Health Sciences, Lundbeckfonden Center of Excellence NanoCAN and Molecular Oncology, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 25, 5000, Odense C, Denmark.
authors.forename: Steffen
authors.initials: S
authors.lastname: Schmidt
authors.affiliation: Regulus Therapeutics, San Diego, CA, USA.
authors.forename: Vivek
authors.initials: V
authors.lastname: Kaimal
authors.affiliation: Regulus Therapeutics, San Diego, CA, USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Xu
authors.affiliation: Regulus Therapeutics, San Diego, CA, USA.
authors.forename: Sonya
authors.initials: S
authors.lastname: Zabludoff
authors.affiliation: Faculty of Health Sciences, Lundbeckfonden Center of Excellence NanoCAN and Molecular Oncology, Institute of Molecular Medicine, University of Southern Denmark, Winsløwparken 25, 5000, Odense C, Denmark.
authors.forename: Jan
authors.initials: J
authors.lastname: Mollenhauer
authors.affiliation: Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19, 5000, Odense C, Denmark.|Department of Neurosurgery, Odense University Hospital, Sdr. Boulevard 29, 5000, Odense C, Denmark.
authors.forename: Frantz R
authors.initials: FR
authors.lastname: Poulsen
authors.affiliation: Department of Pathology, Odense University Hospital, Winsløwparken 15, 3rd floor, 5000, Odense C, Denmark. bjarne.winther.kristensen@rsyd.dk.|Institute of Clinical Research, University of Southern Denmark, Winsløwparken 19, 5000, Odense C, Denmark. bjarne.winther.kristensen@rsyd.dk.
authors.forename: Bjarne W
authors.initials: BW
authors.lastname: Kristensen
mesh_terms: D001932:Brain Neoplasms / Q000378:metabolism*; D002465:Cell Movement; D016895:Culture Media, Serum-Free; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D018874:Spheroids, Cellular; D014407:Tumor Cells, Cultured
chemical_list: D016895:Culture Media, Serum-Free; D035683:MicroRNAs
keywords: Glioblastoma; MicroRNA; Migration; Serum-free; Target