Abstract
MicroRNAs function as negative regulators of posttranscriptional gene expression, having major roles in cellular differentiation. Several neuroblastoma cell lines can be induced to undergo differentiation by all-trans-retinoic acid (ATRA) and are used for modeling signaling pathways involved in this process. To identify miRNAs contributing to differentiation, we profiled 364 loci following ATRA treatment of neuroblastoma cell lines and found miR-10a and miR-10b to be highly overexpressed in SK-N-BE, LAN5 and SHSY-5Y. Ectopic overexpression of these miRNAs led to a major reprogramming of the transcriptome and a differentiated phenotype that was similar to that induced by ATRA in each of these cell lines. One of the predicted downregulated miR-10a/b targets was nuclear receptor corepressor 2 (NCOR2), a corepressor of gene transcription, which is known to suppress neurite outgrowth. NCOR2 was experimentally validated as a direct target of miR-10a/b, and siRNA-mediated inhibition of this mRNA alone resulted in neural cell differentiation. Moreover, induction of differentiation could be blocked by ectopic upregulation of NCOR2 using an expression construct lacking the miR-10a/b 3′ untranslated region target site. We conclude that miR-10a/b has major roles in the process of neural cell differentiation through direct targeting of NCOR2, which in turn induces a cascade of primary and secondary transcriptional alterations, including the downregulation of MYCN.
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Abbreviations
- ATRA:
-
all-trans-retinoic acid
- NCOR2:
-
nuclear receptor co-repressor 2
- MYCN:
-
v-myc myelocytomatosis viral related oncogene
- HOX:
-
homeobox gene
- TUBB3, tubulin:
-
beta 3
- GAP43:
-
growth associated protein 43
- miRNA:
-
microRNA
- miR:
-
microRNA
- UTR:
-
untranslated regions
- GO:
-
gene ontology
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Acknowledgements
This work was supported in part by Science Foundation Ireland (07/IN.1/B1776), Children's Medical and Research Foundation, Cancer Research Ireland and the US National Institutes of Health (5R01CA127496). TB was supported by a Marie Curie Cofund EU Fellowship (CEMP; jointly funded by the National Biophotonics and Imaging Platform, Higher Education Authority, Ireland).
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Foley, N., Bray, I., Watters, K. et al. MicroRNAs 10a and 10b are potent inducers of neuroblastoma cell differentiation through targeting of nuclear receptor corepressor 2. Cell Death Differ 18, 1089–1098 (2011). https://doi.org/10.1038/cdd.2010.172
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DOI: https://doi.org/10.1038/cdd.2010.172
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