Notch2表达和胰腺癌干细胞相关.docVIP

肿瘤 2011 年 8 月第 31 卷第 8 期　Tumor Vol.31, August 2011　　 基础研究·Basic Research  707 DOI: 10.3781/j.issn.1000-7431.2011.08.006 Copyright? 2011 by TUMOR  Notch2表达与胰腺癌干细胞相关 周竹超1，董强刚2，倪泉兴3 1. 复旦大学附属华山医院普外科，上海　200040 ；2. 上海市肿瘤研究所肿瘤干细胞研究组，上海　200032 ；3. 复 旦大学附属肿瘤医院胰腺外科，上海　200032 [ 摘要 ]　目的：以 Notch2 为表面标志对胰腺癌干细胞进行分离鉴定。方法：采用磁性细胞分选（magnetic activated cell sorting，MACS）技术纯化胰腺癌 Bxpc-3 和 Panc-1 细胞株中的 Notch2 ＋细胞，通过 NOD-SCID 小鼠皮下移植 评估其致瘤能力。在无血清条件下培养观察细胞的球体形成能力，应用免疫荧光检测法检测干细胞相关标志八聚体 结合转录因子 4（octamer-binding transcription factor 4，Oct4）、Nanog 和胰十二指肠同源异型盒 1（pancreatic and duodenal homeobox 1，PDX1）的表达，并通过测序检测其 K-ras 基因突变的情况。结果：Notch2 ＋细胞具有显著致 瘤性。胚胎干细胞核心转录因子 Oct4、Nanog 和胰腺干细胞标志 PDX1 阳性表达。在无血清条件下培养能形成细胞 球体，并存在 K-ras 基因的突变。结论：人胰腺癌 Notch2 ＋ Bxpc-3 和 Panc-1 细胞具有肿瘤干细胞特性。 [ 关键词 ]　胰腺肿瘤；肿瘤干细胞；基因表达；细胞，Bxpc-3，Panc-1 [文献标志码] Notch2 expression in association with pancreatic cancer stem cells ZHOU Zhu-chao1, DONG Qiang-gang2, NI Quan-xing3 1. Department of General Surgery, Huashan Hospital, Fudan University, Shanghai 200040, China; 2. Laboratory of Cancer Stem Cells, Shanghai Cancer Institute, Shanghai 200032, China; 3. Department of Pancreatic Surgery, Cancer Hospital, Fudan University, Shanghai 200032, China [ABSTRACT] Objective: To isolate and identify the pancreatic cancer stem cells based on surface marker Notch2. Methods: The Notch2＋ cells in pancreatic cancer Bxpc-3 and Panc-1 cells were separated by magnetic activated cell sorting (MACS) technique. The tumorigenic potent of these cells was evaluated via transplantation of the cells into NOD-SCID mice. Notch2＋ cells were cultured with serum-free medium to detect the tumorsphere formation ability. The expressions of stem cell-related markers—octamer-binding transcription factor 4 (Oct4), Nanog and pancreatic stem cell marker pancreatic and duodenal homeobox 1 (PDX1) in the isolated cells were measured by immunofluorescent staining. T