Differential expression of alpha2,6-sialyltransferase in colon tumors recognized by a monoclonal antibody.
It has been reported that alpha2,6-sialyltransferase (alpha2,6-ST; E.C. 2.4.99.1) activity is associated with cellular differentiation. To define its role in colon carcinoma differentiation, we have generated murine monoclonal antibodies (MAb) against alpha2,6-sialyltransferase. The MAb, designated 6B9 of IgM isotype, showed strong reactivity with the purified and crude alpha2,6-ST by ELISA and dot blot assays. Western blotting with MAb 6B9 identified purified alpha2,6-ST of MW 47 kDa and the same MW protein from rat and human liver extracts. The MAb also reacted with two other liver proteins of approximate MW 65 and 100 kDa. Immunoperoxidase studies with formalin-fixed paraffin-embedded tissues showed that MAb 6B9 reacts with liver tissues, the staining of hepatocytes was granular and cytoplasmic. There was a distinct pattern of zonal distribution of this enzyme in hepatocytes located particularly in the portal areas of the liver corresponding to zone 1. Normal colon (100%) and hyperplastic polyps (100%) showed very weak to no reactivity. Adenomas (100%) demonstrated moderate reactivity, while the poor (33%), moderate (100%) and well-differentiated (80%) colon adenocarcinomas showed strong reactivity. Results suggest that alpha2,6-ST is associated with the differentiation state of colon tumors.
Gangopadhyay
A
A
Laboratory of Cancer Biology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA.
Perera
S P
SP
Thomas
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P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D015153:Blotting, Western; D003106:Colon / Q000201:enzymology / Q000473:pathology; D003110:Colonic Neoplasms / Q000201:enzymology*; D003111:Colonic Polyps / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D008099:Liver / Q000201:enzymology; D051379:Mice; D051381:Rats; D012799:Sialyltransferases / Q000032:analysis* / Q000276:immunology; D016707:Tissue Fixation; D014407:Tumor Cells, Cultured; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D000911:Antibodies, Monoclonal; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase