绿色荧光蛋白转基因小鼠诱导肝癌模型的建立

目的建立绿色荧光蛋白（green fluorescent protein，GFP）转基因小鼠肝癌模型，观察GFP蛋白在诱癌过程中荧光的变化。方法采用二乙基亚硝胺（diethylnitrosamine，DENA）／四氯化碳（CCk，乙醇／DENA诱导肝癌共20周，实验组50只和对照组10只均为GFP转基因小鼠。每周监测小鼠体重的变化；常规HE染色动态观察病理组织学；第4、12、16周处死小鼠取肝组织制作冷冻切片观察荧光表达，并以石蜡HE染色观察病理变化；第20周时处死小鼠取其肝肿物进行原代培养，观察肿瘤细胞中荧光表达和分布。结果实验组GFP转基因小鼠死亡15只，死亡率30％（15／50）。对照组10只小鼠未发生死亡。GFP转基因小鼠在诱癌的第4、12、16、20周依次出现肝炎，肝硬化、癌前病变和癌变等。荧光显微镜下观察到诱癌开始前、第4周、第12周和第16周肝脏组织的冷冻切片有GFP蛋白的表达，诱癌第20周肝肿物原代培养中肿瘤细胞的细胞质和细胞核中有GFP蛋白表达。结论本实验成功建立GFP转基因小鼠肝癌发生的动物模型，可动态观察肝癌细胞中GFP蛋白的表达。

A green fluorescent protein transgenic mouse model of hepatocellular carcinoma

Objective To establish a stable green fluorescent protein （GFP）transgenie mouse model of hepatoeellular carcinmna （HCC）and observe changes in fluorescence during carcinogenesis. Methods GFP transgenic mice were divided into an experimental group（n =50）and a control group（n = 10）.HCC was induced in the experimental group by treatment with diethy]nitrosamine（DENA）/ carbon tetrachloride（CCl4）/ethanol for 20 weeks. Animal body weight was monitored weekly.At weeks 4, 12,and 16,some animals were sacrificed and frozen sections of liver tissue were prepared for fluorescence analysis ,and HE-stained paraffin sections were prepared for observing dynamic histopathologic changes.At week 20,the remaining mice were sacrificed,and the liver tumors were used to establish primary cultures,in which the expression and distribution of fluorescence was studied. Results In the experimental group,30% of mice（ 15/50）died,while no death occurred in the control group.Hepatitis, cirrhosis,precancerous lesions and HCC appeared,respectively,at weeks 4, 12, 16 and 20.GFP protein expression was detected by fluorescence microscopy in frozen sections collected at weeks 4, 12,and 16,as well as in the cytoplasm and nuclei of cultured tumor cells at week 20. Conclusion GFP expression can be dynamically observed in cancer cells in this GFP transgenic mouse model of HCC.