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David Ruff Life Technologies, United States of America |
Abstract
MicroRNAs (miRNAs) bind to mRNA and influence gene expression by destabilizing the target mRNA or repressing translation. miRNAs have a key regulatory role in cellular processes such as apoptosis, oncogenesis, differentiation and development. miRNAs have important functions in neuronal differentiation and recent reports detail essential roles of several miRNA species. The NTERA2 pluripotent human embryonic carcinoma cell line is a simple in vitro model system for characterizing molecular events occurring in early neuronal differentiation. Exposure of NTERA2 cells to retinoic acid induces cellular differentiation into the neuroectodermal lineage. We harvested cell cultures for miRNA and protein expression studies throughout a 14-day differentiation time course study. miRNA signatures of targets were obtained using the MegaPlexTM Preamp Primer Pools and TaqMan® Array MicroRNA cards. In parallel, protein expression profiles were generated using TaqMan® Protein Assays. Our data indicate dynamic regulation of miRNA species throughout the time course – many miRNA species are upregulated during the differentiation process. Several upregulated miRNA species such as miR-21, miR-125b and miR-145 have been reported to interact with specific mRNAs and modulate their respective protein levels. For instance, a major player in the biology of cellular response, the p53 protein, is directly impacted by miRNA cues. We quantified a panel of proteins influenced by miRNA regulation during the temporal differentiation of NTERA2 cells. Our experimental approach and qPCR assay tools have great value for researchers studying the interplay between miRNA dynamics and protein output.
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