荧光标记技术在肿瘤模型研究中的应用

目的利用绿色荧光小鼠和红色荧光蛋白标记肿瘤细胞,建立荧光标记的小鼠肿瘤模型,并建立活体荧光成像和荧光显微镜成像在整体和细胞水平直接观察肿瘤的技术。方法将小鼠B16黑色素瘤细胞接种到绿色荧光蛋白转基因小鼠皮下,建立GFP小鼠肿瘤模型。以红色荧光蛋白作为标记基因导入小鼠黑色素瘤细胞B16细胞,建立稳定表达红色荧光蛋白的细胞株。将表达红色荧光蛋白B16细胞接种到绿色荧光转基因小鼠皮下,建立双荧光小鼠肿瘤模型。用荧光显微镜和活体荧光成像系统检测小鼠肿瘤的发生发展。结果分别建立了GFP小鼠肿瘤模型和双色荧光小鼠肿瘤模型。利用活体荧光影像仪可以观察双色荧光小鼠模型中受体绿色荧光组织和红色荧光移植肿瘤相互融合。利用荧光显微镜,可以观察到肿瘤内绿色荧光标记的来源于受体小鼠的血管和免疫细胞。经香菇多糖刺激的GFP小鼠肿瘤模型的移植瘤组织中,来源于受体小鼠绿色荧光标记的免疫细胞明显多于经生理盐水刺激的对照小鼠。结论利用绿色荧光小鼠和红色荧光RFP标记肿瘤细胞建立荧光标记的小鼠肿瘤模型,采用活体荧光成像仪和荧光显微镜可在整体和细胞水平直接观察肿瘤的生长以及肿瘤与宿主的相互作用。

Research of Fluorescence-labeled Technique on Tumor Mouse Model

Objective To establish fluorescence-labeled mouse tumor model using GFP transgenic mice and DsRed labeled tumor ceils, and whole-body imaging system and fluorescence microscopy were used to develop the technique of observation for the tumor in the whole-body and cell level directly, Methods B16 melanoma cells were implanted subcutaneously in GFP mouse to copy GFP mouse tumor model, DsRed labeled B16 cells were cloned with G418 selection （B16-RFP）, B16-RFP cells were implanted into the mice subcutaneously to established tumor mouse. The whole-body imaging and fluorescence imaging technique was used to monitor the growth of the implanted tumor. Results A GFP mouse tumor model and a GFP mouse bearing DsRed tumor mouse model were successfully established, Under whole-body imaging scope, the green fluorescence tissue from the host mice and the red in tumor exhibited a fused growth. Under the fluorescence microscopy, the blood vessels and lymphatic cells from host was also exhibited in the tumor tissues. Using this technique and the model, the treatment of Lentinan was directly assayed, which showed that Lentinan significantly increased lymphatic cells and decreased the blood vessels in the tumor tissues compared with saline control. Conclusion We established GFP mouse bearing DsRed tumor models, which could be used for observation the tumor growth and interaction between host cells and tumor cells directly by whole-body imaging system and fluorescence microscopy.