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快速大容量尾静脉注射法构建循环肝癌细胞肝转移小鼠模型
引用本文:时志龙,孙斌,陈磊,钱海华,张孝峰,殷正丰.快速大容量尾静脉注射法构建循环肝癌细胞肝转移小鼠模型[J].中国肿瘤生物治疗杂志,2014,21(5):574-577.
作者姓名:时志龙  孙斌  陈磊  钱海华  张孝峰  殷正丰
作者单位:苏州大学 医学部 临床医学院,江苏 苏州 215006; 第二军医大学 东方肝胆外科医院 分子肿瘤研究室,上海 200438;第二军医大学 东方肝胆外科医院 分子肿瘤研究室,上海 200438;第二军医大学 东方肝胆外科医院 分子肿瘤研究室,上海 200438;第二军医大学 东方肝胆外科医院 分子肿瘤研究室,上海 200438;第二军医大学 东方肝胆外科医院 分子肿瘤研究室,上海 200438;苏州大学 医学部 临床医学院,江苏 苏州 215006; 2. 第二军医大学 东方肝胆外科医院 分子肿瘤研究室,上海 200438
基金项目:国家传染病重大专项课题资助项目(No.2012ZX10002012-10);国家自然科学基金资助项目 (No.81272669, No.81301830)
摘    要:目的: 通过快速大容量尾静脉注射方法构建循环肝癌细胞肝内复发转移小鼠模型。 方法: 40只C57BL/6J小鼠采用随机数字表法随机分为对照组和实验组(20只/组),对照组采用传统的缓慢小容量尾静脉注射法(2×10 6个肝癌Hepa1-6细胞/0.2 ml,30 s内注射完毕),实验组采用改良的快速大容量尾静脉注射法(2×10 6个肝癌Hepa1-6细胞/2 ml,5 s内注射完毕)构建循环肝癌细胞肝内复发转移小鼠模型。4周后摘取肝、肺、肾、脾组织并行H-E染色,大体和镜下观察肝脏、肺脏、脾脏和肾脏的成瘤情况。 结果: 对照组小鼠只在肺脏有转移灶形成,成瘤率为95%(19/20),肝脏、肾脏、脾脏未见有转移灶形成。实验组小鼠在肝脏和肺脏同时形成转移灶,肺脏成瘤率为94.7%(18/19),肝脏成瘤率为100%(19/19),脾脏、肾脏未见转移灶形成。 结论: 快速大容量尾静脉注射法注射循环肝癌细胞构建的小鼠模型可以模拟肝癌根治性切除后残留的循环肝癌细胞导致早期肝癌肝内复发转移的过程。

关 键 词:循环肝癌细胞  Hepa1-6细胞  肝转移  快速大容量尾静脉注射  动物模型
收稿时间:2014/4/25 0:00:00
修稿时间:2014/8/30 0:00:00

Establishment of a mouse model of hematogenous dissemination of circulating hepatocellular carcinoma cells transplanted via rapid and vast tail vein injection
Shi Zhilong,Sun Bin,Chen Lei,Qian Haihu,Zhang Xiaofeng and Yin Zhengfeng.Establishment of a mouse model of hematogenous dissemination of circulating hepatocellular carcinoma cells transplanted via rapid and vast tail vein injection[J].Chinese Journal of Cancer Biotherapy,2014,21(5):574-577.
Authors:Shi Zhilong  Sun Bin  Chen Lei  Qian Haihu  Zhang Xiaofeng and Yin Zhengfeng
Institution:School of Clinical Medicine, Faculty of Medicine, Soochow University, Suzhou 215006, Jiangsu, China; Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Millitary Medical University, Shanghai 200438, China;Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Millitary Medical University, Shanghai 200438, China;Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Millitary Medical University, Shanghai 200438, China;Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Millitary Medical University, Shanghai 200438, China;Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Millitary Medical University, Shanghai 200438, China;School of Clinical Medicine, Faculty of Medicine, Soochow University, Suzhou 215006, Jiangsu, China; Molecular Oncology Laboratory, Eastern Hepatobiliary Surgery Hospital, Second Millitary Medical University, Shanghai 200438, China
Abstract:Objective : To establish a mouse model of intrahepatic recurrence and metastasis of circulating hepatocellular carcinoma cells transplanted via rapid and vast tail vein injection. Methods: C57BL/6J mice were randomly divided into a control group ( n =20) and an experimental group ( n =20). Animals in the control group were given 2×10 6 human hepatoma Hepa1-6 cells/0.2 ml through conventional speed (30 s) tail vein injection and those in the experimental group were given 2×10 6 Hepa1-6 cells/0.2 ml through rapid (5 s) and vast tail vein injection. Four weeks later, animals were sacrificed, liver, lung, kidney and spleen were collected for H-E staining, and tumor formation was evaluated grossly and microscopically. Results: In the control group, tumor nodules were seen in 19 (95%) mice and the metastatic lesions occurred only in the lung but not in liver, spleen and kidneys. In the experimental group, tumor nodules were present in the lungs in 18 (94.7%) mice and in the liver in 19 (100%) mice while no metastatic tumors were found in the spleen and kidneys. Conclusion: Rapid and vast tail vein injection of hepatoma carcinoma cells in to nude mice may mimic the process of hematogenous dissemination of the residual circulating hepatocellular carcinoma cells after curative resection, thus offering a potentially useful, simple, efficient and safe animal model to study the metastasis of the circulating residential hepatoma carcinoma cells after resection.
Keywords:circulating hepatocellar carcinoma  cell  Hepa1-6  liver metastases  rapid and vast tail vein injection  animal model
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