人食管癌细胞株KYSE-150、TE-1中肿瘤干细胞的培养分化及增殖侵袭能力分析

Culture, differentiation, proliferation and invasion of tumor stem cells in human esophageal carcinoma cell lines KYSE-150 and TE-1

背景:研究发现,食管癌组织中存在一类细胞亚群,具有一定的侵袭和转移性能,与治疗效果之间存在十分密切的联系。目的:从人食管癌细胞株KYSE-150、TE-1中分离富含肿瘤干细胞的细胞球,并对其增殖和侵袭能力进行分析。方法:运用无血清培养基培养KYSE-150、TE-1细胞,观察细胞球的生成情况,采用MTT法以及Transwell小室实验检测细胞的增殖情况和侵袭能力,流式细胞仪检测细胞表面标志物的表达。结果与结论:①KYSE-150、TE-1细胞在无血清培养条件下可以获得稳定传代的细胞球。②细胞球的增殖能力和侵袭能力均显著高于亲本细胞(P<0.05)。③TE-1细胞球和KYSE-150细胞球的CD44^+、CD271^+、CD44^+CD271^+细胞比例均显著高于TE-1亲本细胞和KYSE-150亲本细胞(P<0.05)。④实验结果提示,从人食管癌细胞株KYSE-150、TE-1中分离出具有肿瘤干细胞特性的细胞球,具有很强的增殖和侵袭转移能力,且CD44和CD271可以作为重要的食管癌干细胞表面标志物。

BACKGROUND： There is a certain cell subset in esophageal cancer tissues, with certain invasive and metastatic properties, which is closely related to the clinical therapeutic effect on tumors.OBJECTIVE： To isolate tumor stem cell spheres in human esophageal carcinoma cell lines KYSE-150 and TE-1 and to analyze their proliferation and invasion ability. METHODS： KYSE-150 and TE-1 cells were cultured in serum-free medium to observe the formation of cell spheres. Cell proliferation and invasion were detected using MTT and Transwell chamber culture. Surface markers of cells were detected using flow cytometry. RESULTS AND CONCLUSION： Cell spheres that were stably subcultured were obtained from KYSE-150 and TE-1 cells cultured in serum-free medium. The proliferation and invasion abilities of cell spheres were significantly stronger than those of parent cells（P〈0.05）. The number of CD44＋, CD271＋ and CD44＋CD271＋ cells in TE-1 and KYSE-150 cell spheres was significantly higher than that in the TE-1 and KYSE-150 parent cells（P〈0.05）. These experimental results show that cell spheres isolated from human esophageal carcinoma cell lines TE-1 and KYSE-150 have tumor stem cell properties as well as strong proliferation and invasion abilities. And moreover, CD44 and CD271 can be used as important surface markers of esophageal carcinoma stem cells.