犬乳腺肿瘤裸鼠模型建立及其生长特性

Establishment of a Nude Mouse Model of Canine Mammary Tumor and Its Growth Characteristics

为研究犬乳腺肿瘤发展的生物学特性及动态演变过程,建立犬乳腺肿瘤裸鼠模型,探讨其发生及发展规律。将绿色荧光蛋白(GFP)导入到犬乳腺肿瘤CHMm系和CHMp系细胞,将细胞悬液注射种植至裸鼠右侧第二对乳头处,建立裸鼠模型。通过活体成像技术与病理学检测技术,研究不同时间段裸鼠体内肿瘤的动态变化。结果显示:倒置荧光显微镜下可观察到GFP标记的CHMm-GFP和CHMp-GFP细胞均发出稳定的绿色荧光信号,种植后第6天出现肉眼可见肿瘤。通过对接种后第25、35、45和60天荧光信号进行检测和病理检查,绘制肿瘤生长曲线和体重变化曲线;且在接种后第25天裸鼠活体成像时,CHMm-GFP系裸鼠体内荧光信号最强,而CHMp-GFP系则在第35天时,裸鼠体内荧光信号最强。在接种后第25天的离体脏器成像检测时,发现两组裸鼠各有1只淋巴结开始显现荧光信号;第35天3只裸鼠淋巴结内均显示荧光信号,其他脏器无荧光信号。第25天病理组织切片检测,两种细胞系分别有1只裸鼠的肿瘤转移至淋巴结内;在第35、45和60天时解剖裸鼠,发现肿瘤均已转移到淋巴结。建立了两个细胞系犬乳腺肿瘤裸鼠模型,两个细胞系在裸鼠体内不同时段呈现不同的生长特性,且均可转移,为深入研究乳腺肿瘤发生和发展规律提供参考。

This research was conducted to establish a nude mice model of canine mammary cancer cell,and study its biological characteristics as well as dynamic changes.The green fluorescent protein(GFP)was introduced into the CHMm and CHMp cells of canine breast cancer cell.The nude mice model was established by implantation of tumor cell suspension in right second breast of the nude mice.The dynamic changes of tumor in nude mice at different time periods were studied by the technique of living fluorescence imaging and pathological examination.Results were as follows:GFP labeled CHMm-GFP and CHMp-GFP cells were observed to emit stable green fluorescence signals under inverted fluorescence microscope,and macroscopic tumor appeared on the 6th day after implantation.The tumor growth curve and weight change curve were drawn by fluorescence signal detection and pathological examination on the 25th day and the 60th day after implantation.On the 25th day after implantation,the fluorescence signal of CHMm-GFP was the strongest in nude mice,while that of CHMp-GFP was strongest on 35th day.On the 25th day after implantation,it was found that one lymph node of each group of nude mice showed fluorescence signal.On the 35th day,fluorescent signals were displayed in lymph nodes of 3 nude mice,but no fluorescence signals were found in other organs.On the 25th day,one nude mouse tumor metastasized to the lymph node,and at the 35th day and the 60th day,the tumor was found to have metastasized to the lymph node.Canine breast tumor models of two cell lines were established in this study.The two cell lines showed different growth characteristics in nude mice at different periods and could be metastasized,which provided a reference for the further study of the occurrence and development of mammary tumors.