Side Population is Not Necessary or Sufficient for a Cancer Stem Cell Phenotype in Glioblastoma Multiforme
Side Population is Not Necessary or Sufficient for a Cancer Stem Cell Phenotype in Glioblastoma Multiforme
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DOI：
10.1002/stem.582
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发表时间：
2011-03-01
期刊：
影响因子：
5.2
通讯作者：
McConnell, Melanie J.
中科院分区：
文献类型：
--
作者：
Broadley, Kate W. R.;Hunn, Martin K.;McConnell, Melanie J.
有强有力的证据表明，在侵袭性脑肿瘤多形性胶质母细胞瘤（GBM）中存在癌症干细胞（CSC）。这些细胞具有干细胞样的自我更新活性和增加的肿瘤起始能力，并且被认为是由于它们对治疗的抗性而导致复发的原因。已经使用了几种技术来富集CSC，包括在无血清限定培养基中生长以诱导球体形成，以及使用排除荧光染料Hoechst 33342（侧群（SP））分离干细胞样细胞。我们表明，球形成GBM细胞系和原代GBM细胞富集CSC样表型的自我更新基因表达增加体外和增加体内肿瘤的起始。然而，SP在所有球形培养物中均不存在。从GBM系中直接分离SP在体外没有富集干细胞样活性，并且与非SP和亲本细胞相比，分选的SP的肿瘤起始活性较低。短暂暴露于阿霉素增强CSC和SP频率。然而，阿霉素治疗改变了细胞计数的轮廓和模糊的SP证明在压力下的细胞中识别SP的困难。多柔比星暴露的细胞表现出SP的短暂增加，但多柔比星-SP细胞仍然没有富集干细胞样自我更新表型。这些数据表明，GBM SP不一定有助于自我更新或肿瘤起始，CSC的关键特性，我们建议不要使用SP计数或分离CSC。干细胞2011;29：452-461
There is strong evidence for the existence of cancer stem cells (CSCs) in the aggressive brain tumor glioblastoma multiforme (GBM). These cells have stem-like self-renewal activity and increased tumor initiation capacity and are believed to be responsible for recurrence due to their resistance to therapy. Several techniques have been used to enrich for CSC, including growth in serum-free defined media to induce sphere formation, and isolation of a stem-like cell using exclusion of the fluorescent dye Hoechst 33342, the side population (SP). We show that sphere formation in GBM cell lines and primary GBM cells enriches for a CSC-like phenotype of increased self-renewal gene expression in vitro and increased tumor initiation in vivo. However, the SP was absent from all sphere cultures. Direct isolation of the SP from the GBM lines did not enrich for stem-like activity in vitro, and tumor-initiating activity was lower in sorted SP compared with non-SP and parental cells. Transient exposure to doxorubicin enhanced both CSC and SP frequency. However, doxorubicin treatment altered the cytometric profile and obscured the SP demonstrating the difficulty of identifying SP in cells under stress. Doxorubicin-exposed cells showed a transient increase in SP, but the doxorubicin-SP cells were still not enriched for a stem-like self-renewal phenotype. These data demonstrate that the GBM SP does not necessarily contribute to self-renewal or tumor initiation, key properties of a CSC, and we advise against using SP to enumerate or isolate CSC. STEM CELLS 2011;29:452-461