[Cancer Research 49, 945-950, February 15, 1989]
© 1989 American Association for Cancer Research
Oncodevelopmental Expression of —GlcNAcß1–6Man
1-6Manß1— Branched Asparagine-linked Oligosaccharides in Murine Tissues and Human Breast Carcinomas1
J. W. Dennis2 and
S. Laferté
Division of Cancer and Cell Biology, Mount Sinai Hospital Research Institute, Toronto, Ontario M5G 1X5 [J. W. D.]; Department of Medical Genetics, University of Toronto, Toronto, Ontario [J. W. D.]; and Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 0W0 [S. L.], Canada
Increased —GlcNAcß1–6Man
1-6Manß— branching in asparagine-linked oligosaccharides has been observed in murine and human tumor cells and has recently been linked to enhanced metastatic potential in experimental tumor models. Leukoagglutinin (L-PHA) requires the ß1–6-linked lactosamine antenna (ß1–6 branch) for high affinity binding and was used in this study to quantitate these structures on glycoproteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Normal rodent tissues and cell lines were used to standardize the experimental conditions required to quantitate ß1–6-branched oligosaccharide structures and the glycosyltransferase activity which initiates the synthesis of the antenna, ß1–6 N-acetylglucosamine (GlcNAc)-transferase V (EC 2.4.1.155). Secondly, the levels of L-PHA-reactive oligosaccharide were compared in a series of benign and malignant human breast biopsies. Normal human breast tissue and benign lesions showed low expression but 50% of the primary malignancies examined showed significantly elevated L-PHA reactivity. GlcNAc transferase V activities in the human breast carcinomas and in normal murine tissues correlated with the levels of L-PHA reactive oligosaccharide in the tissues. GlcNAc transferase V showed similar ranges of activities, differing by approximately 5-fold between high and low expressing mouse tissues; fibroblasts with and without an activated H-ras oncogene; and low and high expressing human breast carcinomas. The results show that ß1–6 branching in asparagine-linked oligosaccharides is dependent on tissue-specific regulation of GlcNAc transferase V activity. Secondly, a subset of human breast malignancies showed elevated levels of ß1–6-branched oligosaccharides compared to benign samples, suggesting that further studies are warranted to determine whether the presence of these oligosaccharides is associated with metastatic disease and reduced patient survival time.
1 This work was supported by the National Institute of Canada to J. W. D., a Research Scholar of the National Cancer Institute of Canada.
2 To whom requests for reprints should be addressed, at Division of Cancer and Cell Biology, Mt. Sinai Hospital Research Institute, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada.
Received 7/15/88.
Revised 10/ 7/88.
Accepted 11/ 8/88.
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Copyright © 1989 by the American Association for Cancer Research.
