人肝细胞癌细胞系HCC-9903的建立及其生物学特性研究

Establishment and biologic characterization of human hepatocellular carcinoma cell line HCC-9903

目的 新建 1株人体肝细胞肝癌细胞系 ,为肝癌研究提供新的、理想的实验模型。 方法 以手术切除肝癌标本作原代培养 ,建立细胞系。通过光镜、电镜、倍增时间、生长曲线、双层软琼脂培养、流式细胞仪、染色体分析、刀豆球蛋白凝集试验、裸鼠接种、AFP、HBV及端粒酶检测 ,对其生物学特性进行分析。 结果 第 2 9天完成原代培养。形态学、增殖动力学及浸润性生长结果表明该细胞系符合恶性细胞特征。染色体数目分布在 49～ 135 ,众数为 6 0～ 6 7,1q(i) 和t(6 ,11)等为其标记染色体 ;有四射体和核内复制出现。裸鼠接种成瘤 ,瘤细胞形态与原病人的病理切片相似。端粒酶检测强阳性。 结论 该细胞系符合建系标准 ,是 1株新建的人体肝细胞肝癌细胞系 ,命名为HCC 990 3。

Objective To establish a new hepatocellular carcinoma (HCC) cell line and provide a new and ideal experimental model. Methods The cell line designated as HCC 9903 was derived from a surgical specimen of HCC. After primary culture, its biologic characteristics were studied under light and electron microscope and by determination of doubling time and growth curve, culture in soft agar, flow cytometry(FCM), chromosome analysis, detection of AFP, HBV and telomerase, agglutination test of concanavalin (ConA) as well as inoculation in nude mice. Results The first passage was found on the 29th day. Morphological study demonstrated its malignant characteristics. Chromosome analysis revealed a chromosome number ranging from 49 to 135, with a modeal number of 52 to 67. Aberrant chromosomes 1q(i) and t (6,11)were its chromosome markers. The cell line produced tumors after subcutaneous transplantation into nude mice. Conclusion According to the rules of establishing a cell line in vitro, HCC 9903 has been proved to be a new cell line of human HCC carcinoma, and more experimental methods have been used to study its biological characteristics.