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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 factor4,Oct4)、Nanog和胰十二指肠同源异型盒1(pancreatic and duodenal homeobox1,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。
Objective: To isolate and identify pancreatic cancer stem cells with Notch2 as the surface marker. Methods: Notch2 + cells from pancreatic cancer cell lines Bxpc-3 and Panc-1 were purified by magnetic activated cell sorting (MACS). The tumorigenicity was evaluated by subcutaneous transplantation of NOD-SCID mice. The spheroid formation ability of the cells was observed under the condition of serum-free culture. The stem cell-associated markers octamer-binding transcription factor 4 (Oct4), Nanog and pancreaticoduodenal homeobox 1 in pancreatic and duodenal homeobox1 (PDX1), and detect the mutation of K-ras gene by sequencing. Results: Notch2 + cells have significant tumorigenicity. Embryonic stem cell core transcription factor Oct4, Nanog and pancreatic stem cell marker PDX1 positive expression. Culturing in serum-free conditions formed cell spheres with mutations in the K-ras gene. Conclusions: Notch2 + Bxpc-3 and Panc-1 cells of human pancreatic cancer have tumor stem cell characteristics.