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RNA-seq analysis of PROX1 overexpression and knockdown in glioblastoma cell lines

ABSTRACT: RNA-seq analysis of PROX1 overexpression and knockdown in glioblastoma cell lines

PROVIDER: PRJEB27945 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR2715100.fastq.gz	Fastqsanger.gz
	ERR2715110.fastq.gz	Fastqsanger.gz
	ERR2715091.fastq.gz	Fastqsanger.gz
	ERR2715101.fastq.gz	Fastqsanger.gz
	ERR2715092.fastq.gz	Fastqsanger.gz

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RNA-seq analysis of PROX1 overexpression and knockdown in glioblastoma cell lines

Project description:Glioblastoma represents the most common and aggressive primary brain tumor type in adults. Stem cell regulatory pathways have been shown to be activated in gliomas supporting self-renewal, tumor maintenance and survival under stress. Glioblastoma stem-like phenotype, cell motility and tumor cell heterogeneity are considered significant hurdles to overcome for developing new treatment against these tumors. Transcription factor PROX1 has been associated with stem-like-phenotypes. Here, we overexpressed and suppressed PROX1 in glioma cell lines in order understand the gene expression regulated by this transcription factor.

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Gene expression profiles of lymphatic endothelial cells after knockdown of Prox1 and/or NR2F2

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Project description:Homeobox genes are a family of transcription factors that play a pivotal role in embryogenesis. Altered expression of Prospero-related homeobox gene 1 (PROX1) was recently associated with cancer, including oral squamous cell carcinoma (OSCC). This study aimed to investigate the functional role of PROX1 in oral carcinogenesis. PROX1 mRNA and protein expression levels were first investigated in 40 samples of OSCC and non-tumour margins. Methylation and amplification analysis was also performed to assess the epigenetic mechanisms involved in controlling PROX1 expression. OSCC cell line SCC9 was also transfected to stably express the PROX1 gene. Next, SCC9-PROX1-overexpressing cells and controls were then subjected to proliferation, differentiation, apoptosis, and microarray assays in vitro. OSCC samples had reduced PROX1 expression levels compared to non-tumour margins. PROX1 amplification was associated with better overall survival. PROX1 overexpression reduces cell proliferation by down-regulating cyclin D1 and up-regulating p21. PROX1-overexpressing cells also exhibited significantly reduced K18 and K19 expression.

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Prox1 inhibits axon outgrowth during central nervous system development

Project description:During central nervous system (CNS) development, proper and timely induction of axon elongation is critical for generating functional, mature neurons and neuronal networks. Despite the wealth of information on the action of extracellular cues, little is known about the intrinsic gene regulatory factors that control this developmental decision. Here we report the identification of Prox1, a homeobox transcription factor, as a key player in inhibiting axon elongation. Although Prox1 promotes acquisition of early neuronal identity and is expressed in nascent post-mitotic neurons, it is heavily down-regulated in the majority of terminally differentiated neurons, indicating a regulatory role in delaying axon outgrowth in newly formed neurons. Consistently, we show that Prox1 is sufficient to inhibit neurite extension in neuroblastoma cell lines. Furthermore, shRNA-mediated knock-down of Prox1 in Neuro2A cells induces the extension of neurites. More importantly, Prox1 overexpression suppresses axon elongation in primary neuronal cultures as well as in the developing mouse brain, while Prox1 knock-down promotes axon outgrowth. Mechanistically, RNA-Seq analysis reveals that Prox1 affects critical pathways for neuronal maturation and neurite extension. Interestingly, Prox1 strongly inhibits many components of Ca2+ signaling pathway, an important mediator of axon extension and neuronal maturation. In accordance, Prox1 represses Ca2+ entry upon KCl-mediated depolarization and reduce CREB phosphorylation. These observations suggest that Prox1 acts as a potent suppressor of axon elongation by inhibiting Ca2+ signaling pathway. This action may provide the appropriate time window for nascent neurons to find the correct position in the CNS prior to initiation of axon elongation.

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