Knock-down of Pdcd4 stimulates angiogenesis via up-regulation of angiopoietin-2

Biochim Biophys Acta. 2012 Apr;1823(4):789-99. doi: 10.1016/j.bbamcr.2012.01.006. Epub 2012 Jan 24.

Abstract

The tumor suppressor Pdcd4 is involved in multiple pathways. Considering its biological action conflicting data in the literature exist and, consequently, our own studies point to a cell type specific action of Pdcd4. In the present study, using several Pdcd4 knock down cell lines we succeeded to identify angiopoietin-2 (Ang-2) as a gene up-regulated on the mRNA and protein level. The subsequent enhanced peptide secretion forced wild type Bon-1 cells in a neoplastic direction demonstrated by increased proliferation and colony formation while cell adhesion was decreased. Most likely, the stimulation of Ang-2 is in part mediated by increased activation of AP-1 but different signal transduction pathways may also be involved since we found opposite activation of PI3K/Akt/mTOR and MAPK7ERK pathways (both known to regulate in Ang-2 expression). Ang-2 is a modulator of vascular remodeling. Therefore, we analyzed the effect of supernatants from Pdcd4 knock-down cell lines on endothelial cells. Again, we detected reduced cell adhesion and increased colony formation. Probably, the most impressive effect was described on tube formation in a model for angiogenesis. Tube length and junctions of endothelial cells treated with conditioned medium from Pdcd4 knock-down cells were considerably increased. Both, up-regulation of Ang-2 and down-regulation of Pdcd4 are described for many tumors. However, this is the first study showing a direct impact of Pdcd4 on Ang-2 levels and, thereby, angiogenesis. Our data suggest a completely new mechanism for Pdcd4 to act as a tumor suppressor rendering Pdcd4 an attractive target for new therapeutic strategies in cancer treatment.

MeSH terms

  • Angiopoietin-2 / genetics*
  • Angiopoietin-2 / metabolism
  • Apoptosis Regulatory Proteins / deficiency*
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism*
  • Cell Adhesion / drug effects
  • Cell Line
  • Cell Proliferation / drug effects
  • Colony-Forming Units Assay
  • Culture Media, Conditioned / pharmacology
  • Endothelial Cells / cytology
  • Endothelial Cells / drug effects
  • Endothelial Cells / metabolism
  • Gene Knockdown Techniques*
  • Humans
  • Neovascularization, Physiologic / drug effects
  • Neovascularization, Physiologic / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / metabolism
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Subcellular Fractions / drug effects
  • Subcellular Fractions / metabolism
  • Transfection
  • Up-Regulation / drug effects
  • Up-Regulation / genetics*

Substances

  • Angiopoietin-2
  • Apoptosis Regulatory Proteins
  • Culture Media, Conditioned
  • PDCD4 protein, human
  • RNA, Messenger
  • RNA, Small Interfering
  • RNA-Binding Proteins