摘要
目的筛选表达绿色荧光蛋白和红色荧光蛋白基因的人的单克隆结直肠癌细胞系,为体内监测肿瘤的早期生长建立一种新的肿瘤动物模型。方法以脂质体2000介导chickenβ-actin-GFP-neo和chickenβ-actin-DsRed-neo转染人结直肠癌细胞HCT-116,经梯度浓度G418筛选获得稳定表达红色和绿色荧光蛋白的细胞克隆并扩大培养。BALB/CA-nu裸鼠皮下接种1×106个发光细胞使其成瘤,活体荧光成像系统观察肿瘤的生长情况。结果获得了稳定表达GFP、DsRed的人结肠癌细胞株,将其接种到裸鼠体内可成瘤,利用活体成像系统观察了肿瘤的生长过程,肿瘤的发光随着观察时间的延长而增加。结论红色和绿色荧光蛋白能够在人结直肠癌细胞HCT-116中长期稳定表达,用红色和绿色荧光蛋白标记的人结直肠癌细胞HCT-116建立的裸鼠肿瘤模型为进一步研究结肠肿瘤和相应的药物筛选提供了一种简便、可行的新方法。
Objective To establish a GFP and DsRed labeling human colorectal cancer cell line and to generatea noninvasive model for monitoring tumor growth in vivo. Methods Human colorectal cancer cells （HCT-116） were transfected with chicken β-actin-GFP-neo and chicken β-actin-DsRed-nen by lipofectin 2000 reagent and screened with G418 to produce the stable GFP and DsRed-expressing clone. Then the labeled HCT-116 cells were implanted into the subcutaneous tissue of nude mice and the growth of the implanted tumor was observed and analyzed with the whole-body optical imaging system. Results The stable GFP and D sRed-expressiug HCT-116 cell lines were obtaioned, subcutaneous to the growth of the tumor was observed after inoculation of the labeled HCT-116 cells below dorsal blank of BALB/CA-nu. Conclusion A GFP and DsRed-labeled HCT-116 cell line and a mouse tumor model based on the cell line were established, which was convenient and useful in vivo model for the research on the human colorectal cancer and drug discovery.
出处
《中国比较医学杂志》
CAS
2009年第9期7-10,I0001,I0002,共6页
Chinese Journal of Comparative Medicine
基金
卫生部项目(200802036)
科技重大专项重大新药创制项目[2009ZX09501-026]
关键词
人结直肠癌细胞
绿色荧光蛋白
红色荧光蛋白
光学影像
小鼠
Human colorectal cancer cell
Green fluorescent protein
Red fluorescent protein
Whole-body optical imaging system
Mouse
