Sialyl-dimeric Lewis-X antigen expressed on mucin-like glycoproteins in colorectal cancer metastases.
Colorectal primary carcinomas and metastases from 20 Dukes' stage C or D patients were examined for the immunohistochemical localization and contents of various fucosylated N-acetyl-lactosamine oligomers by specific monoclonal antibodies (MAbs). MAbs used were SH1, specific for Lewis X antigen; FH4, specific for dimeric Lewis X antigen; FH6, specific for sialyl-dimeric Lewis X antigen; and KH1, specific for Lewis Y-Lewis X antigen. The distribution of the carbohydrate antigens identified by these MAbs was heterogeneous within the primary tumor as well as within the metastatic lesion. Examinations of serial sections indicated that areas within an individual tumor which were stained with one MAb were not always reactive with the other MAbs, although these four MAbs identify closely related structures. The degree of MAb reactivity with carcinoma sections was classified by percentage positive carcinoma cells, and primary tumors and metastases from the same patients were compared. An equivalent or higher proportion of carcinoma cells in the metastatic lesions were reactive with MAb FH6 than in the primary colon carcinomas, but each correlation was not seen with the other MAbs. Electrophoretic separation of tumor tissue extracts followed by staining with these MAbs revealed that a component having an approximate molecular weight of 1,000,000 is the major site for the binding of MAbs, FH6, FH4, and KH1. The electrophoretic mobility of the antigenic molecule on polyacrylamide gels as shown by direct MAb bindings was slightly different from that of a major sialomucin revealed by wheat germ agglutinin in the same tissues. MAb FH6 binding to a high molecular weight component was eliminated by prior treatment of the glycoprotein with mild acid or sialidase to remove sialic acid. Simultaneously, binding of MAb SH2, specific for dimeric Lex antigen, to this component increased. An extract was prepared from a liver metastasis, and high molecular weight components were isolated by gel filtration and then fractionated by DEAE-cellulose ion exchange chromatography. A fraction eluted from DEAE-cellulose between 0.10-0.25 M sodium chloride contained most of the MAb FH6 reactivity, as shown by antibody affinity chromatography. These results support a hypothesis that high molecular weight glycoproteins produced by colorectal carcinoma tissues are heterogeneous with regard to their carbohydrate chains and their antigenic structures may change during tumor progression.
Matsushita
Y
Y
Department of Tumor Biology, University of Texas M.D. Anderson Cancer Center, Houston.
Cleary
K R
KR
Ota
D M
DM
Hoff
S D
SD
Irimura
T
T
D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D002240:Carbohydrate Sequence; D002852:Chromatography, Ion Exchange; D003110:Colonic Neoplasms / Q000473:pathology*; D005260:Female; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D016256:Lewis X Antigen / Q000032:analysis* / Q000302:isolation & purification; D046911:Macromolecular Substances; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009077:Mucins / Q000032:analysis*; D009362:Neoplasm Metastasis / Q000473:pathology*; D012004:Rectal Neoplasms / Q000473:pathology*; D012044:Regression Analysis; D012794:Sialic Acids / Q000032:analysis
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D016256:Lewis X Antigen; D046911:Macromolecular Substances; D009077:Mucins; D012794:Sialic Acids