In vivo gene transfer methods in the bladder without viral vectors.
OBJECTIVE
To examine three in vivo gene transfer methods, without viral vectors, for use in bladder cancer.


METHODS
Three methods were selected: (i) haemagglutinating virus of Japan (HVJ)-liposomes possessing membrane fusion activity were intraluminally injected into rat bladders; (ii) using a particle gun, rabbit bladder mucosa was bombarded with DNA-coated gold microcarriers; (iii) electrotransfection was also assessed in rabbit bladder by pulsed direct currents (0.15-0.2 A, 50 ms, repeated eight times) generated between needle electrodes after the submucosal injection of DNA solution. The beta-galactosidase gene and chloramphenicol acetyl-transferase gene were used as marker genes to detect gene transfer.


RESULTS
HVJ liposomes efficiently transfected superficial layers of urothelium, with a peak of expression on day 5. The particle gun produced a heterogeneous but efficient transfection in deeper layers of the urothelium. By electrotransfection, both submucosal interstitial cells and urothelium were transfected. No major complications occurred with these three methods.


CONCLUSIONS
HVJ-liposomes are potentially useful for treating carcinoma in situ. With further refinement, the last two methods may be suitable for adjuvant therapy in treating localized bladder tumours.
Department of Urology, Osaka City University Medical School, Japan.
K
K
Harimoto
K
K
Sugimura
C R
CR
Lee
K
K
Kuratsukuri
T
T
Kishimoto
D000818:Animals; D015415:Biomarkers; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D018274:Electroporation; D018014:Gene Transfer Techniques; D008081:Liposomes / Q000008:administration & dosage*; D008297:Male; D051379:Mice; D011817:Rabbits; D051381:Rats; D018108:Respirovirus; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000628:therapy*; D001616:beta-Galactosidase / Q000378:metabolism
D015415:Biomarkers; D008081:Liposomes; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase