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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.
OBJECTIVE: To establish a mouse model of lung metastasis of hepatocellular carcinoma in mice and to provide a suitable model for studying the molecular mechanism of metastasis.Methods: H22 hepatocellular carcinoma ascites tumor cells (M0) were injected into Kunming mice through tail vein to obtain lung metastasis nodules Preparation of ascites tumor, tail vein injection, the formation of lung metastases. Repeated operation to obtain the fourth generation of lung metastasis (referred to as M4), lung metastasis ability and in vitro proliferation ability, cell division index, chromosome morphology and cell cycle ratio , And the expression of RhoC gene mRNA was further confirmed by RT-PCR method.Results: M4 was earlier appeared lung metastasis, pulmonary metastasis nodules were more and larger volume by tail vein injection.M4 and M0 in vitro results were compared: doubling time (P = 0.014), the percentage of S phase was 56.10 ± 4.76%, which was significantly higher than that of M0 46.98 ± 4.49% (P = 0.014) P = 0.022). The chromosome number of the two was the same, but the M4 atypia was obvious. The expression of RhoC gene was 1.011 ± 0.163 and 0.486 ± 0.045 respectively, and the expression of M4 was significantly increased (P = 0.0029) .Conclusion: Highly metastatic cell lines, RhoC gene expression was significantly up-regulated.