RESEARCH A Cell-Based High-Throughput Screen Addressing 30UTR-Dependent Regulation of the MYCN Gene

The Author(s) 2014. This article is published with open access at Springerlink.com Abstract Neuroblastoma is the most common extracra-nial solid tumor of infancy. Amplification of MYCN oncogene is found in approximately 20 % of all neuro-blastoma 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 phar-macological 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 30 untranslated region (30UTR) of the MYCN gene in the modulation of its mRNA fate and identification of com-pounds able to affect it. The luciferase reporter construct with the full length MYCN 30UTR 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 selec-tion 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 mech-anisms. Identification of such compounds could potentially result in development of clinically relevant therapeutics for various diseases including neuroblastoma