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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Apoptosis / genetics
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Astrocytes / metabolism
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Astrocytes / pathology
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Biomarkers, Tumor / metabolism
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Brain Neoplasms / genetics*
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Brain Neoplasms / pathology*
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Brain Neoplasms / surgery
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Cell Differentiation / genetics*
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Cell Line, Tumor
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Female
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Gene Expression Profiling
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Gene Expression Regulation, Neoplastic
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Glial Fibrillary Acidic Protein / metabolism
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Glioblastoma / genetics*
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Glioblastoma / pathology*
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Glioblastoma / surgery
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Humans
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Membrane Proteins / metabolism
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Mice
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Mice, Inbred BALB C
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MicroRNAs / genetics
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MicroRNAs / metabolism*
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Myeloid Cell Leukemia Sequence 1 Protein / metabolism
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Neoplastic Stem Cells / metabolism*
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Neoplastic Stem Cells / pathology
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Nestin / metabolism
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Neurons / metabolism
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Neurons / pathology
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Oligonucleotide Array Sequence Analysis
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Phosphoproteins / metabolism
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Spheroids, Cellular / metabolism
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Spheroids, Cellular / pathology
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Tubulin / metabolism
Substances
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Biomarkers, Tumor
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Glial Fibrillary Acidic Protein
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MCL1 protein, human
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Membrane Proteins
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MicroRNAs
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Myeloid Cell Leukemia Sequence 1 Protein
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Nestin
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Phosphoproteins
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SPRY1 protein, human
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TUBB3 protein, human
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Tubulin
Grants and funding
This work was supported by Grants from the Instituto de Salud Carlos III- FIS, Spanish Ministries of Science, Innovation and Health (PI11/00684 to R.M., PI10/02002 to J.L.F.-L. and PI12/0202 to J.A.M.-C.); from Red Temática de Investigación Cooperativa en Cáncer (RTICC, RD12/0036/0063 to J.A.M.-C. and RD06/0020/0074 and RD12/0036/0022 to J.L.F.-L.); from Fundación Mutua Madrileña (FMM 8553/2011 to R.M.); from the Spanish Association Against Cancer (AECC) Scientific Foundation (to R.M.); and from Instituto de Formación e Investigación Marques de Valdecilla (API2011-04). B.A. work was supported by a predoctoral fellowship from Instituto de Salud Carlos III- FIS.The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.