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多种肿瘤干细胞标志物在食管鳞癌Eca109细胞球细胞中的表达
黄燕燕 裴学莲 刘洋 李秋霞 王玉婷 慕晓玲*
解剖学报 ›› 2014, Vol. 45 ›› Issue (3) : 350-353.
多种肿瘤干细胞标志物在食管鳞癌Eca109细胞球细胞中的表达
Expression of tumor stem cell markers in the tumor sphere from esophageal squamous cells carcinoma Eca109
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。
Objectiv To observe the expression of tumor stem cell markers P75NTR,Oct-4,Sox-2,Lin28 and Nanog in the tumor sphere from esophageal squamous cells carcinoma Eca109 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 Eca109 monolayer cells. Conclusion The cells expressing P75NTR, Oct-4, and Lin28 in the center of the tumor sphere may be esophageal cancer stem cells.
P75NTR / Oct-4 / Sox-2；Lin28 / Nanog / 肿瘤干细胞 / 食管鳞癌Eca109细胞 / 无血清成球培养法
P75NTR / Oct-4 / Sox-2 / Lin28 / Nanog / Cancer stem cell / Esophageal squamous cell carcinoma / Tumor sphere formation
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