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本研究应用人肝癌细胞SCID鼠转移模型,筛选和建立肝癌转移细胞系,旨在为肝癌转移研究提供实验材料.采用人肝癌细胞H7402,腋后背部皮下接种SCID鼠,接种后66天时,在荷瘤鼠赢弱时引颈处死,取其肺组织,进行原代细胞培养和传代培养,获得肝癌细胞H7402的亚细胞克隆,命名为M-H7402.然后,对M-H7402细胞进行了生物学鉴定,检测了细胞形态、染色体、细胞动力学、细胞周期、甲胎蛋白表达、癌基因表达和转移相关基因表达等指标.结果显示,M-H7402细胞的生长、增殖和形态等与亲本细胞H7402十分相近,其染色体形态仍为人类核型,众数维持在72~80之间,占75.0％.RT-PCR检测结果显示,M-H7402细胞甲胎蛋白表达为阳性。Western blot检测结果显示,与亲本细胞H-7402相比,癌基因C-myc表达水平较高,转移抑制基因nm23的表达水平明显下调.从肺组织中获得的肝癌转移细胞系M-H7402,在体外可连续传代培养,细胞形态不变.应用SCID鼠筛选获得的亚细胞克隆,与亲本细胞形成配对关系,可为肝癌细胞转移的研究提供新的实验材料.
In this study, human hepatocellular carcinoma cell line SCID was used to screen and establish a metastatic cell line of hepatocellular carcinoma (HCC), and to provide experimental materials for HCC metastasis.Using H7402 human hepatocellular carcinoma cells, SCID mice were subcutaneously inoculated on the back of the axillary gyrus and 66 days after inoculation Mice were sacrificed and their necks were sacrificed. The lungs were harvested for primary culture and subculture. The subcellular clones of H7402 cells were obtained and named as M-H7402.M-H7402 cells were identified by their biological characteristics, The cell morphology, chromosome, cell kinetics, cell cycle, alpha-fetoprotein expression, oncogene expression and metastasis-related gene expression were detected.The results showed that the growth, proliferation and morphology of M-H7402 cells were very similar to the parental cells H7402 , The chromosome shape is still human karyotype, the number maintained at 72 to 80, accounting for 75.0% .RT-PCR test results showed that M-H7402 cells were positive for alpha-fetoprotein. The results of Western blot showed that the expression of oncogene C-myc was higher than that of parental H-7402 and the expression of metastasis suppressor gene nm23 was down-regulated.M-H7402, a metastatic hepatoma cell line derived from lung tissue, The cells can be subcultured in vitro and the morphology of the cells remains unchanged.The subcellular clones screened by SCID mice are paired with the parental cells to provide new experimental materials for the study of metastasis of hepatocellular carcinoma.