Abstract
•Demonstrates SILAC coupled robust mass spectrometry-based approach to identify protein targets regulated by miRNAs.•Reports identification and quantification of proteins dysregulated by miR-200b, a crucial miRNA known to be involved in epithelial to mesenchymal transition.•Reports the finding of potential novel targets of miR-200b.
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.