Downregulation of erythropoietin receptor by overexpression of phospholipase C-gamma 1 is critical for decrease on focal adhesion in transformed cells

Cell Oncol (Dordr). 2011 Feb;34(1):11-21. doi: 10.1007/s13402-010-0001-9. Epub 2011 Jan 18.

Abstract

Background: Phospholipase C-γl (PLC-γl) is known to play a critical role in cell adhesion and migration and is highly expressed in metastatic tumors. In the current study, we found that cells transformed by PLC overexpression (PLC-γl cells) exhibited a marked decrease in expression of the Epo receptor (EpoR). Here, we assessed the role of EpoR-dependent signaling pathways in PLC-γl-dependent regulation of cell adhesion and migration.

Methods: Expression and phosphorylation of EpoR and its functional role in PLC-γl cells were evaluated by immunoblot analysis or cell adhesion assay. The mechanism for PLC-γ1-induced EpoR downregulation was analyzed by blockage of proteosomal degradation with MG132. EpoR expression was also confirmed in colorectal cancer tissues in which PLC-γl was highly expressed.

Results: EpoR was present on rat fibroblasts, where it functionally active and capable of increasing cell adhesion and migratory activity. However, PLC-γl cells significantly decreased the Epo-dependent effects via ubiquitination-proteosomal degradation of EpoR. A marked decrease of EpoR expression was confirmed in colorectal cancer tissues that showed high-level of PLC-γl expression.

Conclusion: The Epo/EpoR complex plays a critical role in the adhesion and migration of rat fibroblasts, and its functional inactivation is associated with PLC-γl-dependent reduction of cell-matrix adhesion and this also affects cell migration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Adhesion / drug effects
  • Cell Movement / drug effects
  • Cell Transformation, Neoplastic / drug effects
  • Cell Transformation, Neoplastic / metabolism*
  • Cell Transformation, Neoplastic / pathology
  • Colorectal Neoplasms / enzymology
  • Colorectal Neoplasms / pathology
  • Down-Regulation* / drug effects
  • Erythropoietin / pharmacology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Focal Adhesions / drug effects
  • Focal Adhesions / metabolism*
  • Humans
  • Male
  • Mice
  • Middle Aged
  • PC12 Cells
  • Paxillin / metabolism
  • Phospholipase C gamma / metabolism*
  • Proteasome Endopeptidase Complex / metabolism
  • Protein Processing, Post-Translational / drug effects
  • Rats
  • Receptors, Erythropoietin / genetics*
  • Receptors, Erythropoietin / metabolism
  • Signal Transduction / drug effects

Substances

  • Paxillin
  • Receptors, Erythropoietin
  • Erythropoietin
  • Phospholipase C gamma
  • Proteasome Endopeptidase Complex