Elevation of an alpha(1,3)fucosyltransferase activity correlated with apoptosis in the human colon adenocarcinoma cell line, HT-29.
We studied changes in the carbohydrate expression following apoptotic cell death induced by treatment with interferon (IFN)-gamma and anti-Fas antibody using human colon adenocarcinoma HT-29 cells. An apoptotic cell death of HT-29 accompanied with typical DNA fragmentation was observed when the cells, were cultured sequentially with IFN-gamma and anti-Fas antibody. In flow cytometric analyses, the expression of Le(x) and Le(y) antigen was strongly and slightly enhanced, respectively, on the cell surface in accordance with the apoptosis. When the fucosyltransferase (Fuc-T) activities of the lysates from the treated cells were examined relative to those from untreated cells, a 2.5-fold increase of alpha(1,3)-Fuc-T activities and a slight increase of alpha(1,2)-Fuc-T activities were observed, but little or no increase of alpha(1,4)-Fuc-T activity was detected. In Northern blot analyses using probes for Fuc-T III, IV, V, VI and VII genes, strong RNA messages for Fuc-T III, V and/or VI and a weak RNA message for Fuc-T IV were detected in the untreated HT-29 cells. On the other hand, in the treated cells, the messages for Fuc-T III, V and/or VI were found to almost disappear and the 2.3 kb message for Fuc-T IV was observed to elevate 2.8-fold. Therefore, we suggest that the strongly increased expression of Le(x) antigen found on the HT-29 cell surface might be involved in the process of apoptosis, and that the enhancement of the antigen expression seems to result from the increased activity of alpha(1,3)-Fuc-T encoded mainly by the Fuc-T IV gene.
Akamatsu
S
S
Diagnostic Division, Otsuka Pharmaceutical Co. Ltd., Tokushima, Japan.
Yazawa
S
S
Zenita
K
K
Matsumoto
H
H
Tachikawa
T
T
Kannagi
R
R
D000230:Adenocarcinoma / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology*; D000906:Antibodies / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D015152:Blotting, Northern; D002240:Carbohydrate Sequence; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology*; D053938:DNA Fragmentation / Q000187:drug effects; D005434:Flow Cytometry; D005647:Fucosyltransferases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D019014:fas Receptor / Q000276:immunology
D000906:Antibodies; D019014:fas Receptor; D007371:Interferon-gamma; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase