人结肠腺癌细胞HCT－8绿色荧光标记肿瘤模型的建立

目的筛选表达绿色荧光蛋白基因的人的单克隆结肠腺癌细胞系，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chicken β－acfin-GFP—NEO转染人结肠腺癌细胞HCT－8，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。BALB／CA－nu裸鼠皮下接种1×10^6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的人结肠腺癌细胞株，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。结论绿色荧光蛋白能够在人结肠腺癌细胞HCT-8中长期稳定表达，用绿色荧光蛋白标记的人结肠腺癌细胞HCT-8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。

The Establishment of Human Colon Cancer Model Labeled by GFP

Objective To establish a GFP labeling human Colon cancer cell line and to generate a noninvasive model for monitoring tumor growth in vivo.Metbod Human Colon cancer cells were transfected with chicken β-actin-GFP-NEO by lipofectin 2000 reagent and screened with G418 to produce the stable GFP-expressing clone. Then the labeled HCT-8 cells were implanted into the subcutaneous tissue of nude mice and the growth of the implanted tumor was observed and growth curve of the labeled HCT-8 ceils was analyzed with the whole-body optical imaging system. Result The stable GFP-expressing HCT-8 cell lines were obtained, the growth of the tumor was observed after subcutaneous inoculation of labeled HCT-8 to BALB/AC-nu. Conclusion A GFP-labeled HCT-8 cell line and a mouse tumor model based on the cell line were established, which is a convenient and useful in vivo model for the research on human colon cancer and drug discovery.