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Colorectal cancer cell lines show striking diversity of their O-glycome reflecting the cellular differentiation phenotype

Journal article
Authors K. Madunic
T. Zhang
O. A. Mayboroda
S. Holst
K. Stavenhagen
Chunsheng Jin
Niclas G. Karlsson
G. S. M. Lageveen-Kammeijer
M. Wuhrer
Published in Cellular and Molecular Life Sciences
Pages 14
ISSN 1420-682X
Publication year 2020
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Pages 14
Language en
Keywords O-Glycosylation, Glycomics, Cell lines, Colorectal cancer, Mass, spectrometry, Porous graphitized carbon liquid chromatography, consensus molecular subtypes, histo-blood group, colon-cancer, selectin, ligand, tn antigen, expression, glycosylation, mucin, carcinoma, glycans, Biochemistry & Molecular Biology, Cell Biology
Subject categories Cell biology


Alterations in protein glycosylation in colorectal cancer (CRC) have been extensively studied using cell lines as models. However, little is known about their O-glycome and the differences in glycan biosynthesis in different cell types. To provide a better understanding of the variation in O-glycosylation phenotypes and their association with other molecular features, an in-depth O-glycosylation analysis of 26 different CRC cell lines was performed. The released O-glycans were analysed on porous graphitized carbon nano-liquid chromatography system coupled to a mass spectrometer via electrospray ionization (PGC-nano-LC-ESI-MS/MS) allowing isomeric separation as well as in-depth structural characterization. Associations between the observed glycan phenotypes with previously reported cell line transcriptome signatures were examined by canonical correlation analysis. Striking differences are observed between the O-glycomes of 26 CRC cell lines. Unsupervized principal component analysis reveals a separation between well-differentiated colon-like and undifferentiated cell lines. Colon-like cell lines are characterized by a prevalence of I-branched and sialyl Lewis x/a epitope carrying glycans, while most undifferentiated cell lines show absence of Lewis epitope expression resulting in dominance of truncated alpha 2,6-core sialylated glycans. Moreover, the expression of glycan signatures associates with the expression of glycosyltransferases that are involved in their biosynthesis, providing a deeper insight into the regulation of glycan biosynthesis in different cell types. This untargeted in-depth screening of cell line O-glycomes paves the way for future studies exploring the role of glycosylation in CRC development and drug response leading to discovery of novel targets for the development of anti-cancer antibodies.

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