摘要
目的建立具有器官转移亲嗜性的单克隆人肝癌细胞株和相应的裸鼠移植模型。方法取肺和淋巴结转移亲嗜性人肝癌荧光细胞株HCCLM3-R-LMI及HCCLM3-R-LnM1，通过极限稀释法进行单细胞培养，获得8个HCCLM3-R-LMI来源的单克隆细胞株（LMI-S2，-S3，-S4，-S5，-S11，-S15，-S21，-S23）和5个HCCLM3-R-LnMI来源的单克隆细胞株（LnM1-S7，-S11，-S13，-S17，-S20）;将上述人肝癌单克隆细胞分别接种于4周龄的裸鼠皮下，3周后皮下瘤组织移植至裸鼠的肝脏，6周后观察裸鼠肺和腹腔淋巴结转移灶荧光面积，并与肺组织连续切片中的转移灶数目进行比较。同一株细胞的肺和淋巴结转移情况比较用Wilcoxon秩和检验，单克隆细胞株之间的肺、淋巴结转移隋况比较用Kruskal-Wallis检验。结果在13株人肝癌单克隆细胞中，有6株细胞形成皮下瘤，移植至裸鼠肝脏后表现出不同的转移潜能和器官靶向特性。其中LM1-S3，LM1-S4，LM1-S5和LM1-S11单克隆细胞的肺转移积分吸光度值分别为80923±10162、1506000±297064、36140±8210和508448±1342729（P〈0．01），但不发生淋巴结转移。LnM1-S11单克隆细胞的肺与淋巴结转移灶积分吸光度值分别为435062±206620和1254000±225171。结论成功建成了不同转移潜能和器官亲嗜性的人肝癌单克隆细胞株和裸鼠移植模型，其中LM1-S3，LM1-S4，LM1-S5和LM1-S11为肺特异亲嗜性的人肝癌单克隆细胞株，而LnM1-S11细胞为肺和淋巴结双重亲嗜性的人肝癌单克隆细胞株，为肝癌转移器官靶向性研究提供了理想的体内外模型。
Objective To establish a single cell-derived organ site-specific metastatic model of human hepatocellular carcinoma （HCC） in the nude mouse. Methods Using the limited dilution method, HCCLM3- R-LM1 and HCCLM3-R-LnM1 cell lines were used to generate eight （LM1-S2, -S3, -S4, -S5, -S11, -S15, -S21, and -S23） and five （LnM 1 -S7, -S 11, -S 13, -S 17, and -S20） single cell-derived monoclonal cell lines, respectively. The monoclonal cell lines were seeded into 4-week-old nude mice, and three weeks later the resultant subcutaneous tumor tissues were orthotopically transplanted into the livers of nude mice. At six weeks after implantation, lung and lymph node were extracted for analysis of the metastatic foci fluorescence area and pathology to assess the number of metastatic foci. Results Among the 13 mice implanted with the established monoclonal cell lines, six grew subcutaneous tumors. When orthotopically transplanted, the six tumors showed remarkably different metastatic potential and organ site-specific tropism. The fluorescence areas of lung metastatic foci were： LM1-S3, 80923 ±10162; LM1-S4, 1506000±297 064;LM1-S5, 36 140±8 210; and LM1-S 11,508 448 ± 134 272 （P〈 0.01）; no lymph node metastases were found for these lines. For LnM1-S11, the fluorescence areas of lung and lymph node metastatic loci were 435062±206620 and 1 254000±225 171, respectively. Conclusion We successfully established several monoclonal cell lines and nude mouse models of HCC with different metastatic potential and organ tropism. Among them, LM1-S3, LM1-S4, LM1-S5, and LM1-S11 have metastasis organotropism to lung. The LnM1-S11 line exhibits dual metastasis organotropism to lung and lymph node. These monoclonal cell lines and nude mouse models may represent useful tools for study of HCC metastasis organotropism.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2012年第7期532-536,共5页
Chinese Journal of Hepatology
基金
国家自然科学基金（81071904）
上海市科委项目（11140902501）
关键词
癌
肝细胞
肿瘤转移
单克隆
器官倾向性
模型
动物
荧光抗体技术
Carcinoma, hepatocellular
Neoplasm metastasis
Monoclone, organ site-specific tropism
Models, animal
Fluorescent antibody technique
