N‑Linked Glycan Profiling in Neuroblastoma Cell Lines

Although <i>MYCN</i> amplification has been associated with aggressive neuroblastoma, the molecular mechanisms that differentiate low-risk, <i>MYCN</i>-nonamplified neuroblastoma from high-risk, <i>MYCN</i>-amplified disease are largely unknown. Genomic and proteomic studies have been limited in discerning differences in signaling pathways that account for this heterogeneity. N-Linked glycosylation is a common protein modification resulting from the attachment of sugars to protein residues and is important in cell signaling and immune response. Aberrant N-linked glycosylation has been routinely linked to various cancers. In particular, glycomic markers have often proven to be useful in distinguishing cancers from precancerous conditions. Here, we perform a systematic comparison of N-linked glycomic variation between <i>MYCN</i>-nonamplified SY5Y and <i>MYCN</i>-amplified NLF cell lines with the aim of identifying changes in sugar abundance linked to high-risk neuroblastoma. Through a combination of liquid chromatography–mass spectrometry and bioinformatics analysis, we identified 16 glycans that show a statistically significant change in abundance between NLF and SY5Y samples. Closer examination revealed the preference for larger (in terms of total monosaccharide count) and more sialylated glycan structures in the <i>MYCN</i>-amplified samples in comparison to smaller, nonsialylated glycans that are more dominant in the <i>MYCN</i>-nonamplified samples. These results offer clues for deriving marker candidates for accurate neuroblastoma risk diagnosis.