Cellular uptake and ex vivo urothelial penetration by oligodeoxynucleotides for optimizing treatment of transitional cell carcinoma.
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.
Bolenz
Christian
C
Department of Urology, Mannheim Medical Center, University of Heidelberg, Mannheim, Germany. christian.bolenz@uro.ma.uni-heidelberg.de
Trojan
Lutz
L
Gabriel
Ute
U
Honeck
Patrick
P
Wendt-Nordahl
Gunnar
G
Schaaf
Axel
A
Alken
Peter
P
Michel
Maurice Stephan
MS
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*
D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D016650:Fluorescein-5-isothiocyanate