title: Involvement of miRNAs in the differentiation of human glioblastoma multiforme stem-like cells.
abstract: Glioblastoma multiforme (GBM)-initiating cells (GICs) represent a tumor subpopulation with neural stem cell-like properties that is responsible for the development, progression and therapeutic resistance of human GBM. We have recently shown that blockade of NFκB pathway promotes terminal differentiation and senescence of GICs both in vitro and in vivo, indicating that induction of differentiation may be a potential therapeutic strategy for GBM. MicroRNAs have been implicated in the pathogenesis of GBM, but a high-throughput analysis of their role in GIC differentiation has not been reported. We have established human GIC cell lines that can be efficiently differentiated into cells expressing astrocytic and neuronal lineage markers. Using this in vitro system, a microarray-based high-throughput analysis to determine global expression changes of microRNAs during differentiation of GICs was performed. A number of changes in the levels of microRNAs were detected in differentiating GICs, including over-expression of hsa-miR-21, hsa-miR-29a, hsa-miR-29b, hsa-miR-221 and hsa-miR-222, and down-regulation of hsa-miR-93 and hsa-miR-106a. Functional studies showed that miR-21 over-expression in GICs induced comparable cell differentiation features and targeted SPRY1 mRNA, which encodes for a negative regulator of neural stem-cell differentiation. In addition, miR-221 and miR-222 inhibition in differentiated cells restored the expression of stem cell markers while reducing differentiation markers. Finally, miR-29a and miR-29b targeted MCL1 mRNA in GICs and increased apoptosis. Our study uncovers the microRNA dynamic expression changes occurring during differentiation of GICs, and identifies miR-21 and miR-221/222 as key regulators of this process.
authors.affiliation: Division of Oncology, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain ; Human Oncology and Pathogenesis Program, Memorial Sloan-Kettering Cancer Center, New York, New York, United States of America.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Aldaz
authors.forename: Ainara
authors.initials: A
authors.lastname: Sagardoy
authors.forename: Lorena
authors.initials: L
authors.lastname: Nogueira
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Guruceaga
authors.forename: Lara
authors.initials: L
authors.lastname: Grande
authors.forename: Jason T
authors.initials: JT
authors.lastname: Huse
authors.forename: Maria A
authors.initials: MA
authors.lastname: Aznar
authors.forename: Ricardo
authors.initials: R
authors.lastname: Díez-Valle
authors.forename: Sonia
authors.initials: S
authors.lastname: Tejada-Solís
authors.forename: Marta M
authors.initials: MM
authors.lastname: Alonso
authors.forename: Jose L
authors.initials: JL
authors.lastname: Fernandez-Luna
authors.forename: Jose A
authors.initials: JA
authors.lastname: Martinez-Climent
authors.forename: Raquel
authors.initials: R
authors.lastname: Malumbres
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001253:Astrocytes / Q000378:metabolism / Q000473:pathology; D014408:Biomarkers, Tumor / Q000378:metabolism; D001932:Brain Neoplasms / Q000235:genetics* / Q000473:pathology* / Q000601:surgery; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005909:Glioblastoma / Q000235:genetics* / Q000473:pathology* / Q000601:surgery; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000378:metabolism* / Q000473:pathology; D064231:Nestin / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D010750:Phosphoproteins / Q000378:metabolism; D018874:Spheroids, Cellular / Q000378:metabolism / Q000473:pathology; D014404:Tubulin / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D005904:Glial Fibrillary Acidic Protein; C579095:MCL1 protein, human; D008565:Membrane Proteins; D035683:MicroRNAs; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D064231:Nestin; D010750:Phosphoproteins; C486055:SPRY1 protein, human; C498585:TUBB3 protein, human; D014404:Tubulin