title: Glycosylation of serum ribonuclease 1 indicates a major endothelial origin and reveals an increase in core fucosylation in pancreatic cancer.
abstract: Human pancreatic ribonuclease 1 (RNase 1) is a glycoprotein expressed mainly by the pancreas and also found in endothelial cells. The diagnosis of pancreatic cancer (PaC) remains difficult and therefore the search for sensitive and specific markers is required. Previous studies showed that RNase 1 from human healthy pancreas contained only neutral glycans, whereas RNase 1 from PaC cell lines contained sialylated structures. To determine whether these glycan tumor cell-associated changes were also characteristic of serum RNase 1 and could be used as a marker of PaC, we have analyzed the glycosylation of serum RNase 1. The origin of serum RNase 1 was also investigated. Serum RNase 1 from two PaC patients and two controls was purified and the glycans analyzed by high-performance liquid chromatography (HPLC)-based sequencing and mass spectrometry. Although normal and tumor serum RNase 1 contained the same glycan structures, there was an increase of 40% in core fucosylation in the main sialylated biantennary glycans in the PaC serum RNase 1. This change in proportion would be indicative of a subset of tumor-associated glycoforms of RNase 1, which may provide a biomarker for PaC. Two-dimensional electrophoresis of the RNase 1 from several endothelial cell lines, EA.hy926, human umbilical vein endothelial cells (HUVEC), human mammary microvessel endothelial cells (HuMMEC), and human lung microvessel endothelial cells (HuLEC), showed basically the same pattern and was also very similar to that of serum RNase 1. RNase 1 from EA.hy926 was then purified and presented a glycosylation profile very similar to that from serum RNase 1, suggesting that endothelial cells are the main source of this enzyme.
authors.affiliation: Unitat de Bioquímica i Biologia Molecular, Departament de Biologia, Universitat de Girona, Campus de Montilivi, Girona 17071, Spain.
authors.forename: Sílvia
authors.initials: S
authors.lastname: Barrabés
authors.forename: Lluís
authors.initials: L
authors.lastname: Pagès-Pons
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Radcliffe
authors.forename: Glòria
authors.initials: G
authors.lastname: Tabarés
authors.forename: Esther
authors.initials: E
authors.lastname: Fort
authors.forename: Louise
authors.initials: L
authors.lastname: Royle
authors.forename: David J
authors.initials: DJ
authors.lastname: Harvey
authors.forename: Michel
authors.initials: M
authors.lastname: Moenner
authors.forename: Raymond A
authors.initials: RA
authors.lastname: Dwek
authors.forename: Pauline M
authors.initials: PM
authors.lastname: Rudd
authors.forename: Rafael
authors.initials: R
authors.lastname: De Llorens
authors.forename: Rosa
authors.initials: R
authors.lastname: Peracaula
mesh_terms: D002240:Carbohydrate Sequence; D015180:Electrophoresis, Gel, Two-Dimensional; D004730:Endothelium, Vascular / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D005643:Fucose / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D009439:Neuraminidase / Q000378:metabolism; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D010190:Pancreatic Neoplasms / Q000201:enzymology*; D012016:Reference Values; D012259:Ribonuclease, Pancreatic / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D009844:Oligosaccharides; D005643:Fucose; D012259:Ribonuclease, Pancreatic; D006026:Glycoside Hydrolases; D009439:Neuraminidase