In vivo gene transfer methods into bladder without viral vectors

目的基因治疗已成为治疗癌症的新策略。当考虑将其应用于膀胱癌时，安全、有效和可重复的体内基因转移方法是必不可少的。在这里，我们研究了三种体内基因转移方法，没有病毒vectors.Materials和方法血凝病毒日本（HVJ）-脂质体具有膜融合活性的腔内注射到大鼠膀胱。用DNA包被的金微载体，用粒子枪轰击兔膀胱粘膜。在兔膀胱粘膜下注射DNA溶液后，通过在针电极之间产生的脉冲直流电（0.15- 0.2A，50 msec，重复8次）检查电转染。结果HVJ-脂质体能有效转染尿道上皮浅层，表达高峰出现在第5天。粒子枪在尿道上皮深层产生不均匀但有效的转染。电转染法可同时转染粘膜下间质细胞和尿道上皮细胞。结论HVJ脂质体是一种潜在的原位癌治疗药物。随着进一步的改进，后两种方法可能适用于治疗局部膀胱肿瘤的辅助治疗。

objective GEne therapy has been adapted as a new strategy for treating cancers. When considering this application to bladder cancer, safe, efficient and repeatable in vivo geno transfer metods are essential. Here, we examined three in vivo gene transfer methods without viral vectors.Materials and methods Hemagglutinating virus of Japan (HVJ)-liposomes possessing the membrane fusion activity were intraluminally injected into rat bladders. Using a particle gun, rabbit bladder mucosa was bombarded with DNA-Coated gold microcarriers. Electrotransfection was also examined in rabbit bladder by pulse direct currents (0.15-0.2 A, 50msec, repeated 8 times) generated between needle electrodes after submucous injection of DNA solution. beta-galactosidase gene and chloramphenicol acetyl-transferase (CAT) gene were used as marker genes.Results HVJ-liposomes efficiently transfected superficial layrs of urothelium with a peak of expression on day 5. The particle gun produced non-uniform but efficient transfection in deeper layrs of the urothelim. By electransfection, submucous interstitial cells were transfected as well as urothelim. No major complications were observed in these three metheds.Conclusion HVJ-liposomes are potentially useful for treating carcinoma in situ. With further refinement, the latter two methods may be suitable for adjuvant therapies in treating localized bladder tumors.