小鼠H22肝癌肺转移模型的建立及其意义

Establishment of Pulmonary Metastasis Model of Mice H22 Liver Neoplasms and Its Significance

目的:建立小鼠H22肝癌肺转移模型,为后续研究肝癌的侵袭转移机制奠定实验基础。方法:复苏H22肝癌细胞,在培养皿中进行传代培养并计数,当细胞达到预定数目时,收集处于对数期细胞,用生理盐水洗涤2次,调整细胞浓度为1.0×10~7/ml,取0.5ml细胞悬液在无菌环境下注射到小鼠腹腔进行体内传代培养,收集小鼠腹水瘤细胞,用生理盐水洗涤2次,细胞浓度调至2×10~7/ml,取50μl细胞悬液通过小鼠尾静脉注射到小鼠体内,14d后人道主义处死小鼠,解剖取出小鼠全肺组织,病理组织学方法鉴定肿瘤组织,全肺连续切片,显微镜下计数肿瘤转移灶数目。结果:全部小鼠肺组织均肿瘤转移灶形成,肉眼可见成肺组织表面微小转移灶,病理鉴定为肝癌肺转移灶,小鼠肺部肿瘤转移灶数平均为23个。结论:成功建立了小鼠H22肝癌肺转移模型,为研究肝癌转移机制提供了理想的动物模型。

Objective：To establish the model of lung metastasis of H22 hepatocellular carcinoma in mice,and to lay the experimental foundation for the follow-up study on the mechanism of invasion and metastasis of HCC.Methods：H22hepatocarcinoma cells were resuscited and counted in culture dishes.When the cells reached a predetermined number,the cells were collected in logarithmic phase,washed twice with normal saline,adjusted to cell concentration of 1.0×107/ml,0.5ml of cell suspension was injected into the mouse peritoneum in a sterile environment for in vivo subculture.The mice ascites tumor cells were collected,washed twice with physiological saline,adjusted to 2×107/ml and 50μl of cell suspension.The mice were sacrificed in mice,and the mice were sacrificed in 14 days.The whole lung tissue was taken out from the mice.The histological methods were used to identify the tumor tissue and the whole lungs were sliced.The number of tumor metastases was counted under the microscope.Results：All the mice lung tissue tumor metastasis,the naked eye can be seen as the surface of lung tissue micro-metastases,pathological identification is liver cancer lung metastases,the number of lung cancer in each mouse an average of 23 metastases.Conclusion：The model of lung metastasis of H22 hepatocellular carcinoma in mice was established successfully,which provided an ideal animal model for studying the metastasis mechanism of H22 liver cancer.