Human colon cancer cell lines permanently expressing alpha 2,6-sialylated sugar chains by transfection with rat beta-galactoside alpha 2,6 sialyltransferase cDNA

Biochem Biophys Res Commun. 1995 Jun 15;211(2):554-61. doi: 10.1006/bbrc.1995.1849.

Abstract

A role of alpha 2,6-linked sialic acid in the development of an invasive phenotype in colon cancer has been suggested by several observations but never conclusively demonstrated. An experimental model to clarify this tissue was established by the creation and characterization of a bank of cell lines that differ mainly, if not exclusively, in the degree of alpha 2,6-sialylation. Human colon cancer cell lines SW48 and SW948, normally unable to elaborate the alpha 2,6-sialyl linkage, were transfected with the beta-galactoside alpha 2,6-sialyltransferase (alpha 2,6ST) cDNA driven by the cytomegaloviral promoter and screened for cell surface alpha 2,6-sialylated sugar chains using fluorescein isothiocyanate-conjugated Sambucus nigra agglutinin (SNA-FITC). A panel of SNA-FITC-positive clones was established that expresses alpha 2,6ST activity of varying degrees. Only the SNA-FITC-positive recombinants express the 1.2 Kb mRNA predicted to be generated from the transfected sequence. No 4.3-4.7 Kb transcripts that are indicative of transcription from the native alpha 2,6ST gene were detected.

Publication types

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

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Colonic Neoplasms
  • Cytomegalovirus
  • DNA Primers
  • DNA, Complementary
  • Fluorescein-5-isothiocyanate
  • Fluorescent Dyes
  • Glycoconjugates / analysis
  • Glycoconjugates / biosynthesis
  • Humans
  • Lectins
  • Molecular Sequence Data
  • Plant Lectins*
  • Polymerase Chain Reaction
  • Promoter Regions, Genetic
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis
  • Rats
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / metabolism
  • Ribosome Inactivating Proteins
  • Sialic Acids / analysis*
  • Sialyltransferases / biosynthesis
  • Sialyltransferases / metabolism*
  • Transfection
  • Tumor Cells, Cultured

Substances

  • DNA Primers
  • DNA, Complementary
  • Fluorescent Dyes
  • Glycoconjugates
  • Lectins
  • Plant Lectins
  • RNA, Messenger
  • Recombinant Proteins
  • Sambucus nigra lectins
  • Sialic Acids
  • Sialyltransferases
  • beta-D-galactoside alpha 2-6-sialyltransferase
  • Ribosome Inactivating Proteins
  • Fluorescein-5-isothiocyanate