Glycoconjugates constitute a major class of biomolecules which include glycoproteins, glycosphingolipids and proteoglycans. Glycans are involved in several physiological and pathological conditions, such as host–pathogen interactions, cell differentiation, migration, tumour invasion and metastisation, cell trafficking and signalling. Cancer is associated with glycosylation alterations in glycoproteins and glycolipids. This review describes various aspects of protein glycosylation with the focus on alterations associated with human cancer. The application of these glycosylation modifications as biomarkers for cancer detection in tumour tissues and serological assays is summarised.

BLAST analysis of expressed sequence tags (ESTs) using the coding sequence of the human UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferase, designated beta4Gal-T1, revealed a large number of ESTs with identical as well as similar sequences. ESTs with sequences similar to that of beta4Gal-T1 could be grouped into at least two non-identical sequence sets. Analysis of the predicted amino acid sequence of the novel ESTs with beta4Gal-T1 revealed conservation of short sequence motifs as well as cysteine residues previously shown to be important for the function of beta4Gal-T1. The likelihood that the identified ESTs represented novel galactosyltransferase genes was tested by cloning and sequencing of the full coding region of two distinct genes, followed by expression. Expression of soluble secreted constructs in the baculovirus system showed that these genes represented genuine UDP-galactose:beta-N-acetylglucosamine beta1, 4-galactosyltransferases, thus designated beta4Gal-T2 and beta4Gal-T3. Genomic cloning of the genes revealed that they have identical genomic organizations compared with beta4Gal-T1. The two novel genes were located on 1p32-33 and 1q23. The results demonstrate the existence of a family of homologous galactosyltransferases with related functions. The existence of multiple beta4-galactosyltransferases with the same or overlapping functions may be relevant for interpretation of biological functions previously assigned to beta4Gal-T1.

The available information favours a greater impact of environmental exposures on intestinal type gastric cancer, and risk factors for the cardia and distal stomach cancers also appear to be different. We aimed to estimate the association between fruit and vegetable intake and gastric cancer, by location and histological type. We performed a population-based case–control study and a meta-analysis of studies addressing this issue. Incident cases (n=305) were identified in two large teaching hospitals (Porto, Portugal), and controls were randomly sampled among city dwellers (n=1129). Published studies were searched through PubMed, and effects were combined with random effects meta-analysis. In our case–control study, the odds ratio (OR) for the comparison of the highest vs. lowest tertile of fruit consumption was 0.47 [95% confidence interval (CI): 0.21–1.05] for cardia, 0.53 (95% CI: 0.35–0.80) for non-cardia cancer, 0.36 (95% CI: 0.20–0.62) for intestinal, and 1.00 (95% CI: 0.53–1.90) for the diffuse histological type. For vegetables, the corresponding OR was 0.59 (95% CI: 0.26–1.35), 0.85 (95% CI: 0.58–1.26), 0.95 (95% CI: 0.57–1.57), and 0.60 (95% CI: 0.32–1.14). In meta-analysis, considering fruit consumption (highest vs. lowest category), the combined OR was 0.58 (95% CI: 0.38–0.89) for cardia, 0.61 (95% CI: 0.44–0.84) for non-cardia, 0.49 (95% CI: 0.33–0.72) for intestinal type, and 0.82 (95% CI: 0.57–1.20) for diffuse type. Vegetables also decreased the risk of cardia (OR=0.63, 95% CI: 0.50–0.79), non-cardia (OR=0.75, 95% CI: 0.59–0.95), intestinal (OR=0.61, 95% CI: 0.44–0.86), and diffuse type (OR=0.67, 95% CI: 0.44–1.01). Fruit or vegetable intake was associated with a decreased risk of gastric cancer regardless of the anatomical location and the histological type, although dietary intake had a more clear-cut protective effect on intestinal type cancers.

In intestinal metaplasia and 30% of gastric carcinomas, MUC2 intestinal mucin and the intestine-specific transcription factors Cdx-1 and Cdx-2 are aberrantly expressed. The involvement of Cdx-1 and Cdx-2 in the intestinal development and their role in transcription of several intestinal genes support the hypothesis that Cdx-1 and/or Cdx-2 play important roles in the aberrant intestinal differentiation program of intestinal metaplasia and gastric carcinoma. To clarify the mechanisms of transcriptional regulation of the MUC2 mucin gene in gastric cells, pGL3 deletion constructs covering 2.6 kb of the human MUC2 promoter were used in transient transfection assays, enabling us to identify a relevant region for MUC2 transcription in all gastric cell lines. To evaluate the role of Cdx-1 and Cdx-2 in MUC2 transcription we performed co-transfection experiments with expression vectors encoding Cdx-1 and Cdx-2. In two of the four gastric carcinoma cell lines and in all colon carcinoma cell lines we observed transactivation of the MUC2 promoter by Cdx-2. Using gel shift assays we identified two Cdx-2 binding sites at -177/-171 and -191/-187. Only simultaneous mutation of the two sites resulted in inhibition of Cdx-2-mediated transactivation of MUC2 promoter, implying that both Cdx-2 sites are active. Finally, stable expression of Cdx-2 in a gastric cell line initially not expressing Cdx-2, led to induction of MUC2 expression. In conclusion, this work demonstrates that Cdx-2 activates the expression of MUC2 mucin gene in gastric cells, inducing an intestinal transdifferentiation phenotype that parallels what is observed both in intestinal metaplasia and some gastric carcinomas.

Mucins are a large family of heavily O-glycosylated proteins that cover all mucosal surfaces and constitute the major macromolecules in most body fluids. Mucins are primarily defined by their variable tandem repeat (TR) domains that are densely decorated with different O-glycan structures in distinct patterns, and these arguably convey much of the informational content of mucins. Here, we develop a cell-based platform for the display and production of human TR O-glycodomains (~200 amino acids) with tunable structures and patterns of O-glycans using membrane-bound and secreted reporters expressed in glycoengineered HEK293 cells. Availability of defined mucin TR O-glycodomains advances experimental studies into the versatile role of mucins at the interface with pathogenic microorganisms and the microbiome, and sparks new strategies for molecular dissection of specific roles of adhesins, glycoside hydrolases, glycopeptidases, viruses and other interactions with mucin TRs as highlighted by examples.