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目的:利用生物发光成像技术非侵入性地监测活体裸鼠原位肝癌发展过程。方法:将包含有萤火虫萤光素酶基因的pCI-neo-Luc载体转染人肝癌HepG2细胞系,筛选获得具有高萤光素酶活性的细胞克隆;利用流式细胞仪对萤光素酶表达的稳定性进行初步研究,并分析细胞的生物发光情况;持续表达萤光素酶的肿瘤细胞培养扩增后被植入裸鼠皮下,2周后以形成的异体瘤作为供体瘤,进行肝脏原位移植手术;对建立的肝癌原位移植模型,用影像学资料显示肿瘤部位,用IVIS成像系统动态监测肿瘤生长情况。结果:体外影像的结果显示,表达萤光素酶细胞的数量与发光强度呈正相关;活体成像的结果显示,成功地建立了萤光素酶标记的原位肝癌动物模型。结论:生物发光成像可以监测活体内肝癌演进过程,为抗肿瘤药物的筛选和评价提供了新的手段和工具。
Objective: To monitor non-invasively the development of orthotopic liver cancer in nude mice using bioluminescence imaging technology. METHODS: Human hepatoma HepG2 cell line was transfected with pCI-neo-Luc vector containing firefly luciferase gene, cell clones with high luciferase activity were screened, and luciferase expression was stabilized by flow cytometry. A preliminary study was conducted and the bioluminescence of the cells was analyzed. The tumor cells that continuously expressed luciferase were cultured and expanded and implanted subcutaneously in nude mice. After 2 weeks, the formed allografts were used as donor tumors to perform the liver in situ. Transplantation; Established models of orthotopic transplantation of hepatocellular carcinoma, using imaging data to show tumor sites, and using IVIS imaging system to dynamically monitor tumor growth. Results: The results of in vitro imaging showed that the number of luciferase-expressing cells was positively correlated with luminescence intensity; the results of in vivo imaging showed that a luciferase-labeled orthotopic liver cancer animal model was successfully established. Conclusion: Bioluminescence imaging can monitor the evolution of liver cancer in vivo and provide new tools and tools for the screening and evaluation of anti-tumor drugs.