E-selectin-dependent adhesion efficiency of colonic carcinoma cells is increased by genetic manipulation of their cell surface lysosomal membrane glycoprotein-1 expression levels

J Biol Chem. 1993 Jun 15;268(17):12675-81.

Abstract

Lysosomal membrane glycoprotein (lamp)-1 and lamp-2 are the most abundant glycoproteins within the lysosomal membrane. A small amount of lamp-1 and lamp-2 molecules, however, can be present on the cell surface. We have shown previously that highly metastatic colonic carcinoma L4 cells express more lamp-1 and lamp-2 on the cell surface than low metastatic SP cells (Saitoh, O., Wang, W.-L., Lotan, R., and Fukuda, M. (1992) J. Biol. Chem. 267, 5700-5711). Since lamp-1 and lamp-2 are the major carriers for poly-N-acetyllactosamines that are able to display sialyl-Le(x) termini, we sought to determine if an increased amount of lamp-1 on the cell surface would lead to increased expression of cell surface sialyl-Le(x) determinants and to the increased adhesion of those cells to E-selectin. Expression of increased amounts of lamp-1 on the cell surface was achieved either by overexpression of lamp-1 or by expressing a mutant lamp-1 molecule preferentially at the plasma membrane, rather than in lysosomes. Cells that express variable amounts of cell surface lamp-1 were tested for their adhesion to activated endothelial cells or E-selectin expressing Chinese hamster ovary cells. The results clearly show that the extent of adhesion to E-selectin and cell surface sialyl-Le(x) determinants is proportional to the amount of cell surface lamp-1. Moreover, it was demonstrated that such adhesion can be inhibited by soluble lamp-1 generated from Chinese hamster ovary cells expressing sialyl-Le(x) structures. These results indicate that lamp-1 can efficiently present ligands for E-selectin and at the same time can be a useful reagent for inhibition of E-selectin (and possibly P-selectin)-mediated interaction.

Publication types

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

MeSH terms

  • Animals
  • Antigens, CD*
  • Base Sequence
  • CHO Cells
  • Carbohydrate Conformation
  • Carbohydrate Sequence
  • Cell Adhesion Molecules / metabolism*
  • Cell Adhesion*
  • Colonic Neoplasms
  • Cricetinae
  • DNA
  • DNA Mutational Analysis
  • E-Selectin
  • Endothelium, Vascular / metabolism
  • Flow Cytometry
  • Gene Expression
  • Gene Library
  • Humans
  • Interleukin-1 / pharmacology
  • Lewis X Antigen / biosynthesis
  • Lewis X Antigen / metabolism*
  • Lysosomal Membrane Proteins
  • Membrane Glycoproteins / biosynthesis
  • Membrane Glycoproteins / metabolism*
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides
  • Tumor Cells, Cultured

Substances

  • Antigens, CD
  • Cell Adhesion Molecules
  • E-Selectin
  • Interleukin-1
  • Lewis X Antigen
  • Lysosomal Membrane Proteins
  • Membrane Glycoproteins
  • Oligodeoxyribonucleotides
  • DNA