| 摘要: |
| 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48 h，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>1 210 μg/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。 |
| 关键词: 肝肿瘤 肝细胞癌 细胞培养 细胞系 生物学特性 |
| DOI：10.3724/SP.J.1008.2009.01115 |
| 投稿时间：2009-03-01修订日期：2009-09-04 |
| 基金项目:艾滋病和病毒性肝炎等重大传染病防治专项基金(2008ZX10002-025). |
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| Establishment and characterization of a human hepatocellular carcinoma cell line EHBC-512 |
| ZOU Qi-fei1,HOU Ying2,LIU Jian1,GE Rui-liang1,SHEN Feng1* |
| (1.Department of Comprehensive Treatment Ⅰ,Eastern Hepatobiliary Hospital,Second Military Medical University,Shanghai 200438,China;2.Institute of Neuronal Science,Chinese Academy of Science,Shanghai 200031) |
| Abstract: |
| Objective:To establish a new hepatocellular carcinoma cell line EHBC-512 and to analyze its biological property,so as to lay a foundation for future study.Methods:The specimen of liver cancer was resected during operation and was primary cultured.The morphology of cells was observed under optical and electron microscope.The cell cycle was examined with flow cytometry; the chromosome karyotype was also analyzed.The expression of AFP in the culture supernatant was detected by immunofluorescence and chemoluminescence.The cultured cells were implanted into nude mice for observation of tumor formation.Results:The cultured cell line had the character of liver cancer under light microscope,and had rich mitochondria,with obvious nuclear heteromorphism and a chromosome number of 110-120.The doubling time of the cells was 48 h,and the adherence rate was 90%.Immunofluorescence showed that the cells were positive of AFP and CK18 and negative of CEA and CK19.The production of AFP was higher and the content of AFP was higher than 1 210 μg/L.The histology of the exografts was similar to that of the primary tumor.Conclusion:We have successfully established a new hepatocellular cell line EHBC-512,which can stably secrete AFP in vitro,providing a novel tool for liver cancer research. |
| Key words: liver neoplasms hepatocellular carcinoma cell culture cell line biological characterization |
