生物工程学报 25 August 2009, 25(8):1204~1210
用于活体成像的小鼠肺癌移植瘤模型的建立
张海燕1, 2, 李艳1, 王喆1, 张必良1, 2
1 中国科学院广州生物医药与健康研究院 呼吸疾病国家重点实验室, 广州 510663
2 中国科学技术大学, 合肥 230026
摘 要: 本研究旨在建立可用于活体成像的小鼠肺癌移植瘤模型。利用脂质体将荧光素酶表达载体pGL4.17(luc2/neo)转染至人非小细胞肺癌细胞株A549, 经G418筛选获得稳定表达荧光素酶的细胞克隆。根据体外生物发光情况及细胞的生长特性, 从中挑选合适克隆, 进行裸鼠皮下接种, SCID鼠尾静脉接种, 建立肺癌移植瘤模型。利用活体成像系统监测肿瘤的生长转移情况, 并用切片HE染色进一步验证小鼠模型移植瘤的原位成瘤和转移能力。实验结果表明: 本研究成功地构建了可用于活体成像的小鼠肺癌移植瘤模型, 模型稳定可靠、直观、灵敏, 为肿瘤生长转移机制的研究及抗肿瘤药物的研发提供了重要工具。
关键词: 荧光素酶, A549细胞, 动物模型, 移植瘤, 活体成像
Establishment of xenograft mouse models to study human lung cancer by using in vivo imaging system
Haiyan Zhang1, 2, Yan Li1, Zhe Wang1, and Biliang Zhang1, 2
1 State Key Laboratory for Respiratory Diseases, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510663, China
2 University of Science and Technology of China, Hefei 230026, China
Abstract: We established xenograft mouse models for studying human lung cancer by using an in vivo imaging system. We first transfected pGL4.17 (luc2/neo) plasmid into human non-small lung cancer A549 cells and screened cell lines stably expressing a luciferase reporter gene with G418. Then we analyzed the correlation of luciferase activity and cells number by in vitro bioluminescence. Furthermore, we compared cell growth characteristics by cell counting. We selected suitable clones and inoculated subcutaneously into nude mice or intravenously into SCID mice to construct lung cancer xenograft models. Using an in vivo imaging system, we monitored the growth and metastasis of the tumors. Finally, we verified the extents of tumorigenesis and metastasis by tissue sections with Hematoxylin and Eosin (HE) staining. In our study, we successfully established the xenograft mouse models for in vivo imaging with luciferase expressed lung cancer cells. These models provided convenient, sensitive, intuitive and stable tools for studying the mechanisms of lung cancer progression and development of anticancer drug.
Keywords: luciferase, A549 cell, animal model, xenograft, in vivo imaging system
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