(Sialyl)Lewis antigen expression on glycosphingolipids, N- and O-glycans in colorectal cancer cell lines is linked to a colon-like differentiation program

Alterations in the glycomic profile are a hallmark of cancer, including colorectal cancer (CRC). While, the glycosylation of glycoproteins and glycolipids has been widely studied for CRC cell lines and tissues, a comprehensive overview of CRC glycomics is still lacking due to the usage of different samples and analytical methods. In this study, we compared glycosylation features of N-, O- and glycosphingolipid (GSL) glycans for a set of 22 CRC cell lines, all measured by porous graphitized carbon nano-liquid chromatography-tandem mass spectrometry (PGC-nanoLC-MS/MS). An overall, high abundance of (sialyl)Lewis antigens for colon-like cell lines was found while undifferentiated cell lines showed high expression of H blood group antigens and α2-3/6 sialylation. Moreover, significant associations of glycosylation features were found between the three classes of glycans, such as (sialyl)Lewis and H blood group antigens. Integration of the datasets with transcriptomics data revealed positive correlations between (sialyl)Lewis antigens, the corresponding glycosyltransferase (GT) FUT3 and transcription factors (TFs) CDX, ETS, HNF1/4A, MECOM and MYB. This indicates a possible role of these TFs in the upregulation of (sialyl)Lewis antigens, particularly on GSL glycans, via FUT3/4 expression in colon-like cell lines. In conclusion, our study provides insights into the possible regulation of glycans in CRC and can serve as a guide for the development of diagnostic and therapeutic biomarkers.