Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells

David R. Mack, Pi-Wan Cheng, Fulvio Perini, Shu Wei, Michael A Hollingsworth

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

To determine whether cell growth conditions impacted carbohydrate expression, HT29 cells were gradually transferred from a conventional glucose-containing media to a glucose-free galactose containing media. Indirect immunofluorescence on acetone fixed cells showed increased expression of sialyl Lewis A antigen (CA19-9), sialyl Lewis C (DUPAN2) and Tn/sialyl-Tn on the surface of HT29 cells grown in the glucose-free galactose containing media compared to those grown in the glucose containing media. Sialyltransferases responsible for the synthesis for these sialylated epitopes were increased in the galactose-fed HT29 cells. Media overlying the cells was subjected to isopycnic ultracentrifugation in cesium chloride and the fractions derived from both glucose and galactose media with equivalent buoyant densities of 1.56 g/L, which are predicted to contain mucin glycoforms, were further separated by HPLC using a Mono-Q anion exchange column. The chromatograph of eluent from the sample derived from the cells growing in the galactose containing media showed an increased peak that reacted with the anti-sialyl Lewis A antibody, CA19-9. These results show that alteration of in vitro culture conditions may cause HT29 colonic carcinoma cells to alter the expression of sialylated carbohydrates.

Original languageEnglish (US)
Pages (from-to)1155-1163
Number of pages9
JournalGlycoconjugate Journal
Volume15
Issue number12
DOIs
StatePublished - Dec 1 1998

Fingerprint

Galactose
HT29 Cells
Cells
Carbohydrates
Carcinoma
Antigens
Glucose
Sialyltransferases
Ultracentrifugation
Cell growth
Mucins
Indirect Fluorescent Antibody Technique
Acetone
Cell culture
Anions
Epitopes
High Pressure Liquid Chromatography
Antibodies
Growth

Keywords

  • Glycoproteins
  • Lewis antigens
  • Mucins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells. / Mack, David R.; Cheng, Pi-Wan; Perini, Fulvio; Wei, Shu; Hollingsworth, Michael A.

In: Glycoconjugate Journal, Vol. 15, No. 12, 01.12.1998, p. 1155-1163.

Research output: Contribution to journalArticle

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