生物工程學(xué)報(bào) 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(guó)科學(xué)院廣州生物醫(yī)藥與健康研究院 呼吸疾病國(guó)家重點(diǎn)實(shí)驗(yàn)室, 廣州 510663
2 中國(guó)科學(xué)技術(shù)大學(xué), 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(zhì)體將熒光素酶表達(dá)載體pGL4.17(luc2/neo)轉(zhuǎn)染至人非小細(xì)胞肺癌細(xì)胞株A549, 經(jīng)G418篩選獲得穩(wěn)定表達(dá)熒光素酶的細(xì)胞克隆,。根據(jù)體外生物發(fā)光情況及細(xì)胞的生長(zhǎng)特性, 從中挑選合適克隆, 進(jìn)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(cè)腫瘤的生長(zhǎng)轉(zhuǎn)移情況, 并用切片HE染色進(jìn)一步驗(yàn)證小鼠模型移植瘤的原位成瘤和轉(zhuǎn)移能力。實(shí)驗(yàn)結(jié)果表明: 本研究成功地構(gòu)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(zhǎng)轉(zhuǎn)移機(jī)制的研究及抗腫瘤藥物的研發(fā)提供了重要工具。
關(guān)鍵詞: 熒光素酶, A549細(xì)胞, 動(dòng)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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