生物工程學報 25 August 2009, 25(8):1204～1210

用于活體成像的小鼠肺癌移植瘤模型的建立

張海燕1, 2, 李艷1, 王喆1, 張必良1, 2

1 中國科學院廣州生物醫(yī)藥與健康研究院 呼吸疾病國家重點實驗室, 廣州 510663

2 中國科學技術大學, 合肥 230026

摘  要: 本研究旨在建立可用于活體成像的小鼠肺癌移植瘤模型,。利用脂質體將熒光素酶表達載體pGL4.17(luc2/neo)轉染至人非小細胞肺癌細胞株A549, 經G418篩選獲得穩(wěn)定表達熒光素酶的細胞克隆。根據體外生物發(fā)光情況及細胞的生長特性, 從中挑選合適克隆, 進行裸鼠皮下接種, SCID鼠尾靜脈接種, 建立肺癌移植瘤模型,。利用活體成像系統(tǒng)監(jiān)測腫瘤的生長轉移情況, 并用切片HE染色進一步驗證小鼠模型移植瘤的原位成瘤和轉移能力,。實驗結果表明: 本研究成功地構建了可用于活體成像的小鼠肺癌移植瘤模型, 模型穩(wěn)定可靠、直觀,、靈敏, 為腫瘤生長轉移機制的研究及抗腫瘤藥物的研發(fā)提供了重要工具,。

關鍵詞: 熒光素酶, A549細胞, 動物模型, 移植瘤, 活體成像

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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