A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene

Neuroblastoma is the most common extracranial solid tumor of infancy. Amplification of MYCN oncogene is found in approximately 20 % of all neuroblastoma patients and correlates with advanced disease stages, rapid tumor progression, and poor prognosis, making this gene an obvious therapeutic target....

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Main Authors:	Sidarovich, Viktoryia, Adami, Valentina, Quattrone, Alessandro
Format:	Online
Language:	English
Published:	Springer US 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4067544/

id	pubmed-4067544
recordtype	oai_dc
spelling	pubmed-40675442014-07-02 A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene Sidarovich, Viktoryia Adami, Valentina Quattrone, Alessandro Research Neuroblastoma is the most common extracranial solid tumor of infancy. Amplification of MYCN oncogene is found in approximately 20 % of all neuroblastoma patients and correlates with advanced disease stages, rapid tumor progression, and poor prognosis, making this gene an obvious therapeutic target. However, being a transcriptional factor MYCN is difficult for pharmacological targeting, and there are currently no clinical trials aiming MYCN protein directly. Here we describe an alternative approach to address deregulated MYCN expression. In particular, we focus on the role of a 3′ untranslated region (3′UTR) of the MYCN gene in the modulation of its mRNA fate and identification of compounds able to affect it. The luciferase reporter construct with the full length MYCN 3′UTR was generated and subsequently integrated in the CHP134 neuroblastoma cell line. After validation, the assay was used to screen a 2000 compound library. Molecules affecting luciferase activity were checked for reproducibility and counter-screened for promoter effects and cytotoxic activity resulting in selection of four hits. We propose this cell-based reporter gene assay as a valuable tool to screen chemical libraries for compounds modulating post-transcriptional control mechanisms. Identification of such compounds could potentially result in development of clinically relevant therapeutics for various diseases including neuroblastoma. Springer US 2014-02-11 2014 /pmc/articles/PMC4067544/ /pubmed/24515800 http://dx.doi.org/10.1007/s12033-014-9739-z Text en © The Author(s) 2014 Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Sidarovich, Viktoryia Adami, Valentina Quattrone, Alessandro
spellingShingle	Sidarovich, Viktoryia Adami, Valentina Quattrone, Alessandro A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene
author_facet	Sidarovich, Viktoryia Adami, Valentina Quattrone, Alessandro
author_sort	Sidarovich, Viktoryia
title	A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene
title_short	A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene
title_full	A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene
title_fullStr	A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene
title_full_unstemmed	A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene
title_sort	cell-based high-throughput screen addressing 3′utr-dependent regulation of the mycn gene
description	Neuroblastoma is the most common extracranial solid tumor of infancy. Amplification of MYCN oncogene is found in approximately 20 % of all neuroblastoma patients and correlates with advanced disease stages, rapid tumor progression, and poor prognosis, making this gene an obvious therapeutic target. However, being a transcriptional factor MYCN is difficult for pharmacological targeting, and there are currently no clinical trials aiming MYCN protein directly. Here we describe an alternative approach to address deregulated MYCN expression. In particular, we focus on the role of a 3′ untranslated region (3′UTR) of the MYCN gene in the modulation of its mRNA fate and identification of compounds able to affect it. The luciferase reporter construct with the full length MYCN 3′UTR was generated and subsequently integrated in the CHP134 neuroblastoma cell line. After validation, the assay was used to screen a 2000 compound library. Molecules affecting luciferase activity were checked for reproducibility and counter-screened for promoter effects and cytotoxic activity resulting in selection of four hits. We propose this cell-based reporter gene assay as a valuable tool to screen chemical libraries for compounds modulating post-transcriptional control mechanisms. Identification of such compounds could potentially result in development of clinically relevant therapeutics for various diseases including neuroblastoma.
publisher	Springer US
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4067544/
_version_	1612104985214976000

A Cell-Based High-Throughput Screen Addressing 3′UTR-Dependent Regulation of the MYCN Gene

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