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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 ,可望作为一个稳定的细胞系
Objective To establish a hepatocellular carcinoma cell line HCC-9724. To provide an experimental model for the study of hepatocellular carcinoma. METHODS: Hepatoma surgical specimens were aseptically resected. Six nude mice were implanted subcutaneously in the right hind limb. After three consecutive passages in vivo, the xenografts were cultured in vitro. The cell growth curve was drawn, the cell morphology was observed by light and electron microscopy, and the cell cycle and chromosome karyotype analysis were performed. The expression of AFP in the tumor-bearing mice was measured by immunofluorescence. The results showed that the primary acquisition rate of the transplanted model was 33.3% and the incubation period was 5 After 2 days, the tumor acquisition rate was 100.0% and the latency was 16 days after the third passage. The transplanted tumor in this model had stable growth, high success rate (100%), short incubation period (10 days), no spontaneous Regression and other characteristics, nude mice transplanted tumor specimens in vitro cultured to obtain hepatocellular carcinoma cell line HCC-972 4 (abbreviation H), its morphological structure, arrangement and the degree of differentiation of transplanted tumors consistent with the primary tumor, and genetic characteristics including chromosome morphology The human karyotype is still maintained, the mode is maintained between 6 2 and 6 6 , accounting for 75.0 %, cell cycle analysis G1 phase 5 3.5 %, G2 phase 19.5 %, S1 phase 2 7.0 %, cell population doubling time is 2 3d, Established nude mouse human liver cancer The type has a typical hepatocellular carcinoma characteristic and continuously secretes alpha-fetoprotein (AFP). Conclusion The hepatocellular carcinoma cell line HCC-9724, which was established by transplantation tumor in nude mice, maintains the same biological characteristics as the primary cancer, and is serially passaged in vitro. More than mo, the cell morphology is unchanged, the growth cycle is constant, and it is expected to be a stable cell line.