Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

MYCN overexpression in SH-SY5Y/6TR(EU)/pTrex-Dest-30/MYCN cell line

ABSTRACT: MYCN overexpression in the doxycline MYCN inducible cell line SH-SY5Y/6TR(EU)/pTrex-Dest-30/MYCN (SY5Y-MYCN) using the dynamic transcriptome analysis (DTA) protocol. Samples at different time-points after MYCN over-expression and uninduced controls (0h) were sequenced in duplicates. The experimental setup consists of [A] total mRNA at 0h, 1h, 4h, 24h, which corresponds to the standard mRNA-seq protocol, [B] 4-thioUridine (4sU) labelled mRNA 30 min before extraction at time-points 0h, 1h, 4h, which is the freshly transcribed mRNA within the last 30 min before the time-point and [C] the counter-part, 4sU unlabelled mRNA 30 min before extraction which corresponds to the mRNA from before 30 min of extraction. The samples were sequenced on an Illumina GA IIx using the Illumina protocols.

INSTRUMENT(S): Illumina Genome Analyzer IIx

ORGANISM(S): Homo sapiens

SUBMITTER: Thomas Schwarzl

PROVIDER: E-MTAB-2691 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Integrative omics reveals MYCN as a global suppressor of cellular signalling and enables network-based therapeutic target discovery in neuroblastoma.

Duffy David J DJ Krstic Aleksandar A Halasz Melinda M Schwarzl Thomas T Fey Dirk D Iljin Kristiina K Mehta Jai Prakash JP Killick Kate K Whilde Jenny J Turriziani Benedetta B Haapa-Paananen Saija S Fey Vidal V Fischer Matthias M Westermann Frank F Henrich Kai-Oliver KO Bannert Steffen S Higgins Desmond G DG Kolch Walter W

Oncotarget 20151201 41

Despite intensive study, many mysteries remain about the MYCN oncogene's functions. Here we focus on MYCN's role in neuroblastoma, the most common extracranial childhood cancer. MYCN gene amplification occurs in 20% of cases, but other recurrent somatic mutations are rare. This scarcity of tractable targets has hampered efforts to develop new therapeutic options. We employed a multi-level omics approach to examine MYCN functioning and identify novel therapeutic targets for this largely un-drugga ...[more]

PMID: 26673823

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Project description:MYCN is an oncogene amplified in approximately 20% of all neuroblastomas and 50% of high risk neuroblastoma. It is a transcription factor regulating the expression of genes involved in proliferation and apoptosis.We would like to identify genes potentially regulated by MYCN in a conditional MYCN expression system. The SHEP Tet21N system is a conditional, tetracycline-regulated MYCN expression system established in the MYCN non-amplified SHEP neuroblastoma cell line (Lutz et al., 1996). Expression of MYCN is switched off by the addition of tetracycline to the growth medium. Levels of MYCN protein and RNA in the absence of tetracycline are comparable with those present in MYCN amplified cell lines (Bell et al., 2006). By performing microarray analyses on the Tet21n cell lines in the presence and absence of MYCN we hope to identify genes whose expression had been altered by MYCN either upregulated or downregulated and may be potential MYCN targets. The altered expression of some of these genes will then be validated using quantitative RTPCR and a selection will be further investigated to determine if they are MYCN transcriptional targets and may be potential candidates for anti-MYCN therapies for neuroblastoma Specific objectives 1) To investigate genes which are altered by 2 fold or more in the presence of or absence of MYCN in Tet21N cells. Refs: Bell, E., Premkumar, R., Carr, J., Lu, X., Lovat, P.E., Kees, U.R., Lunec, J. & Tweddle, D.A. (2006). The role of MYCN in the failure of MYCN amplified neuroblastoma cell lines to G1 arrest after DNA damage. Cell Cycle, 5, 2639-47. Lutz, W., Stohr, M., Schurmann, J., Wenzel, A., Lohr, A. & Schwab, M. (1996). Conditional expression of N-myc in human neuroblastoma cells increases expression of alpha-prothymosin and ornithine decarboxylase and accelerates progression into S-phase early after mitogenic stimulation of quiescent cells. Oncogene, 13, 803-12.

Dataset Information

MYCN overexpression in SH-SY5Y/6TR(EU)/pTrex-Dest-30/MYCN cell line

Publications

Integrative omics reveals MYCN as a global suppressor of cellular signalling and enables network-based therapeutic target discovery in neuroblastoma.

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