SPC-CRE-Kras双阳性转基因小鼠自发肺部肿瘤模型的建立  

The establishment of SPC-CRE-Kras double transgenic spontaneous pulmonary tumor mouse model

作　　者：高昆[1] 刘学丽[1] 高珊[1] 高凯[1] 董伟[1] 张旭[1] 刘宁[1] 徐艳峰[1] 张连峰[1] 

机构地区：[1]中国医学科学院,北京协和医学院医学实验动物研究所,卫生部人类疾病比较医学重点实验室,北京100021

出　　处：《中国比较医学杂志》2013年第7期11-15,I0001,共6页Chinese Journal of Comparative Medicine

基　　金：国家科技支撑计划课题(2012BA139B02);国家“重大新药创制”科技重大专项课题(课题编号2011ZX09307-302)

摘　　要：目的构建一种利用Cre重组酶体内低表达诱导K-ras G12D在小鼠肺部活化的慢性自发性肺部肿瘤模型。方法首先构建一种肺脏特异性低表达Cre重组酶的SPC-CRE转基因小鼠,利用SPC-CRE转基因小鼠与LSL K-ras G12D转基因小鼠杂交,获得SPC-CRE-Kras双阳性转基因小鼠。对4月龄,5月龄,7月龄,9月龄SPC-CRE-Kras双阳性转基因小鼠肺部进行取材,固定并进行常规石蜡切片及HE染色,镜下观察小鼠肺部病理特征。使用micro-CT对7月龄,9月龄SPC-CRE-Kras双阳性转基因小鼠肺部肿瘤结节进行检测。结果获得了SPC-CRE-Kras双阳性转基因小鼠,该小鼠在肺组织特异性低表达Cre重组酶,并诱导K-ras G12D在肺组织的表达,由K-ras G12D引起肺组织肿瘤的发生。SPC-CRE-Kras双阳性转基因小鼠4月龄肺部出现轻度炎症反应,5月龄开始肺部可见散在腺瘤样的结节,成瘤率为100%(雌6/6,雄6/6),随着月龄增加,小鼠肺腺瘤结节呈增大趋势,病理变化呈进展状态,9月龄时通过micro-CT可以检测到肺部少量散在孤立的肿瘤结节。结论利用肺组织特异性低表达Cre重组酶的方式,建立了一种从肺部炎症反应到肺腺瘤进展时程较长的慢性自发肺部肿瘤小鼠模型,为肺癌发生的研究提供了更长的窗口期。Objective To establish a spontaneous pulmonary tumor mouse model in which the K-Ras gene was activated by Cre/loxp recombinant enzyme system. Methods SPC-CRE transgenic mice were generated that lowly express lung-specific Crc recombinant enzyme. The SPC-CRE transgenic mice were mated with LSL K-ras G12D transgenic mice to produce SPC-CRE-Kras double transgenic mice. The 4, 5, 7 and 9 month-old SPC-CRE-Kras double transgenic mice were sacrificed and the lung tissues were extracted, fixed, embedded in paraffin and sliced. Hematoxylin-eosin （HE） staining was performed and observed under light microscope. Micro-CT was used to test the pulmonary nodules of SPC-CRE-Kras double transgenie mice. Results The SPC-CRE-Kras double transgenic mice were generated. The expression of K-ras G12D in the SPC-CRE-Kras double transgenic mice could be induced by the Cre/Loxp recombinant enzyme system. Mild pulmonary inflammation could be found in the 4 month-old SPC-CRE-Kras double transgenic mice. The sporadic adenoma could be found in the lung of 5 month-old mice, with 100% of the mice developing pulmonary adenoma （ female 6/6, male 6/6）. The size and progression of the adenoma is time dependent. The pulmonary nodules could be determined by micro- CT in the 9 month-old. Conclusions A chronic spontaneous pulmonary tumor mouse model was established by hybridization. Longer progressive period from inflammation to adenoma in this model will provide enough time for the investigation of pulmonary tumorigenesis.

关 键 词：CRE Loxp重组酶系统 转基因小鼠 K-RAS基因 小鼠肿瘤模型 

分 类 号：R-332[医药卫生]