title: Down-regulation of FUT3 and FUT5 by shRNA alters Lewis antigens expression and reduces the adhesion capacities of gastric cancer cells.
abstract: BACKGROUND
  Lewis antigens are fucosylated glycoconjugates involved in the development of several pathologies. The adhesion of sialyl-Lewis antigens to E-selectin is a key step in the development of metastasis and the glycosidic component of CD44 plays a key role in the binding to hyaluronic acid, a component of the extracellular matrix associated to tumor development and invasion. Fucosyltransferases are enzymes that add fucose to precursor glycan structures: FUT3 and FUT5 catalyze the addition of fucose to the α1-3,4 position and are detected in epithelial cells. In this study, we have analyzed the effects of silencing FUT3, FUT5 or FUT3/FUT5, in two gastric cancer cell lines, in the expression of Lewis antigens and in the adhesive and migratory capacities of the cells.
  
  
  METHODS
  FUT3, FUT5 and FUT3/FUT5 were down-regulated using lentiviral delivery of shRNAs in MKN45 and GP220 gastric cancer cells.
  
  
  RESULTS
  In the infected cells, decreased levels of FUT3 and FUT5 mRNA detected by quantitative RT-PCR; and lower levels of sialyl-Lewis antigens, evaluated by flow cytometry, were observed. The adhesion to endothelial cells trough the binding to E-selectin, and the binding to hyaluronic acid were reduced in the shFUT3, shFUT5 and shFUT3/FUT5, whereas the levels of CD44, analyzed by western blot, did not change.
  
  
  CONCLUSIONS
  The down-regulation of FUT3, FUT5 and FUT3/FUT5 reduces the expression of sialyl-Lewis antigens and the adhesion and binding capacities of gastric cancer cells; and allows to identify the specific α1-3,4 fucosyltransferases implicated in the Lewis antigens synthesis in this cellular model.
authors: Programa de Recerca en Càncer, IMIM-Hospital del Mar, Parc de Recerca Biomèdica de Barcelona, Spain.
authors: Mercè
authors: M
authors: Padró
authors: Lara
authors: L
authors: Cobler
authors: Marta
authors: M
authors: Garrido
authors: Carme
authors: C
authors: de Bolós
mesh_terms: D000941:Antigens / Q000378:metabolism*; D015153:Blotting, Western; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D015536:Down-Regulation; D005434:Flow Cytometry; D005647:Fucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D034741:RNA, Small Interfering / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013274:Stomach Neoplasms / Q000276:immunology / Q000378:metabolism / Q000473:pathology*
chemical_list: D000941:Antigens; D034741:RNA, Small Interfering; C553868:FUT5 protein, human; D005647:Fucosyltransferases; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase