Identification of targets of miR-200b by a SILAC-based quantitative proteomic approach
Publication Type:Journal Article
Source:EuPA Open Proteomics, Elsevier, Volume 4, p.10-17 (2014)
Keywords:amino acid analysis, analytic method, article, bioinformatics, calpastatin, cancer growth, Cell culture, cell cycle progression, cell proliferation, cell protein, cellular distribution, controlled study, cyclin dependent kinase 6, down regulation, exportin 2, fibronectin, fibronectin 1, fibrosarcoma cell line, galectin 3, Gene Expression Regulation, gene function, gene overexpression, high mobility group B1 protein, histidine nucleotide binding protein 1, human, human cell, initiation factor 5A, Isotope Labeling, lipocortin 1, liquid chromatography, microRNA 200b, nuclear protein, nucleotide binding protein, p21 activated kinase 2, peroxiredoxin 1, polymerase chain reaction, programmed cell death protein 5, protein determination, protein expression, protein function, protein localization, protein RNA binding, protein targeting, proteomics, quantitative study, RNA analysis, stable isotope labeling by aminoacids in cell culture, tandem mass spectrometry, tumor necrosis factor receptor associated factor 1, unclassified drug, validation study
miRNAs regulate gene expression by binding to cognate mRNAs causing mRNA degradation or translational repression. Mass spectrometry-based proteomic analysis is being widely used to identify miRNA targets. The miR-200b miRNA cluster is often overexpressed in multiple cancer types, but the identity of the targets remains elusive. Using SILAC-based analysis, we examined the effects of overexpression of a miR-200b mimic or a control miRNA in fibrosarcoma cells. We identified around 300 potential targets of miR-200b based on a change in the expression of protein levels. We validated a subset of potential targets at the transcript level using quantitative PCR.
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