UpIb启动子-Smac重组体促膀胱癌细胞凋亡的实验动物研究  

pcDNA3.1 Containing the Smac Gene Controlled by the UpIb Promoter Increases Apoptosis of Bladder Cancer Cells in Animal Models

作　　者：廖贵益[1] 曾甫清[2] 王竞[2] 顾朝辉[2] 王智宇[2] 

机构地区：[1]安徽医科大学第一附属医院泌尿外科,合肥市230022 [2]华中科技大学同济医学院附属协和医院泌尿外科

出　　处：《中国肿瘤临床》2008年第7期398-400,共3页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金资助(编号:30271301)~~

摘　　要：目的:探讨携带Smac基因的膀胱癌细胞特异性表达质粒pcDNA3.1-UpIb promoter-Smac用脂质体包裹后注射到实验动物瘤体内的表达及促癌细胞凋亡的效应。方法:将人膀胱癌细胞株BIU-87注射到Balb/c-nu/nu雄性裸鼠的皮下,构建膀胱癌皮下移植瘤模型。当移植瘤长至直径约0.5cm时,用脂质体将pcDNA3.1-UpIb promoter-Smac包裹后隔天1次连续3次注射到瘤体内,间隔1天后,连续3天瘤体内注射适量丝裂霉素C诱导凋亡。设三组对照:单用脂质体质粒组、单用丝裂霉素C组和空白组。RT-PCR检测瘤组织Smac mRNA的表达,免疫组织化学法检测瘤组织Smac蛋白的表达,苏木素伊红染色镜检、DNA Ladder带凝胶电泳分析和TUNEL-荧光标记检测观察瘤组织癌细胞的凋亡情况。结果:膀胱癌裸鼠移植瘤模型构建成功。瘤体内注射脂质体包裹的pcDNA3.1-UpIbpro-moter-Smac后,与未注射脂质体质粒的对照组比较,Smac的表达增强约2.1倍;先注射脂质体质粒后丝裂霉素C处理和单用丝裂霉素C处理的瘤组织均可见典型的凋亡细胞,但前者的凋亡率为49.8%,显著高于后者的23.5%(P<0.01)。结论:膀胱癌裸鼠移植瘤模型瘤体内直接注射脂质体包裹的pcDNA3.1-UpIb promoter-Smac后,所携带的Smac基因能够在瘤组织中有效表达,并对丝裂霉素C诱导的瘤细胞凋亡有显著的促进作用。该质粒配合丝裂霉素C等凋亡诱导药物的应用,可望为膀胱癌的靶向基因治疗提供一新的治疗模式。Objective： To investigate the expression of the Smac gene in bladder cancer ceils after transfection of a peDNA3.1 vector containing the Smac gene controlled by the UpIb promoter and to observe its effect on bladder cancer cell growth. Methods： Human bladder cancer-derived BIU-87 cells were inoculated into male Balb/c-nu/nu mice subcu- taneously to establish a nude mouse model with bladder cancer. When the injected cells formed a mass that reached a diameter of 0.5 cm, a mixture of liposome and pcDNA3-UpIb promoter-Smac was injected into the tumor in vivo once every other day for 5 days. On days 2 through 4 we induced apoptosis of cancer ceils with daily injections of Mitomycin C into the tumor. There were 3 groups in the study： group 1 was treated with the mixture of liposome and pcDNA3.1-Uplb promoter-Smac, group 2 was treated with Mitomycin C, and group 3 was not given any treatment. The expression of Smac mRNA was measured by RT-PCR. The expression of Smac protein was detected by immunohistochemistry. Apoptosis of cancer cells was observed by H＆E staining, DNA ladder analysis and TUNEL-Fluorescence. Results： The nude mouse model for bladder cancer was successfully established. After intratumoral injection of the liposome and pcDNA3.1-UpIb promoter-Smac mixture, the expression of Smac in the tumors increased 2.1-fold. Apoptosis of the bladder cancer cells was observed in the group treated with the vector combined with Mitomycin C and in the group treated with only Mitomycin C. The apoptosis ratio was 49.8% in the combination group and 23.5% in the mitomycin C alone group. Conelu. sion： Direct injection of the pcDNA3.1-Uplb promoter-Stoat construct into human bladder cancer in nude mice can cause a detectable increase in the expression of the Smac genc in the tumors. This increase subsequently promotes apoptosis which was increased further with the injection of Mitomycin C. Our research lays the foundation for the clinical application of pcDNA3.1-UpIb promoter-Smac combined with apoptosis-inducing r

关 键 词：线粒体促凋亡蛋白 膀眈肿瘤 基因治疗 

分 类 号：R737.14[医药卫生—肿瘤] R73-36[医药卫生—临床医学]