Altered expression of sialylated carbohydrate antigens in HT29 colonic carcinoma cells.
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.
Mack
D R
DR
Department of Pediatrics, University of Nebraska Medical Center, Omaha 69198, USA. drmack@unmc.edu
Cheng
P W
PW
Perini
F
F
Wei
S
S
Hollingsworth
M A
MA
D000230:Adenocarcinoma / Q000276:immunology* / Q000473:pathology; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D018395:CA-19-9 Antigen / Q000276:immunology* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003110:Colonic Neoplasms / Q000276:immunology* / Q000473:pathology; D019084:Fluorescent Antibody Technique, Indirect; D005690:Galactose; D005947:Glucose; D019073:HT29 Cells; D006801:Humans; D012799:Sialyltransferases / Q000378:metabolism
D000951:Antigens, Neoplasm; D018395:CA-19-9 Antigen; C042317:DU-PAN-2 antigen, human; D012799:Sialyltransferases; D005947:Glucose; D005690:Galactose