CAJ | 학술논문

目的：观察肿瘤干细胞标志物P75NTR、Oct-4、Sox-2、Lin28、Nanog在食管鳞癌Eca109细胞系富集的细胞球细胞中的表达。探讨食管鳞癌的肿瘤干细胞标记物。方法采用无血清成球培养法，富集培养细胞球细胞，观察其增殖过程。培养5d获得小细胞球继续培养至14d获得又大又圆的悬浮生长的细胞球，收集第14天的细胞球，一部分用于实验，一部分予以传代。应用免疫荧光技术检测细胞球细胞中P75NTR、Oct-4、Sox-2、Lin28、Nanog的表达和定位。结果 P75NTR、Oct-4荧光定位于细胞球细胞的细胞核上，贴壁的Eca109为细胞核和细胞质表达，但是Oct-4荧光强度比P75NTR弱；Lin28在细胞球细胞上为细胞核表达，贴壁的Eca109上为细胞核和细胞质表达；Nanog和Sox-2在食管鳞癌Eca109细胞球细胞和贴壁的Eca109上均为细胞质表达。结论 P75NTR、Oct-4、Lin28核表达阳性的细胞球可能为食管癌干细胞。
목적：관찰종류간세포표지물P75NTR、Oct-4、Sox-2、Lin28、Nanog재식관린암Eca109세포계부집적세포구세포중적표체。탐토식관린암적종류간세포표기물。방법채용무혈청성구배양법，부집배양세포구세포，관찰기증식과정。배양5d획득소세포구계속배양지14d획득우대우원적현부생장적세포구，수집제14천적세포구，일부분용우실험，일부분여이전대。응용면역형광기술검측세포구세포중P75NTR、Oct-4、Sox-2、Lin28、Nanog적표체화정위。결과 P75NTR、Oct-4형광정위우세포구세포적세포핵상，첩벽적Eca109위세포핵화세포질표체，단시Oct-4형광강도비P75NTR약；Lin28재세포구세포상위세포핵표체，첩벽적Eca109상위세포핵화세포질표체；Nanog화Sox-2재식관린암Eca109세포구세포화첩벽적Eca109상균위세포질표체。결론 P75NTR、Oct-4、Lin28핵표체양성적세포구가능위식관암간세포。
Objective To observe the expression of tumor stem cell markers P 75NTR,Oct-4,Sox-2,Lin28 and Nanog in the tumor sphere from esophageal squamous cells carcinoma Eca 109 and identify the esophageal squamous cell cancer stem cell marker .Methods The serum-free culture method was used for generating tumor spheres: proliferation was observed in enrichment culture tumor spheres .Small tumor spheres were obtained after 5 days culture and big and round tumor spheres appeared after 14 days culture which were collected for experiments and passaged .The expression and location of P75NTR,Oct-4,Sox-2,Lin28, and Nanog were detected by immunofluorescence cytochemistry .Results The expressions of P75NTR,Oct-4 and Lin28 were positive in the center of tumor spheres and some on cytoplasm and other in nuclei of Eca109 monolayer cells.However, Oct-4 fluorescence intensity was weaker than P75NTR.The expressions of Sox-2 and Nanog were positive in cytoplasm of tumor spheres and Eca 109 monolayer cells .Conclusion The cells expressing P75NTR, Oct-4, and Lin28 in the center of the tumor sphere may be esophageal cancer stem cells .