ADAM15 overexpression in NIH3T3 cells enhances cell-cell interactions

Exp Cell Res. 2001 Nov 15;271(1):152-60. doi: 10.1006/excr.2001.5353.

Abstract

ADAM15 is a member of the family of metalloprotease-disintegrins that have been shown to interact with integrins in an RGD- and non-RGD-dependent manner. In the present study, we examined the effects of ADAM15 overexpression on cell-matrix and cell-cell interactions in NIH3T3 cells. Tetracycline-regulated ADAM15 overexpression in NIH3T3 cells leads to an inhibition of migration on a fibronectin-coated filter in a Boyden chamber assay and in a scratch wound model. The effects of ADAM15 overexpression on cell migration are not due to changes in matrix attachment or to the lack of extracellular signal-regulated kinase signaling response to PDGF or fibronectin. However, a decrease in monolayer permeability with ADAM15 overexpression and altered cell morphology suggest a possible increase in cell-cell interaction. Analysis of adhesion of NIH3T3 cells to a polyclonal population of cells retrovirally transduced to overexpress ADAM15 demonstrates a 45% increase in cell adhesion, compared with enhanced green fluorescent protein-expressing control cells. In addition, we demonstrate localization of HA-epitope-tagged ADAM15 to cell-cell contacts in an epithelial cell line that forms extensive cell-cell contact structures. Thus, overexpression of ADAM15 in NIH3T3 cells appears to enhance cell-cell interactions, as suggested by decreased cell migration, altered cell morphology at the wound edge, decreased monolayer permeability, and increased cell adhesion to monolayers of cells expressing ADAM15 by retroviral transduction.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • ADAM Proteins
  • Animals
  • Anticoagulants / pharmacology
  • Becaplermin
  • Cell Adhesion / physiology*
  • Cell Communication / physiology*
  • Cell Fractionation
  • Cell Line
  • Cell Movement / physiology*
  • Cell Size
  • Disintegrins / genetics
  • Disintegrins / metabolism*
  • Green Fluorescent Proteins
  • Humans
  • Indicators and Reagents / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism*
  • Mice
  • Mitogen-Activated Protein Kinases / metabolism
  • Platelet-Derived Growth Factor / pharmacology
  • Protein Synthesis Inhibitors / pharmacology
  • Proto-Oncogene Proteins c-sis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Retroviridae / genetics
  • Retroviridae / metabolism
  • Signal Transduction / physiology
  • Tetracycline / pharmacology
  • Transduction, Genetic

Substances

  • Anticoagulants
  • Disintegrins
  • Indicators and Reagents
  • Luminescent Proteins
  • Membrane Proteins
  • Platelet-Derived Growth Factor
  • Protein Synthesis Inhibitors
  • Proto-Oncogene Proteins c-sis
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Becaplermin
  • Mitogen-Activated Protein Kinases
  • ADAM Proteins
  • ADAM15 protein, human
  • Adam15 protein, mouse
  • Metalloendopeptidases
  • Tetracycline