兔子膀胱的黏膜下层活体电击 in vivo gene transfer methods in the bladder without viral vectors.pdf

British Journal of Urology (1998), , 870–874 K. HARIMOTO, K. SUGIMURA, C.R. L EE, K. KURATS UKURI and T. KISHIMOTO Department of Urology, Osaka City University Medical School, Osaka, Japan To examine three in vivo gene transfer HVJ liposomes eciently transfected superficial methods, without viral vectors, for use in bladder layers of urothelium, with a peak of expression on day cancer. 5. The particle gun produced a heterogeneous but Three methods were selected: (i) ecient transfection in deeper layers of the urothel- haemagglutinating virus of Japan (HVJ)-liposomes pos- ium. By electrotransfection, both submucosal inter- sessing membrane fusion activity were intraluminally stitial cells and urothelium were transfected. No major injected into rat bladders; (ii) using a particle gun, complications occurred with these three methods. rabbit bladder mucosa was bombarded with DNA- HVJ-liposomes are potentially useful for tre- coated gold microcarriers; (iii) electrotransfection was ating carcinoma in situ. With further refinement, the also assessed in rabbit bladder by pulsed direct currents last two methods may be suitable for adjuvant therapy (0.15–0.2 A, 50 ms, repeated eight times) generated in treating localized bladder tumours. between needle electrodes after the submucosal injec- Gene transfer, bladder, liposome, particle gun, tion of DNA solution. The b-galactosidase gene and electroporation chloramphenicol acetyl-transferase gene were used as marker genes to detect gene transfer.