论文部分内容阅读
目的:建立一株新的肝癌细胞系EHBC-512并分析其生物学特性,为后续研究奠定基础。方法:切取肝癌手术标本进行体外原代培养,采用光镜、电镜观察细胞形态,流式细胞检测细胞周期,并进行染色体核型分析,免疫荧光及化学发光法检测细胞上清液AFP的表达;裸鼠体内种植观察异种成瘤情况。结果:光镜下细胞形态具有肝癌细胞特征;电镜下细胞具有较多的线粒体,核异形性明显,染色体众数为110～120条。细胞群体倍增时间为48h,贴壁率达90%。免疫荧光染色显示:细胞AFP、CK18表达,CEA、CK19未表达;细胞高分泌AFP,上清液AFP含量>1210μg/L。裸鼠体内成瘤病理检测与原发灶保持相同的结构和细胞形态。结论:成功建立一株生物学特性稳定并能体外高分泌AFP的人类肝癌细胞系(EHBC-512),为肝癌研究提供新的研究工具。
OBJECTIVE: To establish a new hepatocellular carcinoma cell line EHBC-512 and analyze its biological characteristics, and lay the foundation for further research. Methods: Primary hepatocellular carcinoma (HCC) specimens were excised for primary culture in vitro. Morphological changes were observed under light and electron microscopes. Cell cycle was detected by flow cytometry. Chromosome karyotype analysis was performed. The expression of AFP was detected by immunofluorescence and chemiluminescence. Nude mice were seeded in vivo tumorigenesis. Results: The morphological features of hepatocellular carcinoma cells under light microscope showed that the cells had more mitochondria under electron microscope, and the shape of nucleus was obvious. The number of chromosomes was 110-120. Cell population doubling time 48h, adherent rate of 90%. Immunofluorescence staining showed that the expression of AFP and CK18 was not expressed in CEA and CK19; AFP was highly secreted by cells and the AFP content in supernatant was> 1210 μg / L. Histopathology of nude mice in vivo detection and primary tumor to maintain the same structure and cell morphology. Conclusion: A human hepatocellular carcinoma cell line (EHBC-512) with stable biological characteristics and high secretion of AFP in vitro was successfully established, which provided a new research tool for the study of liver cancer.