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Analysis of SOX2-Regulated Transcriptome in Glioma Stem Cells

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Glioblastoma is the most malignant brain tumor in adults and is associated with poor survival despite multimodal treatments. Glioma stem-like cells (GSCs) are cells functionally defined by their self-renewal potential and the ability to reconstitute the original tumor upon orthotopic implantation. They have been postulated to be the culprit of glioma chemo- and radio-resistance ultimately leading to relapse. Understanding the molecular circuits governing the GSC compartment is essential. SOX2, a critical transcription regulator of embryonic and neural stem cell function, is deregulated in GSCs however; the precise molecular pathways regulated by this gene in GSCs remain poorly understood.

Results: We performed a genome-wide analysis of SOX2-regulated transcripts in GSCs, using a microarray. We identified a total of 2048 differentially expressed coding transcripts and 261 non-coding transcripts. Cell adhesion and cell-cell signaling are among the most enriched terms using Gene Ontology (GO) classification. The pathways altered after SOX2 down-modulation includes multiple cellular processes such as amino-acid metabolism and intercellular signaling cascades. We also defined and classified the set of non-coding transcripts differentially expressed regulated by SOX2 in GSCs, and validated two of them.

Conclusions: We present a comprehensive analysis of the transcriptome controlled by SOX2 in GSCs, gaining insights in the understanding of the potential roles of SOX2 in glioblastoma.

No MeSH data available.


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Transcripts regulated by SOX2.(A) Schematic representation of the research design employed to uncover the SOX2 transcriptome in GSC11 cells. (B) qRT-PCR and western blot confirmation of SOX2 inhibition in GSC11 cells after 72h of si-SOX2 or si-Scramble (si-Scrbl) transfection. SOX2 relative mRNA levels are presented as 2-ΔΔCt standardized with their constitutive gene GAPDH. Each bar represents the mean ± SD. For western blot tubulin was used as housekeeping control and shown as a representative blot of four independent experiments.
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pone.0163155.g001: Transcripts regulated by SOX2.(A) Schematic representation of the research design employed to uncover the SOX2 transcriptome in GSC11 cells. (B) qRT-PCR and western blot confirmation of SOX2 inhibition in GSC11 cells after 72h of si-SOX2 or si-Scramble (si-Scrbl) transfection. SOX2 relative mRNA levels are presented as 2-ΔΔCt standardized with their constitutive gene GAPDH. Each bar represents the mean ± SD. For western blot tubulin was used as housekeeping control and shown as a representative blot of four independent experiments.

Mentions: Since SOX2 is a key driver in the maintenance of the GSCs phenotype and therefore in the perpetuation of this devastating tumor we down-regulated the expression of this gene in the GSC11 cell line in four independent experiments, using a SOX2 specific siRNA (Fig 1A). The efficiency of SOX2 knockdown was assessed by real-time PCR and western blot (Fig 1B) and was also confirmed in our array results. Microarray data identified a total of 2048 differentially expressed coding transcripts and 261 non-coding transcripts (B value >0) (Fig 1A and S1 Table).


Analysis of SOX2-Regulated Transcriptome in Glioma Stem Cells
Transcripts regulated by SOX2.(A) Schematic representation of the research design employed to uncover the SOX2 transcriptome in GSC11 cells. (B) qRT-PCR and western blot confirmation of SOX2 inhibition in GSC11 cells after 72h of si-SOX2 or si-Scramble (si-Scrbl) transfection. SOX2 relative mRNA levels are presented as 2-ΔΔCt standardized with their constitutive gene GAPDH. Each bar represents the mean ± SD. For western blot tubulin was used as housekeeping control and shown as a representative blot of four independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5036841&req=5

pone.0163155.g001: Transcripts regulated by SOX2.(A) Schematic representation of the research design employed to uncover the SOX2 transcriptome in GSC11 cells. (B) qRT-PCR and western blot confirmation of SOX2 inhibition in GSC11 cells after 72h of si-SOX2 or si-Scramble (si-Scrbl) transfection. SOX2 relative mRNA levels are presented as 2-ΔΔCt standardized with their constitutive gene GAPDH. Each bar represents the mean ± SD. For western blot tubulin was used as housekeeping control and shown as a representative blot of four independent experiments.
Mentions: Since SOX2 is a key driver in the maintenance of the GSCs phenotype and therefore in the perpetuation of this devastating tumor we down-regulated the expression of this gene in the GSC11 cell line in four independent experiments, using a SOX2 specific siRNA (Fig 1A). The efficiency of SOX2 knockdown was assessed by real-time PCR and western blot (Fig 1B) and was also confirmed in our array results. Microarray data identified a total of 2048 differentially expressed coding transcripts and 261 non-coding transcripts (B value >0) (Fig 1A and S1 Table).

View Article: PubMed Central - PubMed

ABSTRACT

Introduction: Glioblastoma is the most malignant brain tumor in adults and is associated with poor survival despite multimodal treatments. Glioma stem-like cells (GSCs) are cells functionally defined by their self-renewal potential and the ability to reconstitute the original tumor upon orthotopic implantation. They have been postulated to be the culprit of glioma chemo- and radio-resistance ultimately leading to relapse. Understanding the molecular circuits governing the GSC compartment is essential. SOX2, a critical transcription regulator of embryonic and neural stem cell function, is deregulated in GSCs however; the precise molecular pathways regulated by this gene in GSCs remain poorly understood.

Results: We performed a genome-wide analysis of SOX2-regulated transcripts in GSCs, using a microarray. We identified a total of 2048 differentially expressed coding transcripts and 261 non-coding transcripts. Cell adhesion and cell-cell signaling are among the most enriched terms using Gene Ontology (GO) classification. The pathways altered after SOX2 down-modulation includes multiple cellular processes such as amino-acid metabolism and intercellular signaling cascades. We also defined and classified the set of non-coding transcripts differentially expressed regulated by SOX2 in GSCs, and validated two of them.

Conclusions: We present a comprehensive analysis of the transcriptome controlled by SOX2 in GSCs, gaining insights in the understanding of the potential roles of SOX2 in glioblastoma.

No MeSH data available.


Related in: MedlinePlus