ADAM-10-mediated N-cadherin cleavage is protein kinase C-alpha dependent and promotes glioblastoma cell migration.
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.
Kohutek
Zachary A
ZA
Departments of Pathology and Molecular Physiology and Biological Physics, University of Virginia Health System, Charlottesville, VA 22908, USA. zsk3k@virginia.edu
diPierro
Charles G
CG
Redpath
Gerard T
GT
Hussaini
Isa M
IM
D051722:ADAM Proteins / Q000737:chemistry / Q000502:physiology*; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000737:chemistry / Q000502:physiology*; D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism*; D015820:Cadherins / Q000737:chemistry / Q000378:metabolism*; D045744:Cell Line, Tumor; D002464:Cell Migration Inhibition / Q000235:genetics / Q000502:physiology; D002465:Cell Movement / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D005909:Glioblastoma / Q000201:enzymology* / Q000473:pathology; D006801:Humans; D006868:Hydrolysis; D008565:Membrane Proteins / Q000737:chemistry / Q000502:physiology*; D009154:Mutation; D051571:Protein Kinase C-alpha / Q000502:physiology*
D015703:Antigens, CD; C050361:CDH2 protein, human; D015820:Cadherins; D008565:Membrane Proteins; D051571:Protein Kinase C-alpha; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human