小鼠膀胱癌红色荧光移植瘤模型建立及荧光影像学特点分析  

Red fluorescent transplantation tumor model of mouse bladder carcinoma and fluoroscopic image study

作　　者：杨彬[1] 杨晓峰[1] 付奎[1] 汪海龙[1] 王炜[2] 王晔[2] 

机构地区：[1]山西医科大学附属第一医院泌尿外科,太原030001 [2]浙江大学加州国际纳米技术研究院

出　　处：《中华泌尿外科杂志》2010年第5期331-334,共4页Chinese Journal of Urology

基　　金：山西省科技攻关项目资助（20090321022）

摘　　要：目的 研究红色荧光蛋白（DsRed）标记小鼠膀胱癌生长转移的分子荧光影像学特征.方法 采用脂质体2000介导基因转染法将chickenβ-actin-DsRed-Neo载体转染到小鼠膀胱癌BTT739细胞株;G418筛选后获得稳定表达DsRed的单克隆细胞（BTT739-DsRed）;615小鼠24只随机分3组,后肢皮下注射细胞悬液,第1、2组注射BTT739-DsRed细胞,第3组注射BTT739细胞,建立移植瘤模型.MAESTRO活体成像仪设定激发光波长560～580 nm,发射光波长590～610 nm,曝光时间5000 ms,连续观察4周,第2组每周处死小鼠并剖开成像,记录肿瘤生长转移的荧光影像,测定肿瘤大小及荧光信号值.统计分析肿瘤大小与荧光信号值,全身成像与剖开成像之间的关系.结果 第1周观测到发红色荧光的肿瘤,第2周观测到肿瘤中央局部荧光缺失,病理证实为肿瘤坏死组织及少量结缔组织,第4周观测到局部淋巴结转移,未见远处转移.第1组连续4周肿瘤荧光信号值依次为（89±18）、（122±55）、（133±69）、（715±343）counts;肿瘤大小依次为（13±4）、（45±22）、（83±29）、（253±67）mm2;第2组全身成像4周肿瘤大小依次为（12±3）、（50±23）、（90±29）、（290±74）mm2,剖开成像依次为（12±5）、（72±30）、（141±43）、（524±237）mm2;肿瘤大小与荧光信号强度呈线性正相关（r=0.74,t=3.97,P〈0.05）,全身成像与剖开成像肿瘤大小呈线性正相关（r=0.97,t=10.00,P〈0.05）,全身成像测得的肿瘤大小为剖开后成像的（70.85±17.13）%. 结论小鼠膀胱癌红色荧光移植瘤模型可以直观、连续、敏感地观察肿瘤的生长和转移,随着肿瘤增大,荧光范围也增大,肿瘤发生坏死后荧光消失,肿瘤发生转移后红色荧光表达亦随之转移.Objective To explore molecular fluorescence imaging features of the growth and metastasis of DsRed-marked mouse bladder carcinoma. Methods The study used lipofectamine 2000 transfection method,transferred on the BTT739 cells with plasmid chickenβ-actin-DsRed-Neo vector.The stably expressing BTT739-DsRed monoelonal cells were got with G418 selection.It randomly divided the 615 mouse of 24 into three groups,injected cell suspension on the hindlimb,the first and second group with BTT739-DsRed cell and the third group with BTT739 cell to found xenograft roodel.MAESTRO imager recorded fluorescence images of the growth and metastasis of the tumors in vivo and the fluorescence intensity was measured.The excitation wavelength was 560-580 nm,emission wavelength was 590-610 nm and exposure time was 5000 ms.After continuous observation of 4 weeks,every week killed the mouse of the second group and cut into image,made records of the red fluorescent mouse bladder cancer xenograft model,measured the tumor size and fluorescence sighal values; analyzed the relations between the tumor size and fluorescence signal values as well as between the whole image and cut image. Results DsRed tumor could be observed at the first week. Central local fluorescence loss could be detected at the second week, pathologically confirmed necrotic tumor tissue and a little connective tissue. At the fourth week, a local lymph node metastasis could be observed with no distant metastasis. The measured values of fluorescent signal were as follows： （89±18）, （122±55）, （133±69）, （715±343）counts; the tumor size were as follows： （13±4）, （45±22）, （83±29）, （253±67）mm2. The whole body image of tumor size were as follows： （12± 3）,（50±23）, （90±29）, （290±74）mm2. The cut image of tumor size were as follows： （12±5）, （72±30）, （141±43）, （524±237）mm2. The tumor size and fluorescent signal values reflect positive linear correlation with 0. 74 coefficient （t= 3. 97, P〈0.05）, who

关 键 词：红色荧光蛋白 膀胱肿瘤 全身成像 动物模型 

分 类 号：R737.14[医药卫生—肿瘤]