Identification of targets of miR-200b by a SILAC-based quantitative proteomic approach

• 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.

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