Over-expression of beta-1,4-galactosyltransferase V increases the growth of astrocytoma cell line

J Exp Clin Cancer Res. 2002 Sep;21(3):409-14.

Abstract

Our previous study showed that the gene expression of beta-1,4-galactosyltransferase V (beta-1,4-GalT V), preferentially galactosylating GlcNAc1-->6Man of oligosaccharides, increased in the process of astrocytoma progress, with the highest level in grade IV astrocytoma. To investigate the function of this beta-1,4-GalT in cell proliferation, the sense and antisense cDNA of beta-1,4-GalT V was constructed as pcDNA3-HA-GalT V and pcDNA3-anti-GalT V respectively and transfected into SHG cell, a kind of human astrocytoma cell line. The transfection was confirmed with Northern and Western blot assay. It was found that the growth of SHG/HA-GalT V in serum-containing medium was faster than that of mock-transfectant with the vector pcDNA3, whereas the growth of SHG/GalTV-AS was slower than that of mock-transfectant. GalTV-HA/SHG showed a stronger capability for colony formation than that of GalTV-AS/SHG as evaluated by anchorage-independent growth in soft agar assay. This result was consistent with that of the growth curve. By RCA-1 lectin assay, the galactosylation on the surface of GalTV-HA/SHG and SHG/GalTV-AS was stained stronger (P<0.001) and weaker (P<0.05) respectively compared with the mock transfectant. This indicates that beta-1,4-GalT V was involved in the malignant phenotype of astrocytoma cells, possibly causing the high galactosylation on the cell surface.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Astrocytoma / enzymology*
  • Astrocytoma / pathology
  • Blotting, Northern
  • Blotting, Western
  • Brain Neoplasms / enzymology*
  • Brain Neoplasms / pathology
  • Cell Division
  • Colony-Forming Units Assay
  • Galactose / metabolism
  • Galactosyltransferases / metabolism*
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Lectins / metabolism
  • Plasmids
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Lectins
  • Galactosyltransferases
  • beta-1,4-galactosyltransferase V
  • Galactose