title: Gene transfer to the rat biliary tract with the HVJ-cationic liposome method.
abstract: OBJECTIVE
  The ability to transfer foreign genes into the biliary tract would be useful for the treatment of biliary tract diseases, including cancer, cystic fibrosis and other genetic diseases. To introduce a foreign gene precisely into the rat biliary epithelial cells, we developed a new technique, inserting a polyethylene catheter into the common bile duct through the papilla of Vater by use of a fusigenic cationic liposome with hemagglutinating virus of Japan (HVJ-cationic liposome).
  
  
  METHODS
  Transfection efficiency was estimated with the use of FITC-oligonucleotides (FITC-ODNs) and cDNA of beta-galactosidase (pCAG-lacZ).
  
  
  RESULTS
  FITC-ODNs encapsulated in HVJ-cationic liposome were effectively transfected into cell nuclei of human cholangiocellular carcinoma in vitro after a 30-min incubation as compared with the simple application of naked FITC-ODNs. After in vivo injection of FITC-ODNs using the HVJ-cationic liposome method through the papilla of Vater, fluorescence accumulation was observed only in the epithelial cells of the biliary tract, but not in the parenchymal cells of the liver. Beta-galactosidase expression was observed in the biliary epithelial cells 3 days after the transfection of pCAG-lacZ and was also detected at 14 days, but not at 28 days, without obvious cytotoxicity.
  
  
  CONCLUSIONS
  HVJ-cationic liposome-mediated gene transfer to the biliary tract via the papilla of Vater is a minimally-invasive and an effective gene-delivery method for site-specific targeting to the epithelial cells of the biliary tract, which could be applied to the treatment of human biliary tract diseases.
authors.affiliation: Department of Gastroenterological Surgery, Kyoto University, Graduate School of Medicine, Japan. tetsuzo@kuhp.kyoto-u.ac.jp
authors.forename: T
authors.initials: T
authors.lastname: Uehara
authors.forename: K
authors.initials: K
authors.lastname: Honda
authors.forename: E
authors.initials: E
authors.lastname: Hatano
authors.forename: R
authors.initials: R
authors.lastname: Terao
authors.forename: Y
authors.initials: Y
authors.lastname: Iimuro
authors.forename: N
authors.initials: N
authors.lastname: Yamamoto
authors.forename: M
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Kaneda
authors.forename: Y
authors.initials: Y
authors.lastname: Yamaoka
mesh_terms: D000818:Animals; D001650:Bile Duct Neoplasms; D002404:Catheterization; D002467:Cell Nucleus; D018281:Cholangiocarcinoma; D003135:Common Bile Duct; D018076:DNA, Complementary; D016650:Fluorescein-5-isothiocyanate; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D008081:Liposomes; D008297:Male; D009838:Oligodeoxyribonucleotides; D051381:Rats; D017208:Rats, Wistar; D018108:Respirovirus / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D001616:beta-Galactosidase; D016650:Fluorescein-5-isothiocyanate