用于活体成像的人肺腺癌自发骨转移小鼠模型  

Establishment of spontaneous bone metastasis mouse models of lung adenocarcinoma for in vivo imaging system

作　　者：韩帅[1] 杨兴海[1] 万宗淼[1] 周旺[1] 姜东杰[1] 尹华斌[1] 王霆[1] 肖建如[1] 

机构地区：[1]上海长征医院骨肿瘤科,200003

出　　处：《国际骨科学杂志》2015年第2期151-156,共6页International Journal of Orthopaedics

基　　金：上海市科委实验动物专项基金(12140901900);上海市卫生系统优秀青年人才培养计划(XYQ2013099)

摘　　要：目的建立小鼠人肺腺癌PC-9细胞株骨转移模型,初步探讨由模型动物中分离上皮-间质细胞转化(EMT)癌细胞的可行性。方法采用肺原位注射法对低、中、高浓度组的雄性BALB/c裸鼠分别注射1×104、5×104、1×105个/20μL的高骨亲和性的人肺腺癌PC-9细胞悬液,对照组注射5×104/20μL的原始人肺腺癌PC-9细胞,空白组注射20μL空白混合胶体,观察小鼠的生存时间、体重变化及肺腺癌骨转移模型的成模率,并采用流式细胞技术对分离外周血EMT癌细胞进行鉴定。结果注射PC-9细胞后21 d,对照组和低、中、高浓度组小鼠均出现体重下降、摄食减少等,与空白组相比体重明显降低(P<0.05)。高浓度组小鼠注射PC细胞后20 d出现骨转移病灶,27 d出现死亡。低、中、高浓度组的肺腺癌骨转移模型成模率分别为0%(0/5)、25%(2/8)、60%(6/10)。造模成功的小鼠肺部和骨骼出现转移灶,病理检查可见肺腺癌细胞,其外周血可成功分离EMT癌细胞,其中EMT癌细胞阳性小鼠骨转移发生率较高。结论本研究成功构建了可用于活体成像的人肺腺癌骨转移小鼠模型,并可从其外周血中分离EMT癌细胞,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。Objective To establish mouse models of lung adenocarcinoma spontaneous bone metastasis and perform a preliminary study on isolating the epithelial-mesenchymal transition（EMT） cells from the mouse models. Methods We injected different concentration bone-seeking PC-9 cells with Luciferase and GFP gene expression into the lung of the BALB/c nude mice. The survival time, bodyweight, tumor formation rate in the lung and metastasis rate to other organs were observed. Then we analyzed the EMT cells of the mouse models by in vitro bioluminescence imaging and flow cytometry analysis. Results Compared with the normal group, the bodyweight and daily food intake of the mouse-model groups decreased from day 21 after the injection of tumor cells. Bone metastasis occurred on day 20 and death on day 27 after the injection of tumor cells in high-concentration of the high-concentration, mid-concentration and low-concentration group were 60 oup. The bone metastasis rate （6/10）, 25%（2/8） and 0% （0/5） respectively, which were higher in the EMT cells positive mice. Conclusion Spontaneous bone metastasis mouse models of human lung adenocarcinoma for in vivo imaging were successfully established, from which EMT cells can be isolated. Such models could be used for studies on the mechanisms of lung cancer and the development of anticancer drugs.

关 键 词：动物模型 PC-9细胞 活体成像 肺腺癌骨转移 上皮-间质转化 

分 类 号：R734.2[医药卫生—肿瘤] R-332[医药卫生—临床医学]