利用萤光素酶标记的肿瘤细胞建立肝癌原位移植模型及其活体成像

Establishment and Bioluminescence Imagining of an Orthotopic Liver Tumor Model Expressing Luciferase

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 investigate the bioluminescent signals of orthotopic liver tumors in nude mice for noninvasive monitoring on progression of liver tumor in vivo. Methods： Human hepatocellular carcinoma cell line HepG2 cells were transduced with vector pCl-neo-Luc encoding luciferase gene. The expression of luciferase in HepG2 cells was analyzed with FACS and bioluminescence imagining. An orthotopic liver tumor model was created with cells from the heterotopic tu- mor established by implanting constitutively tumor cells expressing luciferase subcutaneously into nude mice. The tumor in vivo was monitored by whole-body images and quantity of photon emission. Results： In vitro result shows that photon e- mission correlated with increasing cell numbers. In vivo result indicates the establishment of an orthotopic liver tumor model expressing luciferase. Conclusion： This study provides a platform for monitoring the tumor by orthotopic implantation and evaluating in vivo efficacy of anti-tumor drugs.