人肝癌细胞系HCC-9724的建立及其生物学特性

Establishment of a human hepatocyte carcinoma cell line HCC-9724 and its biological properties

目的 建立一株肝癌细胞系 HCC- 972 4,为肝癌研究提供实验模型 .方法 无菌切除肝癌手术标本 ,6只裸鼠右后肢皮下包埋 ,体内连续传 3代后 ,取移植瘤体外培养 ,绘制细胞生长曲线 ,光镜、电镜观察细胞形态 ,并进行细胞周期和染色体核形分析 ,用免疫荧光法测定荷瘤鼠中 AFP的表达 .结果 移植模型原代获得率为 33.3% ,潜伏期为 5 2 d,自第 3代后肿瘤获得率为 10 0 .0 % ,潜伏期为 16 d,该模型移植瘤具有生长稳定 ,移植成功率高 (10 0 % ) ,潜伏期短 (10 d) ,无自发消退等特点 ,裸鼠移植瘤标本体外培养得到肝癌细胞系 HCC-972 4(简称 H) ,其形态结构 ,排列方式和移植瘤的分化程度与原发瘤保持一致 ,且遗传学特性包括染色体形态仍为人类核型 ,众数维持在 6 2～ 6 6之间 ,占 75 .0 % ,细胞周期分析 G1 期5 3.5 % ,G2 期 19.5 % ,S1 期 2 7.0 % ,细胞群体倍增时间为 2 3d,建立的裸小鼠人肝癌模型具有典型的肝细胞癌特征 ,持续分泌甲胎蛋白 (AFP) .结论 通过裸鼠体内移植瘤建立的肝癌细胞系 HCC- 972 4,与原发癌保持相同的生物学特性 ,体外连续传代 6 mo以上 ,细胞形态不变 ,生长周期恒定 。

AIM To establish a stain of hepatocyte carcinoma cell line and to provide hepatic carcinoma animal model. METHODS The liver tumor specimens incised sterilely were inoculated subcutaneously into nude mice, and the third generation tumor tissue was cultured in vitro . A tumor cellgrowth curve was drawn. Morphology of the cells was observed. Cell cycling analysis and chromosome analysis were performed. AFP expressed on live carcinoma nude mice was tested by immunofluorescence technique. RESULTS In the primary inoculation the rate was 33.3%; the latent period was 52 days. In No. 3 passage, the rate was 100%; the latent period was 16 days. We incised the tumor mass from the nude mouse cultured, and obtained a hepatic tumor cell line. Its structure, arrangement and differentiation remained unchanged. In addition, its chromosomal structure was still like humans. Mode rage was 72～76 (75%).The cycling analysis showed 53.2% G 1,17.0% G 2 and 29.8% S 1. The cell had a doubling time of 23.0 hours. Nude mice tumor model could secrete AFP continuously. CONCLUSION The hepatocyte carcinoma cell line HCC 9724 established by tumor subcutaneous transplanting in nude mice was identified with primary cancer cell in biological characters. Passaging continually for more than half a year, its characteristics were still maintained. Therefore, HCC 9724 may be used as a stable cell line.