高转移潜力食管癌细胞株的建立及其生物学特征

Establishment and Characterization of a Cell Line with High Metastatic Potential Derived from Human Esophageal Carcinoma Cell Line EC199

目的建立具有高转移潜力食管癌细胞株并研究其生物学特征。方法将食管癌细胞系EC109细胞悬液异位移植到SCID小鼠胃壁,约3个月后或动物濒临死亡时处死,行病理学解剖,将肉眼可见的纵隔淋巴结转移瘤块接种于SCID鼠皮下扩增,然后取小鼠皮下瘤组织块进行细胞培养,得到性状稳定的细胞株NMC109后,用MTT法分析细胞生长曲线,Western bloting法检测与细胞分裂增殖能力密切相关的TopoIIα表达,酶谱法检测MMP-2和MMP-9的活性,划痕实验和Transwell体外移动实验检测细胞的移动能力。结果与母本细胞EC109相比,所获得的细胞株NMC109其增殖能力和TopoIIα表达明显增强,MMP-9的活性明显升高,移动能力明显增强。结论获得了具有高转移潜力的食管癌细胞株。

Objective To evaluate the characteristics of a cell line derived from human esophageal carcinoma cell line EC109. Method EC109 cells were heterotopically inoculated into the gastric wall of SCID mice, and the mice were sacrificed after 3 months or when a mouse was dying. The mice were examined and the metastatic mediastinal lymph nodes were taken and transplanted subcutaneously in SCID mice. After that, tumor tissue was taken from those subcutaneous tumors and primary cell culture was performed to establish a stable cell strain. Cell proliferation was determined by M＇IT assay, and the expression of TopolIu was determined by Western blotting. The activity of MMP-9 and MMP-2 was measured by zymography. Finally,migration ability of the cells was evaluated by Transwell method and wound-healing method. Results A highly metastatic cell line was established and named NMC109, and demonstrated that the proliferation, migration and MMP-9 activity of NMC109 cells were increased. Conclusion The cell line NMC109 can be employed to obtain more aggressive cells with metastatic potential, providing an useful basis for further research works.