title: (1)H-MRS can detect aberrant glycosylation in tumour cells: a study of the HeLa cell line.
abstract: Glycosylation is the most abundant and diverse form of post-translational modification of proteins. Two types of glycans exist in glycoproteins: N-glycans and O-glycans often coexisting in the same protein. O-glycosylation is frequently found on secreted or membrane-bound mucins whose overexpression and structure alterations are associated with many types of cancer. Mucins have several cancer-associated structures, including high levels of Lewis antigens characterized by the presence of terminal fucose. The present study deals with the identification of MR signals from N-acetylgalactosamine and from fucose in HeLa cells by detecting a low-field signal in one-dimensional (1D) spectra assigned to the NH of N-acetylgalactosamine and some cross peaks assigned to fucose in two-dimensional (2D) spectra. The increase of Golgi pH by treatment with ammonium chloride allowed the N-acetylgalactosamine signal assignment to be confirmed. Behaviour of MR peak during cell growth and comparison with studies from literature taken together made it possible to have more insight into the relationship between aberrantly processed mucin and the presence of non-processed N-acetylgalactosamine residues in HeLa cells. Fucose signals, tentatively ascribed to residues bound to galactose and to N-acetylglucosamine, are visible in both intact cell and perchloric acid spectra. Signals assigned to fucose bound to galactose are more evident in ammonium chloride-treated cells where structural changes of mucin-related Lewis antigens are expected as a result of the higher Golgi pH. A common origin for the N-acetylgalactosamine and fucose resonances attributing them to aberrantly processed mucin can be inferred from the present results.
authors.affiliation: Dipartimento di Tecnologie e Salute and INFN Gruppo Collegato Sanità, Istituto Superiore di Sanità, Viale Regina Elena, Rome, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Palma
authors.forename: Sveva
authors.initials: S
authors.lastname: Grande
authors.forename: Antonella
authors.initials: A
authors.lastname: Rosi
authors.forename: Anna Maria
authors.initials: AM
authors.lastname: Luciani
authors.forename: Laura
authors.initials: L
authors.lastname: Guidoni
authors.forename: Vincenza
authors.initials: V
authors.lastname: Viti
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000378:metabolism; D002452:Cell Count; D005643:Fucose / Q000378:metabolism; D005688:Galactosamine / Q000378:metabolism; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism*; D011522:Protons; D013997:Time Factors
chemical_list: D000951:Antigens, Neoplasm; D011522:Protons; D005643:Fucose; D005688:Galactosamine; D000117:Acetylglucosamine