人肺癌裸小鼠模型活体成像的动态观察讲述.doc

人肺癌裸小鼠模型活体成像的动态观察 闫明霞1，吴海燕2，朱淼鑫1，刘蕾1，荚德水1，孔韩卫1，姚明1 （1.上海市肿瘤研究所实验病理研究室，上海 200032； 2.扬州大学，扬州 225009） [摘要] 目的：建立稳定表达绿色荧光蛋白的人肺癌细胞系，并探讨小动物活体荧光成像系统在皮下移植瘤模型中的应用。方法：应用慢病毒转染的方法建立表达绿色荧光蛋白的人肺癌细胞系NCI-H460-GFP,并接种至裸小鼠体内建立皮下移植瘤模型，通过小动物活体成像系统连续5周观察肿瘤在动物皮下的动态生长情况。结果：建立了转染率接近100%的NCI-H460-GFP细胞系，在体外及裸小鼠体内均能够长期稳定表达绿色荧光蛋白。活体荧光成像观察发现：1-4周随着肿瘤体积逐渐增大，平均荧光量子数也逐渐增加，5周时随着肿瘤出现明显坏死，平均荧光量子数呈现下降趋势。结论：稳定表达绿色荧光蛋白的NCI-H460-GFP细胞系及其动物模型可以为肺癌研究提供理想的实验材料，应用小动物活体成像系统能够客观定量评价肿瘤在动物体内的生长情况。 [关键词] 人肺癌；活体成像系统；裸小鼠；绿色荧光蛋白;慢病毒载体 Dynamic observation of in vivo biofluorescence imaging of GFP-transfected human lung carcinoma in nude mice YAN Ming-xia1, Wu Hai-yan2,Zhu Miao-xin1,Liu Lei1, Jia De-shui1,Kong Han-wei1, YAO Ming1 (1. Department of Experimental Pathology，Shanghai Cancer Institute，Shanghai 200032，China；2. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China) [ABSTRACT] Objective：To establish a human lung carcinoma cell line, which can stably express green fluorescent protein, and discuss the application of in vivo biofluorescent imaging system in animal model. Methods：Human lung carcinoma NCI-H460 cells were tranfected with gfp gene by lentiviral vector, and they were transplanted into the subcutaneous of the nude mice for preparing the tumor-bearing mice. The biofluorescent signals were detected in sequential five weeks using in vivo fluorescent imaging system. Results：We had successfully built up a human lung carcinoma NCI-H460-GFP cell line, which can display stable high-level GFP expression in vitro and in vivo. In vivo fluorescent images showed that the fluorescent intensities gradually increased with the increase of tumor volume in nude mice from one week to four weeks, whereas fluorescent intensities didnt agree with caliper-measured tumor volume in the fifth week. Conclusions：This high GFP-expressing cell line and animal model may be useful for the study of lung carcinoma, and in vivo fluorescent imaging system was fitted for evaluation and quantitation of the t