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Glycoconjugate Journal

, Volume 15, Issue 12, pp 1155–1163 | Cite as

Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells

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

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.

glycoproteins mucins Lewis antigens 

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Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • David R. Mack
    • 1
  • Pi-Wan Cheng
    • 2
  • Fulvio Perini
    • 3
  • Shu Wei
    • 1
  • Michael A. Hollingsworth
    • 4
  1. 1.Department of PediatricsUniversity of Nebraska Medical Center and Center for Human NutritionOmahaUSA
  2. 2.Biochemistry and Molecular Biology; Eppley Institute for Research in Cancer and Allied SciencesUniversity of Nebraska Medical Center and Center for Human NutritionOmahaUSA
  3. 3.Eppley Institute for Research in Cancer and Allied SciencesUniversity of Nebraska Medical Center and Center for Human NutritionOmahaUSA
  4. 4.Institute for Research in Cancer and Allied SciencesUniversity of Nebraska Medical Center and Center for Human NutritionOmahaUSA

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