How Stemlike Are Sphere Cultures From Long-term Cancer Cell Lines? Lessons From Mouse Glioma Models
How Stemlike Are Sphere Cultures From Long-term Cancer Cell Lines? Lessons From Mouse Glioma Models
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DOI：
10.1097/nen.0000000000000130
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发表时间：
2014-11-01
影响因子：
3.2
通讯作者：
Weller，Michael
中科院分区：
文献类型：
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作者：
Ahmad，Mushfika;Frei，Karl;Weller，Michael
癌症干细胞可以介导各种类型癌症（包括胶质母细胞瘤）的治疗抗性和复发。癌症干细胞样细胞可以从长期癌细胞系（包括神经胶质瘤系）中分离。使用球体形成作为体外癌细胞干细胞性的模型，我们从SMA-497、SMA-540、SMA-560和GL-261胶质瘤细胞中获得球体培养物。基因表达和蛋白质组学分析表明，球形培养物一致地显示出干细胞相关基因的表达升高，特别是包括CD 44。非球形和球形培养物之间神经谱系标志物表达的差异是异质性的，除了球形培养物中β-III-微管蛋白的均匀减少。所有球形培养物均显示生长较慢。自我更新能力的影响，培养基条件，但不nonsphere与球文化表型。球形培养物对辐射更具抵抗力，而非球形和球形培养物对替莫唑胺具有高度抵抗力。在除SMA-560外的所有模型中，非球形细胞在同基因小鼠中形成的肿瘤比球形细胞更具侵袭性。植入同基因宿主的非球形和球形细胞来源的肿瘤之间，T细胞或小胶质细胞/巨噬细胞的血管形成或浸润没有重大差异。总之，这些数据表明，小鼠神经胶质瘤细胞系可以在体外诱导形成获得干性特征的球体，但它们不表现出均匀的生物表型，从而挑战了它们代表上级模型系统的观点。
Cancer stem cells may mediate therapy resistance and recurrence in various types of cancer, including glioblastoma. Cancer stemlike cells can be isolated from long-term cancer cell lines, including glioma lines. Using sphere formation as a model for cancer cell stemness in vitro, we derived sphere cultures from SMA-497, SMA-540, SMA-560, and GL-261 glioma cells. Gene expression and proteomics profiling demonstrated that sphere cultures uniformly showed an elevated expression of stemness-associated genes, notably includingCD44. Differences in neural lineage marker expression between nonsphere and sphere cultures were heterogeneous except for a uniform reduction of β-III-tubulin in sphere cultures. All sphere cultures showed slower growth. Self-renewal capacity was influenced by medium conditions but not nonsphere versus sphere culture phenotype. Sphere cultures were more resistant to irradiation, whereas both nonsphere and sphere cultures were highly resistant to temozolomide. Nonsphere cells formed more aggressive tumors in syngeneic mice than sphere cells in all models except SMA-560. There were no major differences in vascularization or infiltration by T cells or microglia/macrophages between nonsphere and sphere cell–derived tumors implanted in syngeneic hosts. Together, these data indicate that mouse glioma cell lines may be induced in vitro to form spheres that acquire features of stemness, but they do not exhibit a uniform biologic phenotype, thereby challenging the view that they represent a superior model system.