小鼠肝癌细胞肺高转移株的建立及鉴定  

Establishment and preliminary identification of a mouse hepatocellular carcinoma cell line with high metastatic potential to the lung

作　　者：王士勇[1,2] 张璐[1] 张晖[1] 杜微丽[1] 张哲[1] 王佳玲[1] 武秀艳[1] 何英[1] 曾雪[1] 刘畅[2] 焦雪[2] 

机构地区：[1]中国医科大学附属第四医院生物治疗科,辽宁省沈阳市110032 [2]中国医科大学附属第四医院生物治疗实验室,辽宁省沈阳市110032

出　　处：《世界华人消化杂志》2011年第16期1674-1679,共6页World Chinese Journal of Digestology

基　　金：教育部归国博士启动课题基金资助项目;No.2004-527~~

摘　　要：目的:建立小鼠肝癌细胞肺高转移株,为研究转移相关分子机制提供合适的模型.方法:将H22肝癌腹水瘤细胞(简称M0)经尾静脉注入昆明小鼠,获取肺转移结节,再制备腹水瘤、尾静脉注射、形成肺转移瘤.重复操作获得第4代肺高转移株(简称M4),检测其小鼠肺转移能力和体外增殖能力、细胞分裂指数、染色体形态及细胞周期比例,并用RT-PCR方法测定RhoC基因mRNA的表达来进一步加以验证.结果:经尾静脉注射,M4较早出现肺转移,肺转移结节数多、体积大.M4与M0体外结果比较:倍增时间缩短了38.73%;细胞分裂指数是4.11%±0.11%,明显多于M0的4.70%±0.19%(P=0.014);S期比例是56.10±4.76%,明显高于M0的46.98±4.49%(P=0.022);二者染色体数目相同,但M4异型性明显;RhoC基因表达分别是1.011±0.163和0.486±0.045,M4的表达显著增高(P=0.0029).结论:建立了小鼠肝癌细胞肺高转移细胞株,其RhoC基因表达明显上调.AIM： To establish a mouse H22 hepatocellular carcinoma cell line with high metastatic potential to the lung to provide a suitable model for the study of metastasis-related molecular mechanisms.METHODS： H22 hepatocellular ascitic tumor cells （M0） were inoculated into mice via the vena caudalis, and pulmonary metastatic lesions were harvested to refabricate ascitic tumor cells. The obtained cells were inoculated into mice again via the vena caudalis to form pulmonary metastatic nodes. The same procedure was repeated four times to obtain a cell line with high metastatic potential to the lung （M4）. The metastatic ability in vivo, proliferation capability in vitro, cell division index, and cell cycle distribution of M4 cells were measured. The mRNA expression of the RhoC gene in M4 cells was detected by RT-PCR. RESULTS： After injection via the vena caudalis, M4 cells produced pulmonary metastasis earlier and formed more and larger nodes. Compared to M0 cells, the doubling time of M4 cells was shortened by 38.73%; cell division index signif icantly increased （P = 0.014）; and the proportion of cells in S phase was significantly higher （P = 0.022）. The number of chromatosomes was comparable between M0 and M4 cells, while heteromorphism was more obvious in M4 cells. The mRNA expression of the RhoC gene was signif icantly higher in M4 cells than in M0 cells （1.011 ± 0.163 vs 0.486 ± 0.045, P = 0.0029）. CONCLUSION： A mouse hepatocellular carci- noma cell line with high metastatic potential to the lung has been established successfully.

关 键 词：肝癌 肺转移 小鼠 细胞株 RHOC 

分 类 号：R735.7[医药卫生—肿瘤]