ADAM-10-mediated N-cadherin cleavage is protein kinase C-alpha dependent and promotes glioblastoma cell migration

J Neurosci. 2009 Apr 8;29(14):4605-15. doi: 10.1523/JNEUROSCI.5126-08.2009.

Abstract

MMPs (matrix metalloproteinases) and the related "a disintegrin and metalloproteinases" (ADAMs) promote tumorigenesis by cleaving extracellular matrix and protein substrates, including N-cadherin. Although N-cadherin is thought to regulate cell adhesion, migration, and invasion, its role has not been characterized in glioblastomas (GBMs). In this study, we investigated the expression and function of posttranslational N-cadherin cleavage in GBM cells as well as its regulation by protein kinase C (PKC). N-Cadherin cleavage occurred at a higher level in glioblastoma cells than in non-neoplastic astrocytes. Treatment with the PKC activator phorbol 12-myristate 13-acetate (PMA) increased N-cadherin cleavage, which was reduced by pharmacological inhibitors and short interfering RNA (siRNA) specific for ADAM-10 or PKC-alpha. Furthermore, treatment of GBM cells with PMA induced the translocation of ADAM-10 to the cell membrane, the site at which N-cadherin was cleaved, and this translocation was significantly reduced by the PKC-alpha inhibitor Gö6976 [12-(2-cyanoethyl)-6,7,12,13-tetrahydro-13-methyl-5-oxo-5H-indolo[2,3-a]pyrrolo[3,4-c]carbazole] or PKC-alpha short hairpin RNA. In functional studies, N-cadherin cleavage was required for GBM cell migration, as depletion of N-cadherin cleavage by N-cadherin siRNA, ADAM-10 siRNA, or a cleavage-site mutant N-cadherin, decreased GBM cell migration. Together, these results suggest that N-cadherin cleavage is regulated by a PKC-alpha-ADAM-10 cascade in GBM cells and may be involved in mediating GBM cell migration.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • ADAM Proteins / chemistry
  • ADAM Proteins / physiology*
  • ADAM10 Protein
  • Amyloid Precursor Protein Secretases / chemistry
  • Amyloid Precursor Protein Secretases / physiology*
  • Antigens, CD / chemistry
  • Antigens, CD / metabolism*
  • Cadherins / chemistry
  • Cadherins / metabolism*
  • Cell Line, Tumor
  • Cell Migration Inhibition / genetics
  • Cell Migration Inhibition / physiology
  • Cell Movement / genetics
  • Cell Movement / physiology*
  • Cells, Cultured
  • Glioblastoma / enzymology*
  • Glioblastoma / pathology
  • Humans
  • Hydrolysis
  • Membrane Proteins / chemistry
  • Membrane Proteins / physiology*
  • Mutation
  • Protein Kinase C-alpha / physiology*

Substances

  • Antigens, CD
  • CDH2 protein, human
  • Cadherins
  • Membrane Proteins
  • Protein Kinase C-alpha
  • Amyloid Precursor Protein Secretases
  • ADAM Proteins
  • ADAM10 Protein
  • ADAM10 protein, human