Involvement of cell surface glycans in adhesion of human colon carcinoma cells to liver tissue in a frozen section assay: role of endo-beta-galactosidase-sensitive structures.

Abstract

Adhesion of human colon carcinoma variant cell lines expressing different levels of the cell surface sialyl Lewis X (sLeX) antigen to frozen sections of mouse liver was examined. KM12-HX cells that bound the monoclonal antibody (mAb) FH6 (anti-sLeX) and thus expressed a high level of sLeX demonstrated a greater degree of adhesion to liver sections than their low-binding counterparts, KM12-LX cells. The adhesion of KM12-HX cells to liver sections was partially blocked by mAb FH6, but not by another anti-sLeX mAb, KM93. The adhesion was Ca2+ dependent but was not inhibited by anti-E-selectin. Endo-beta-galactosidase treatment significantly reduced adhesion and resulted in the loss of cell surface binding sites for mAb FH6. O-linked oligosaccharides from KM12-HX cells incubated in the presence of p-nitrophenyl-N-acetylgalactosaminide were fractionated by a combination of gel filtration, anion exchange chromatography, and normal phase high-performance liquid chromatography. The structure of a mAb FH6-reactive and endo-beta-galactosidase-sensitive glycan was estimated by matrix-assisted laser desorption ionization time-of-flight mass spectrometry in a post source decay mode and by glycosidase digestions to be NeuAc alpha2-3Gal beta1-4GlcNAc beta1-3Gal beta1-4Glc-NAc beta1-3Gal beta1-4(+/-Fuc alpha1-3)GlcNAc beta1-6(NeuAc alpha2-3Gal beta1-3)GalNAc-pNP. Mild detergent lysates of mouse liver surface-labeled with sulfo-NHS biotin were incubated with glutaraldehyde-fixed monolayers of KM12-HX cells, and bound components were isolated after EDTA treatment. A Mr 49,000 component that bound only to KM12-HX cells and not to KM12-LX cells was identified.

• Authors: M Ota, N Takamura, T Irimura
• Journal: Cancer research (Cancer Res) Vol. 60 Issue 18 Pg. 5261-8 (Sep 15 2000) ISSN: 0008-5472 [Print] United States
• PMID: 11016656 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Oligosaccharides
• Proteins
• Sialyl Lewis X Antigen
• Glycoside Hydrolases
• keratan-sulfate endo-1,4-beta-galactosidase
• beta-Galactosidase

Topics

• Animals
• Antibodies, Monoclonal (immunology, metabolism)
• CHO Cells
• Carbohydrate Sequence
• Cell Adhesion (physiology)
• Chromatography, Affinity
• Colonic Neoplasms (immunology, metabolism, pathology)
• Cricetinae
• Flow Cytometry
• Frozen Sections
• Glycoside Hydrolases
• Humans
• Liver (cytology, immunology, metabolism)
• Male
• Mice
• Mice, Inbred BALB C
• Molecular Sequence Data
• Oligosaccharides (biosynthesis, immunology, metabolism)
• Proteins (immunology, metabolism)
• Sialyl Lewis X Antigen
• Tumor Cells, Cultured
• beta-Galactosidase (metabolism, pharmacology)