title: Antisense oligodeoxynucleotides directed against aspartyl (asparaginyl) beta-hydroxylase suppress migration of cholangiocarcinoma cells.
abstract: BACKGROUND
  Aspartyl (asparaginyl) beta-hydroxylase (AAH) is an alpha-ketoglutarate-dependent dioxygenase that hydroxylates aspartate and asparagine residues in EGF-like domains of proteins. The consensus sequence for AAH beta-hydroxylation occurs in signaling molecules such as Notch and Notch homologs, which have roles in cell migration.
  
  
  OBJECTIVE
  This study evaluated the potential role of AAH in cell migration using cholangiocarcinoma cell lines as models due to their tendency to widely infiltrate the liver.
  
  
  METHODS
  Five human cholangiocarcinoma cell lines established from human tumors were examined for AAH expression and motility. The effect of antisense oligodeoxynucleotide inhibition of AAH on cholangiocarcinoma cell migration was investigated.
  
  
  RESULTS
  Western blot analysis detected the approximately 86 kDa AAH protein in all five cholangiocarcinoma cell lines, and higher levels of AAH in cell lines derived from moderately or poorly differentiated compared with well-differentiated tumors. Immunocytochemical staining and fluorescence activated cell sorting analysis revealed both surface and intracellular AAH immunoreactivity. Using the phagokinetic non-directional migration assay and a novel ATPLite luminescence-based directional migration assay, we correlated AAH expression with motility. Correspondingly, antisense and not sense or mutated antisense AAH oligodeoxynucleotides significantly inhibited AAH expression and motility in cholangiocarcinoma cells.
  
  
  CONCLUSIONS
  AAH over-expression may contribute to the infiltrative growth pattern of cholangiocarcinoma cells by promoting motility.
authors.affiliation: Departments of Medicine and Pathology, Liver Research Center, Rhode Island Hospital, Brown Medical School, 55 Claverick Street, 4th Floor, Providence, RI 02903, USA.
authors.forename: Takashi
authors.initials: T
authors.lastname: Maeda
authors.forename: Paul
authors.initials: P
authors.lastname: Sepe
authors.forename: Stephanie
authors.initials: S
authors.lastname: Lahousse
authors.forename: Seishu
authors.initials: S
authors.lastname: Tamaki
authors.forename: Munetomo
authors.initials: M
authors.lastname: Enjoji
authors.forename: Jack R
authors.initials: JR
authors.lastname: Wands
authors.forename: Suzanne M
authors.initials: SM
authors.lastname: de la Monte
mesh_terms: D001650:Bile Duct Neoplasms; D001653:Bile Ducts, Intrahepatic; D045744:Cell Line, Tumor / Q000166:cytology / Q000201:enzymology; D002465:Cell Movement / Q000502:physiology*; D018281:Cholangiocarcinoma; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D006899:Mixed Function Oxygenases / Q000235:genetics* / Q000378:metabolism*; D020320:Oligoribonucleotides, Antisense / Q000494:pharmacology
chemical_list: D020320:Oligoribonucleotides, Antisense; D006899:Mixed Function Oxygenases; C058291:aspartic acid 2-oxoglutarate-dependent dioxygenase