title: Cellular uptake and ex vivo urothelial penetration by oligodeoxynucleotides for optimizing treatment of transitional cell carcinoma.
abstract: OBJECTIVE
  To evaluate cellular uptake and urothelial penetration of oligodeoxynucleotides (ODNs) in transitional cell carcinoma (TCC) cell lines and in a porcine ex vivo model, respectively.
  
  
  METHODS
  A panel of human TCC cell lines (RT 112, HT 1197 and UM-UC3) were exposed tofluorescein-labeled ODNs. Transfection rates were assessed byfluorescence microscopy and fluorescence-activated cell sorting (FACS). Intravesical treatment with ODNs was performed in a porcine ex vivo model. Urothelial penetration was evaluated using fluorescence microscopy of cryosections.
  
  
  RESULTS
  Treatment with ODNs provided transfection rates of at least 96.8% of TCC cells, irrespective of use of a transfection agent. Effective urothelial penetration by ODNs was detected when compared with controls (p = 0.0325). The addition of a liposomal transfection agent significantly increased the penetration depth, allowing affection of deep urothelial cell layers (p = 0.0082).
  
  
  CONCLUSIONS
  High transfection rates of ODNs can be achieved in TCC cells. Urothelial penetration of ODNs was observed down to the deepest cell layers when a transfection agent is added, suggesting a high potential for complementing the chemoresection effects on residual tumor areas during intravesical therapy of non-muscle-invasive TCC.
authors.affiliation: Department of Urology, Mannheim Medical Center, University of Heidelberg, Mannheim, Germany. christian.bolenz@uro.ma.uni-heidelberg.de
authors.forename: Christian
authors.initials: C
authors.lastname: Bolenz
authors.forename: Lutz
authors.initials: L
authors.lastname: Trojan
authors.forename: Ute
authors.initials: U
authors.lastname: Gabriel
authors.forename: Patrick
authors.initials: P
authors.lastname: Honeck
authors.forename: Gunnar
authors.initials: G
authors.lastname: Wendt-Nordahl
authors.forename: Axel
authors.initials: A
authors.lastname: Schaaf
authors.forename: Peter
authors.initials: P
authors.lastname: Alken
authors.forename: Maurice Stephan
authors.initials: MS
authors.lastname: Michel
mesh_terms: D000818:Animals; D002295:Carcinoma, Transitional Cell / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000493:pharmacokinetics*; D013552:Swine; D014162:Transfection / Q000379:methods; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D019459:Urothelium / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D016650:Fluorescein-5-isothiocyanate