裸鼠原位非肌层浸润性膀胱癌模型的优选及验证

Optimization and verification of orthotopic non-muscle invasive bladder cancer model in nude mice

目的:比较4种不同的细胞移植方法的优缺点,优选出裸鼠原位非肌层浸润性膀胱癌(NMIBC)模型复制技术,并通过膀胱灌注吉西他滨治疗膀胱癌的疗效对该模型进行验证,为治疗膀胱癌药物的筛选提供稳定高效的动物模型。方法:采用“稀酸碱腐蚀”或“硝酸银烧灼”方式破坏膀胱黏膜后,经尿道膀胱内注入人膀胱移行细胞癌细胞(T24细胞);“手术直视下机械损伤膀胱黏膜”后膀胱内注入T24细胞;“膀胱壁内直接注射T24细胞”进行造模,于造模后14 d处死裸鼠,观察膀胱(肿瘤)质量和组织病理学改变确认原位膀胱癌形成情况。并采用优选到的膀胱壁内直接注射T24细胞的造模方法,观察原位膀胱癌形成的动态变化过程。采用一线膀胱灌注化疗药物吉西他滨验证膀胱壁内直接注射T24细胞复制的模型的适用性。结果:“稀酸碱腐蚀”或“硝酸银烧灼”方式破坏膀胱黏膜后,经尿道膀胱内灌注T24细胞,所有裸鼠膀胱内均未能生长出肿瘤;“手术直视下机械损伤膀胱黏膜”后裸鼠膀胱内注入T24细胞,成瘤率为100%,但肿瘤数量不一,常多点发生。裸鼠膀胱壁内直接注射T24细胞方式,成瘤率为100%,基本仅为1个肿瘤,肿瘤大小一致性良好,并且在15 d内缓慢线性生长,第18天至第31天肿瘤呈快速线性生长。吉西他滨150 mg/kg膀胱灌注可以明显抑制膀胱壁内直接注射T24细胞复制的裸鼠原位NMIBC的生长,抑瘤率达97.1%。结论:“稀酸碱腐蚀”或“硝酸银烧灼”方式破坏膀胱黏膜,经尿道膀胱内灌注T24细胞不能复制原位膀胱癌模型;“手术直视下机械损伤膀胱黏膜”后膀胱内注射T24细胞,形成原位膀胱癌,数量、大小不一;膀胱壁内直接注射T24细胞可成功制备原位NMIBC模型,可以用于原位NMIBC治疗药物的评价。

AIM:To optimize an orthopedic non-muscle invasive bladder cancer(NMIBC)model in nude mouse by comparing four different ways of cellular transplantation,and to evaluate the efficacy of drug by bladder instillation,so as to provide a stable and efficient animal model for the treatment of bladder cancer.METHODS:After disruption of bladder mucosa by dilute acid-alkali or silver nitrate,T24 cells were instilled into the nude mouse bladder.T24 cells were injected directly into the bladder with mechanical injury of bladder mucosa.T24 cells were injected into the bladder wall.On the 14th day after making models,the nude mice were sacrificed.And the bladder mass and histopathological changes of tumor(including bladder)was observe to confirm the formation of orthopedic bladder cancer.The dynamic changes of orthopedic bladder cancer were observed after injecting T24 cells into the bladder wall.Gemcitabine was used to verify the applicability of the model of injecting T24 cells into the bladder wall in vivo.RESULTS:No tumor was found in the bladder after intravesical instillation of T24 cells with dilute acid-alkali or silver nitrate treatment.With mechanical injury of bladder mucosa,all nude mice had tumors after injection T24 cells.But the number of tumors varied and often occurred at multiple sites.The tumor was found in the bladder of all nude mice by injecting T24 cells into bladder wall,and there was only one tumor.The tumor showed slow linear growth within 15 days and rapid linear growth from day 18 to 31.In vivo efficacy evaluation,gemcitabine 150 mg/kg intravesical perfusion could significantly inhibit the growth of NMIBC in nude mice replicated by direct injection of T24 cells into the bladder wall,and the tumor inhibition rate was 97.1%.CONCLUSION:The orthotopic NMIBC model can not be established with the bladder mucosa injuried by dilute acid-alkali or silver nitrate treatment.The number and size of orthotopic bladder cancer are different by mechanical injury of bladder mucosa.Injection of T24 cells into the bladder wall of nude mouse can successfully establish the orthotopic NMIBC model,which can be used for the evaluation of NMIBC therapeutic drugs.