Abstract
Various cancers such as colorectal cancer (CRC) are associated with alterations in protein glycosylation. CRC cell lines are frequently used to study these (glyco)biological changes and their mechanisms. However, differences between CRC cell lines with regard to their glycosylation have hitherto been largely neglected. Here, we comprehensively characterized the N-glycan profiles of 25 different CRC cell lines, derived from primary tumors and metastatic sites, in order to investigate their potential as glycobiological tumor model systems and to reveal glycans associated with cell line phenotypes. We applied an optimized, high-throughput membrane-based enzymatic glycan release for small sample amounts. Released glycans were derivatized to stabilize and differentiate between α2,3- and α2,6-linked N-acetylneuraminic acids, followed by N-glycosylation analysis by MALDI-TOF(/TOF)-MS. Our results showed pronounced differences between the N-glycosylation patterns of CRC cell lines. CRC cell line profiles differed from tissue-derived N-glycan profiles with regard to their high-mannose N-glycan content but showed a large overlap for complex type N-glycans, supporting their use as a glycobiological cancer model system. Importantly, we could show that the high-mannose N-glycans did not only occur as intracellular precursors but were also present at the cell surface. The obtained CRC cell line N-glycan features were not clearly correlated with mRNA expression levels of glycosyltransferases, demonstrating the usefulness of performing the structural analysis of glycans. Finally, correlation of CRC cell line glycosylation features with cancer cell markers and phenotypes revealed an association between highly fucosylated glycans and CDX1 and/or villin mRNA expression that both correlate with cell differentiation. Together, our findings provide new insights into CRC-associated glycan changes and setting the basis for more in-depth experiments on glycan function and regulation.
Highlights
From the ‡Center for Proteomics and Metabolomics, §Department of Surgery, and ¶Department of RheumatologyLeiden University Medical Center, Leiden, The Netherlands; ʈDepartment of Molecular Cell Biology and Immunology and **Division of BioAnalytical Chemistry, VU University Medical Center, Amsterdam, The Netherlands; ‡‡Univ
We investigated the N-glycome of 25 Colorectal cancer (CRC) cell lines including 21 cell lines derived from primary tumors and four from metastases revealing vast glycosylation differences between the cell lines
It was shown that genetic profiles as well as functional markers and morphological features are commonly retained for CRC cell lines [12, 41]
Summary
From the ‡Center for Proteomics and Metabolomics, §Department of Surgery, and ¶Department of RheumatologyLeiden University Medical Center, Leiden, The Netherlands; ʈDepartment of Molecular Cell Biology and Immunology and **Division of BioAnalytical Chemistry, VU University Medical Center, Amsterdam, The Netherlands; ‡‡Univ. Colorectal cancer (CRC) is a very prevalent and heterogeneous pathology with highly variable disease progression and clinical outcome among patients. As CRC is often asymptomatic in the first years, only 40% of the patients are diagnosed at stage I-II, pointing to the urgent need of sensitive diagnostic tools for early detection and effective, curative treatment [3]. In this context, understanding the complex mechanisms of CRC is an over-. N-glycosylation Profiling of Colorectal Cancer Cell Lines riding condition for the development of new, more efficient means of detection, treatment, and prognosis of the disease
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
