The glycosylation characteristic of hepatoma-associated antigen HAb18G/CD147 in human hepatoma cells

Int J Biochem Cell Biol. 2006;38(11):1939-45. doi: 10.1016/j.biocel.2006.05.008. Epub 2006 May 27.

Abstract

HAb18G/CD147 is a highly glycosylated transmembrane protein belonging to the immunoglobulin superfamily. Our previous studies have demonstrated that overexpressing HAb18G/CD147 enhances the metastatic potentials of human hepatoma cells. In the present study, to investigate the glycosylation characteristic of HAb18G/CD147 in human hepatoma cells, HAb18G/CD147 was first purified from human FHCC-98 hepatoma cells by immunoaffinity chromatography, and then introduced into human fibroblasts culture system for matrix metalloproteinases induction. As a result, the elevated levels of matrix metalloproteinases secreted by fibroblasts were detected by gelatin zymography. The lysates of human hepatoma FHCC-98 cell revealed two major forms of HAb18G/CD147 (43-66 and 35 kDa) by western blot assay. To elucidate whether the variation of molecule size were caused by different glycosylation, two different approaches were employed to accomplish this goal: deglycosylation with N-glycosylation inhibitor tunicamycin or endoglycosidases. A single deglycosylated core protein with molecular weight approximately 27 kDa was obtained from both methods. Furthermore, the results of endoglycosidases treatment also showed that two forms of HAb18G/CD147 contain different types of oligosaccharide chains, thus sensitive to different endoglycosidase. In conclusion, the present study demonstrated that purified native HAb18G/CD147 has the bioactivity of stimulating human fibroblasts to produce elevated levels of matrix metalloproteinases, and that the two different forms of HAb18G/CD147 are derived from the single core protein but differ in their degree and types of glycosylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Neoplasm / chemistry
  • Antigens, Neoplasm / isolation & purification
  • Antigens, Neoplasm / metabolism
  • Basigin / chemistry
  • Basigin / isolation & purification
  • Basigin / metabolism*
  • Blotting, Western
  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology
  • Cell Line, Tumor
  • Electrophoresis, Polyacrylamide Gel
  • Glycoside Hydrolases / pharmacology
  • Glycosylation / drug effects
  • Humans
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • Matrix Metalloproteinases / metabolism
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / metabolism
  • Tunicamycin / pharmacology

Substances

  • Antigens, Neoplasm
  • Membrane Glycoproteins
  • Tunicamycin
  • Basigin
  • Glycoside Hydrolases
  • Matrix Metalloproteinases