两种方法构建自发转移且便于观察的裸鼠原位肝癌模型  

Establishing a spontaneous metastasis and conveniently monitored orthotopic model of hepatocellular carcinoma through two methods in nude mice

作　　者：郭平 许敏华 张晶晶[1] 金小宝[1] 李小波[1] 马艳[1] GUO Ping;XU Minhua;ZHANG Jingjing;JIN Xiaobao;LI Xiaobo;MA Yan(Guangdong Key Laboratory of Pharmaceutical Bioactive Substances,Guangdong Pharmaceutical University,Guangzhou 510006,China)

机构地区：[1]广东药科大学生物活性药物研究重点实验室,广州510006

出　　处：《中国比较医学杂志》2020年第3期32-38,共7页Chinese Journal of Comparative Medicine

基　　金：广东省自然科学基金(2016A030310299);广东省科技计划项目(2017A09090544)。

摘　　要：目的构建Luc-GFP标记的高转移性人肝癌细胞株,两种方法建立方便观察且自发转移的裸鼠原位肝癌模型。方法构建p CDH-CMV-Luc-EF1-GFP-Puro重组慢病毒表达载体,并转染人肝癌细胞株HCCLM3,通过细胞悬液肝左叶注射法或皮下移植瘤组织块肝内植入法构建裸鼠原位肝癌模型,利用活体成像技术和组织病理学比较两种方法肿瘤的生长和转移情况。结果构建稳定表达荧光素酶(Luc)和绿色荧光蛋白(GFP)的高转移性人肝癌细胞株HCCLM3-Luc-GFP,裸鼠活体成像和病理学结果发现,两种方法建立的原位肝癌模型成瘤率均为100%。其中细胞悬液法建模周期短,对宿主肝的损伤小,更能模拟出肝癌患者的微环境,转移率高;而肿瘤组织块法的成瘤大小和位置较易控制,但转移率较低。结论成功构建Luc-GFP标记的高转移性人肝癌细胞系HCCLM3-Luc-GFP,细胞悬液法和肿瘤组织块转接法分别建立了自发转移且便于观察的裸鼠原位肝癌模型,为连续、直观地监测肿瘤的生长转移,评价抗肝癌生长转移药物的疗效奠定了基础。Objective To construct a Luc-GFP-labeled human hepatocellular carcinoma( HCC) cell line with high metastatic potential. In addition,to establish a spontaneous metastasis and conveniently monitored orthotopic model of hepatocellular carcinoma through a cell suspension injection method or a tumor fragment implantation method in nude mice.Methods The recombinant lentiviral expression vector p CDH-CMV-Luc-EF1-GFP-Puro was constructed and transfected into the human hepatoma cell line HCCLM3. The orthotopic xenograft model was established through the cell suspension injection method and the tumor fragment implantation method. The growth and metastasis of the tumors were observed by in vivo imaging and pathological analysis. Results HCCLM3-Luc-GFP,a highly metastatic HCC cell line with GFP expression and Luc activity,was obtained. The tumorigenic rates of the two method were 100%. However,the former can simulate the microenvironment of liver cancer patients,with host factors having less influence. The tumor size and location of the latter are easier to control,with the lung metastatic rate being lower in the former than in the latter. Conclusions The Luc-GFP-labeled highly metastatic human hepatoma cell line HCCLM3-Luc-GFP was successfully constructed. The orthotopic model of highly metastatic and Luc-GFP-labeled HCC in nude mice was successfully established by the above method. This study provides a continuous and effective means to monitor the growth and metastasis of tumors in vivo,for evaluating the efficacy of anti-metastatic drugs against HCC.

关 键 词：裸鼠 原位肝癌模型 活体成像 高转移性人肝癌细胞 

分 类 号：R-33[医药卫生]