Abstract
Glioblastoma (GBM) is the most common primary brain tumor, with poor prognosis and a lack of effective therapeutic options. The aberrant expression of transcription factor REST (repressor element 1-silencing transcription factor) had been reported in different kinds of tumors. However, the function of REST and its mechanisms in GBM remain elusive. Here, REST expression was inhibited by siRNA silencing in U-87 and U-251 GBM cells. Then CCK-8 assay showed significantly decreased cell proliferation, and the inhibition of migration was verified by scratch wound healing assay and transwell assay. Using cell cycle analysis and Annexin V/PI straining assay, G1 phase cell cycle arrest was found to be a reason for the suppression of cell proliferation and migration upon REST silencing, while apoptosis was not affected by REST silencing. Further, the detection of REST-downstream genes involved in cytostasis and migration inhibition demonstrated that CCND1 and CCNE1 were reduced; CDK5R1, BBC3, EGR1, SLC25A4, PDCD7, MAPK11, MAPK12, FADD and DAXX were enhanced, among which BBC3 and DAXX were direct targets of REST, as verified by ChIP (chromatin immunoprecipitation) and Western blotting. These data suggested that REST is a master regulator that maintains GBM cells proliferation and migration, partly through regulating cell cycle by repressing downstream genes, which might represent a potential target for GBM therapy.
Keywords:
NRSF; REST; glioblastoma; migration; proliferation.
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Apoptosis Regulatory Proteins / genetics
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Apoptosis Regulatory Proteins / metabolism
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Cell Line, Tumor
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Cell Movement*
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Cell Proliferation*
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Co-Repressor Proteins
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Cyclin D1 / genetics
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Cyclin D1 / metabolism
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Cyclin E / genetics
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Cyclin E / metabolism
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Early Growth Response Protein 1 / genetics
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Early Growth Response Protein 1 / metabolism
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Fas-Associated Death Domain Protein / genetics
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Fas-Associated Death Domain Protein / metabolism
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G1 Phase
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Glioblastoma / metabolism*
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Humans
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Membrane Transport Proteins
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Mitogen-Activated Protein Kinases / genetics
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Mitogen-Activated Protein Kinases / metabolism
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Molecular Chaperones
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Oncogene Proteins / genetics
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Oncogene Proteins / metabolism
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / metabolism
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Repressor Proteins / genetics
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Repressor Proteins / metabolism*
Substances
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Adaptor Proteins, Signal Transducing
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Apoptosis Regulatory Proteins
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BBC3 protein, human
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CCND1 protein, human
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CCNE1 protein, human
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Carrier Proteins
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Co-Repressor Proteins
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Cyclin E
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DAXX protein, human
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EGR1 protein, human
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Early Growth Response Protein 1
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FADD protein, human
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Fas-Associated Death Domain Protein
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Membrane Transport Proteins
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Molecular Chaperones
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Nerve Tissue Proteins
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Nuclear Proteins
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Oncogene Proteins
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Proto-Oncogene Proteins
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RE1-silencing transcription factor
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Repressor Proteins
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SLC15A4 protein, human
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neuronal Cdk5 activator (p25-p35)
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Cyclin D1
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Mitogen-Activated Protein Kinases