CAJ | 학술논문

为分离富集人食管鳞癌Eca109细胞株中肿瘤干细胞，并对其“干细胞”特性进行鉴定。采用无血清（DMEM/F121∶1，无血清）成球培养法获得细胞球，应用免疫荧光细胞化学技术检测Eca109细胞株中细胞球细胞中干性相关转录因子的表达；平板克隆和MTT法检测细胞体外增殖情况；Transwell小室法检测其细胞侵袭能力。结果显示，无血清培养4 d获得Eca109细胞球；P75NTR和Oct-4荧光定位于细胞球细胞的细胞核上，Eca109细胞为细胞核和细胞浆表达，但是Oct-4荧光强度比P75NTR弱；MTT法测增殖显示细胞细胞球增殖率明显高于Eca109细胞，差异有统计学意义（P＜0.001）；平板克隆实验显示细胞球细胞形成克隆团数高于Eca109细胞；Transwell小室法结果显示细胞球细胞侵袭力高于Eca109细胞。由此可知，食管鳞癌Eca109细胞系通过无血清成球培养法获得细胞球，细胞球存在肿瘤干细胞。
위분리부집인식관린암Eca109세포주중종류간세포，병대기“간세포”특성진행감정。채용무혈청（DMEM/F121∶1，무혈청）성구배양법획득세포구，응용면역형광세포화학기술검측Eca109세포주중세포구세포중간성상관전록인자적표체；평판극륭화MTT법검측세포체외증식정황；Transwell소실법검측기세포침습능력。결과현시，무혈청배양4 d획득Eca109세포구；P75NTR화Oct-4형광정위우세포구세포적세포핵상，Eca109세포위세포핵화세포장표체，단시Oct-4형광강도비P75NTR약；MTT법측증식현시세포세포구증식솔명현고우Eca109세포，차이유통계학의의（P＜0.001）；평판극륭실험현시세포구세포형성극륭단수고우Eca109세포；Transwell소실법결과현시세포구세포침습력고우Eca109세포。유차가지，식관린암Eca109세포계통과무혈청성구배양법획득세포구，세포구존재종류간세포。
Objective to isolate and characterize the cancer stem cells from esophageal sequamous cell carcinoma cell line Eca109 using mammosphere formation.Methods Serum -free culture method was used to form a mammosphere;Immunofluorescence cytochemistry was used to detect the expression of stemness relative transcriptional factors of the spheres;The cell proliferation level was tested by MTT and tablet cloning;Invasive ability of each cell line was detected by transwell chamber assay.Results Mammospheres were generated at day 4.P75NTR and Oct-4 were located in nuclear of Eca109 mammosphere cells.However, Oct-4 fluorescence intensity was weaker than that of P75NTR;it was showed by MTT assay that the mammosphere proliferation rate was obviously higher than Eca109 cells,and the difference was statistically significant (P<0.001).Tablet cloning experiment showed that mammospheres cell colony forming was higher than that of Eca109 cells,Transwell assay showed the invasion of Eca109 mammospheres was higher than that of Eca109 cells.Conclusion Eca109 cells generate tumor spheres under serum-free stem cell medium,in which cancer stem cells were existed.