A Proteomics Approach to Investigate miR-153-3p and miR-205-5p Targets in Neuroblastoma Cells

PLoS One. 2015 Dec 3;10(12):e0143969. doi: 10.1371/journal.pone.0143969. eCollection 2015.

Abstract

MicroRNAs are key regulators associated with numerous diseases. In HEK293 cells, miR-153-3p and miR-205-5p down-regulate alpha-synuclein (SNCA) and Leucine-rich repeat kinase 2 (LRRK2), two key proteins involved in Parkinson's disease (PD). We have used two-dimensional gel electrophoresis (2D-PAGE) coupled to mass spectrometry (MS) to identify a spectrum of miR-153-3p and miR-205-5p targets in neuronal SH-SY5Y cells. We overexpressed and inhibited both microRNAs in SH-SY5Y cells and through comparative proteomics profiling we quantified ~240 protein spots from each analysis. Combined, thirty-three protein spots were identified showing significant (p-value < 0.05) changes in abundance. Modulation of miR-153-3p resulted in seven up-regulated proteins and eight down-regulated proteins. miR-205 modulation resulted in twelve up-regulated proteins and six down-regulated proteins. Several of the proteins are associated with neuronal processes, including peroxiredoxin-2 and -4, cofilin-1, prefoldin 2, alpha-enolase, human nucleoside diphosphate kinase B (Nm23) and 14-3-3 protein epsilon. Many of the differentially expressed proteins are involved in diverse pathways including metabolism, neurotrophin signaling, actin cytoskeletal regulation, HIF-1 signaling and the proteasome indicating that miR-153-3p and miR-205-5p are involved in the regulation of a wide variety of biological processes in neuroblastoma cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle
  • Cell Line, Tumor
  • Electrophoresis, Gel, Two-Dimensional
  • Gene Expression Regulation, Neoplastic / genetics
  • Humans
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Neoplasm Proteins / metabolism*
  • Neuroblastoma / genetics*
  • Neuroblastoma / metabolism
  • Neuroblastoma / pathology
  • Proteomics*
  • Transcription, Genetic / genetics

Substances

  • MIRN153 microRNA, human
  • MIRN205 microRNA, human
  • MicroRNAs
  • Neoplasm Proteins

Grants and funding

This work was funded by grants from The Norwegian Research Council, The Western Norway Health Authority and The Norwegian Center for Movement Disorders. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.