Effect of microRNA-206 on cytoskeleton remodelling by downregulating Cdc42 in MDA-MB-231 cells

Tumori. 2010 Sep-Oct;96(5):751-5. doi: 10.1177/030089161009600518.

Abstract

Aims and background: MicroRNAs are small, noncoding, single-stranded RNAs that regulate gene expression post-transcriptionally. miR-206 is known to play an important role in breast cancer metastasis. When we sought to predict the target of miR-206 by Targetscan, Pictar and miRanda, we found Cdc42 was a potential one. In this study, we transfected miR-206 into MDA-MB-231 cells and examined Cdc42 protein expression as well as MMP-2 and MMP-9, which are also associated with metastasis of breast cancer. Since Cdc42 is involved in filopodia and invadopodia formation, we examined the morphological changes of MDA-MB-231 cells.

Methods and study design: miR-206 mimics were transfected into MDA-MB-231 cells using LipofectamineTM 2000. Protein expression was detected by Western blot. Cells were stained with FITC-phalloidin to visualize F-actin. Invasive ability and migratory ability were examined by invasion assay and migration assay in vitro.

Results: Cdc42, MMP-2 and MMP-9 were downregulated on the protein level. The formation of filopodia, which requires Cdc42, was inhibited in miR-206 transfected cells, even under the stimulation of EGF. The invasion and migration of MDA-MB-231 cells in vitro was inhibited by miR-206 too.

Conclusions: The results suggest that miR-206 may suppress invasion and migration of MDA-MB-231 cells in vitro partly via regulating actin cytoskeleton remodelling such as filopodia formation.

MeSH terms

  • Actins / metabolism
  • Blotting, Western
  • Cell Line, Tumor
  • Cell Movement
  • Cytoskeleton / genetics
  • Cytoskeleton / metabolism*
  • Down-Regulation
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • MicroRNAs / metabolism*
  • Microscopy, Fluorescence
  • Polymerase Chain Reaction
  • Transfection
  • cdc42 GTP-Binding Protein / metabolism*

Substances

  • Actins
  • MIRN206 microRNA, human
  • MicroRNAs
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9
  • cdc42 GTP-Binding Protein