title: The formation of the oncofetal J28 glycotope involves core-2 beta6-N-acetylglucosaminyltransferase and alpha3/4-fucosyltransferase activities.
abstract: The feto-acinar pancreatic protein or FAPP, the oncofetal glycoisoform of bile salt-dependent lipase (BSDL), is characterized by the presence of the J28 glycotope recognized by mAbJ28. This fucosylated epitope is carried out by the O-linked glycans of the C-terminal mucin-like region of BSDL. This glycotope is expressed by human tumoral pancreatic tissues and by human pancreatic tumoral cell lines such as SOJ-6 and BxPC-3 cells. However, it is not expressed by the normal human pancreatic tissues and by MiaPaCa-2 and Panc-1 cells. Due to the presence of many putative sites for O-glycosylation on FAPP and BSDL, the structure of the J28 glycotope cannot be attained by classical physical methods. In the first part of the present study, we have determined which glycosyltransferases were differently expressed in pancreatic tumoral cell lines compared to normal tissues, focusing in part on fucosyltransferases (Fuc-T) and core-2 beta6-N-acetylglucosaminyltransferase (Core2GlcNAc-T). Our data suggested that alpha2-Fuc-T activity was decreased in the four cell lines tested (SOJ-6, BxPC-3, MiaPaCa-2, and Panc-1). The alpha(1-3) and alpha(1-4) fucosylations were decreased in tumor cells that do not express the J28 glycotope whereas alpha4-Fuc-T and Core2GlcNAc-T activities were significantly increased in SOJ-6 cells which best expressed the J28 glycotope. Therefore, we wished to gain information about glycosyltransferases involved in the building of this structure by transfecting the cDNA encoding the mucin-like region of BSDL in CHO-K1 also expressing Core2GlcNAc-T and/or FUT3 and/or FUT7 activities. These CHO-K1 cells have been previously transfected with the cDNA encoding Core2GlcNAc-T and/or FUT3 and/or FUT7. Data indicated that the C-terminal peptide of BSDL (Cter) produced by those cells did not carry out the J28 glycotope unless Core2GlcNAc-T activity is present. Further transfection with FUT3 cDNA, increased the antibody recognition. Nevertheless, transfection with FUT3 or FUT7 alone did not generate the formation of the J28 glycotope on the C-terminal peptide. Furthermore, the Cter peptide produced by CHO-K1 cells expressing Core2GlcNAc-T was more reactive to the mAbJ28 after in vitro fucosylation with the recombinant soluble form of FUT3. These data suggested that the J28 glycotope encompasses structures initiated by Core2GlcNAc-T and further fucosylated by alpha3/4-Fuc-T such as FUT3, likely on GlcNAc residues.
authors.affiliation: INSERM U 260, Unité de Recherche de Physiopathologie des Régulations Hormono-Nutritionnelles, Faculté de Médecine-Timone, 27 Boulevard Jean Moulin, 13385 Marseilles-Cedex 5, France.
authors.forename: L
authors.initials: L
authors.lastname: Panicot
authors.forename: E
authors.initials: E
authors.lastname: Mas
authors.forename: E
authors.initials: E
authors.lastname: Pasqualini
authors.forename: M
authors.initials: M
authors.lastname: Zerfaoui
authors.forename: D
authors.initials: D
authors.lastname: Lombardo
authors.forename: M O
authors.initials: MO
authors.lastname: Sadoulet
authors.forename: A
authors.initials: A
authors.lastname: El Battari
mesh_terms: D000951:Antigens, Neoplasm / Q000096:biosynthesis*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D000939:Epitopes / Q000096:biosynthesis*; D005647:Fucosyltransferases / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis* / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000737:chemistry / Q000201:enzymology; D010446:Peptide Fragments / Q000378:metabolism; D002787:Sterol Esterase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured
chemical_list: D000951:Antigens, Neoplasm; D018076:DNA, Complementary; D000939:Epitopes; D006023:Glycoproteins; D010446:Peptide Fragments; C031192:oncofetal antigens; D005647:Fucosyltransferases; D017351:N-Acetylglucosaminyltransferases; C053024:beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-acetylglucosaminyl transferase; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase; C054784:bile salt-stimulated lipase; D002787:Sterol Esterase