摘要
目的采用阳离子脂质体介导的基因转染的方法对人膀胱移行细胞癌T24细胞进行荧光标记,建立其可在活体荧光成像系统下直接观察肿瘤生长及转移的简便、可靠的原位移植模型。方法以绿色荧光蛋白作为标记基因经阳离子脂质体介导转入人类膀胱移行细胞癌T24细胞株中,经梯度浓度G418筛选获得稳定表达绿色荧光蛋白的细胞克隆并扩大培养,然后接种于裸小鼠膀胱壁内,活体荧光成像系统直接观察原位肿瘤的生长与转移,并与常规石蜡切片苏木素-伊红(HE)染色比较。结果经绿色荧光蛋白标记的人膀胱肿瘤T24细胞在荧光显微镜下发出绿色荧光,并可在体内、外持续稳定表达。绿色荧光蛋白转染后的人膀胱移行细胞癌T24细胞的体内外生物学行为无明显改变,裸小鼠膀胱原位接种5×105个转染后的膀胱肿瘤细胞后1周即可在荧光成像系统下观察到下腹部新生膀胱肿瘤发出绿色荧光,2周后膀胱肿瘤可被触及,4周后经解剖可在荧光成像系统下观察到腹膜后及盆腔淋巴结等远处转移。结论绿色荧光蛋白是新一代的生物源性荧光标记物质,它具有分子量小、对细胞无毒、使用方便及容易检测等优点。绿色荧光蛋白能够在T24人膀胱肿瘤细胞中长期稳定表达,用绿色荧光蛋白标记的人膀胱肿瘤T24细胞建立的可视裸鼠原位移植模型为进一步研究膀胱肿瘤提供了一种简便、可行的新方法。
Objective To label a human bladder cancer cell line and establish a novel human bladder cancer mouse model in which the fluorescent signal of the orthotopic tumor can be detected using the whole-body optical imaging system. Method T24, a human bladder transitional cell carcinoma cell line, were transfected with GFP plasmid to screen stable GFP expressing clones. The latter were implanted into the wall of bladder of the nude mice. The growth, invasion, and metastasis of the tumour were observed and evaluated by the whole-body optical imaging system. The findings were compared with those of HE staining on routine paraffin sections. Result GFP-labeled tumor cells displayed green fluorescence under fluorescent microscopy and showed stable GFP expression in vitro and in vivo. One week after in situ transplantation of 5×10^5 T24 cells, the new bladder cancer could be observed and evaluated by the whole body optical imaging system. Two weeks later, the new bladder tumor could be palpated with hands. Four weeks later, the metastasis to regional drained lymph node in pelvic and retropefitoneal lymph node occurred. The new bladder tumor growth and metastasis were easily observed and accurately evaluated by fluorescent microscope. Conclusion GFP-labeled tumor ceils displayed green fluorescence under fluorescent microscopy and showed stable GFP expression. GFP labeled T24 and the novel human bladder cancer model described here provided a simple and reliable tool for studying the human bladder cancer in vivo.
出处
《热带医学杂志》
CAS
2006年第6期631-634,F0004,共5页
Journal of Tropical Medicine
基金
广东省自然科学基金(No.020059)
关键词
膀胱肿瘤
原位动物模型
绿色荧光蛋白
活体荧光成像系统
bladder carcinoma
orthotopic animal model
green fluorescent protein
whole body optical imaging system
