体内连续筛选法建立人结肠癌肝转移阶梯细胞模型  

The establishment of stepwise metastatic colorectal carcinoma cell model to the liver via in vivo selection

作　　者：万方军[1] 蔡健华[1] 赵任[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院普外科,上海200025

出　　处：《外科理论与实践》2009年第6期633-638,共6页Journal of Surgery Concepts & Practice

基　　金：国家自然科学基金课题(30772106)

摘　　要：目的:通过在裸鼠体内进行SW1116结肠癌细胞株原位种植肝转移连续筛选,建立人结肠癌肝转移阶梯细胞模型系统,为研究结肠癌转移的分子机制提供实验模型。方法:将SW1116细胞株在裸鼠皮下连续5次传代后,取皮下种植瘤在裸鼠结肠进行原位种植,继而取肝脏转移灶进行下一代裸鼠结肠原位种植,经3次结肠原位种植肝转移筛选,建立大肠癌肝转移阶梯细胞模型。用免疫组化法验证种植瘤及转移灶的组织来源,并行同期体内外侵袭实验,检测经体内连续筛选出的癌细胞侵袭转移能力变化。结果:经5代皮下连续传代及3代结肠至肝的体内转移筛选,成功建立了第三代结肠癌肝转移筛选细胞系(CRCLM3)。裸鼠种植瘤及转移灶均表达人癌胚抗原,而同期体内外侵袭试验结果显示,经体内筛选得到的结肠癌细胞株转移能力逐渐加强。结论:经体内连续肝转移筛选所建立的结肠癌细胞系侵袭转移能力增强,而CRCLM3细胞系是研究结肠癌肝转移机制的理想细胞模型。Objective To establish a stepwise metastatic cell line model of colorectal carcinoma to the liver via a consecutive in vivo selection with the SW1116 cell line, which would be used on the study of the molecular mechanism of metastasis of eolorectal carcinoma. Methods SW1116 cells were transplanted into the subcutis of nude mice and serially passaged for 5 times, then the subcutaneous tumor was implanted into the cecal wall of new nude mice. The liver metastastie lesions were picked out and implanted into the cecal wall of new nude mice; by implanting 3 times, stepwise metastatic colorectal carcinoma cell model was established by in vivo selection; The expression of human carcinoma antigen CEA in the subcutaneous tumor tissues, cecal tumor tissues and liver metastatic tissues were investigated by immunohistochemistry to prove that they were of human origin. In vivo and in vitro tests were performed to know the difference of its invasiveness and metastatic potential after in vivo selection. Results After 5 times of subcutaneous continuous passage and 3 screening process of consecutive hepatic metastases in vivo selection, the colorectal carcinoma liver metastasis selection 3（CRCLM3） cell line was established. Human carcinoembryonic antigen was detected in tumors from the nude mice. In vivo and in vitro tumor invasion and migration assay demonstrated that there was significant difference in the ability of invassiveness and metastatic potential in cell lines after in vivo selection. Conclusions CRCLM3 cell line could be established via consecutive in vivo selection with the SWlll6 cell line. The new cell line has stronger invasiveness and higher metastatic potential than the original SW1116 cell. The CRCLM3 cell model could be used on the study of the molecular mechanism of metastasis of colorectal carcinoma.

关 键 词：大肠癌 原位种植 阶梯转移模型 SW1116细胞株 实验模型 

分 类 号：R735.35[医药卫生—肿瘤]