Characterization of stem cell attributes in human osteosarcoma cell lines
Characterization of stem cell attributes in human osteosarcoma cell lines
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DOI：
10.4161/cbt.8.6.7695
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发表时间：
2009-03-15
影响因子：
3.6
通讯作者：
Lin, Chia-Ying
中科院分区：
文献类型：
--
作者：
Wang, Lin;Park, Paul;Lin, Chia-Ying
骨肉瘤是一种侵袭性原发性骨癌，主要影响儿童和青少年。有价值的诊断指标和治疗剂的开发将通过鉴定和表征有助于其攻击行为的基因来增强。新出现的证据表明，肿瘤内的癌症干细胞亚群与异质性恶性肿瘤的发病机制有关。本研究的目的是探讨人骨肉瘤中干细胞样细胞的特征。使用先前开发的肌球形成测定法培养四种人骨肉瘤细胞系。结果表明，所有人骨肉瘤细胞系均具有形成肉瘤球的能力。OS 99 -1和MG 63细胞中的肉瘤球比Hu 09和Saos-2细胞中的肉瘤球多且多见。此外，干细胞标记Oct 3/4和Nanog在骨肉瘤细胞系中使用RT-PCR和免疫学技术进行了检查。我们首次发现，两种Oct 3/4亚型，Oct 3/4 A和Oct 3/4 B，在所有四种细胞系中表达，尽管它们说明了不同的表达模式。Oct 3/4 A在细胞核中表达，而Oct 3/4 B位于所有四种细胞系中发现的细胞亚群的细胞质中。此外，与Saos-2细胞相比，在OS 99 -1、Hu 09和MG 63细胞中观察到升高的Oct 3/4 A表达，而与其他细胞系相比，在Hu 09中检测到显著更高的Oct 3/4 B表达。此外，在所有细胞系的亚群中检测到Nanog，其中OS 99 -1细胞的表达水平显著高于Hu 09、Saos-2和MG 63细胞。因此，我们的数据支持癌症干细胞假说，这可能对骨肉瘤的临床诊断和治疗具有重要意义。
Osteosarcoma is an aggressive primary bone cancer affecting primarily children and young adolescents. Development of valuable diagnostic indicators and therapeutic agents will be enhanced by the identification and characterization of genes that contribute to its aggressive behavior. Emerging evidence suggests a subpopulation of cancer stem cells within tumors that have been implicated in the pathogenesis of heterogeneous malignant tumors. The purpose of our study was to characterize the stem-like cell population in human osteosarcoma. Four human osteosarcoma cell lines were cultured using a previously developed sarcosphere formation assay. The results showed that all human osteosarcoma cell lines possessed an ability to form sarcospheres. More spherical and frequent sarcospheres were observed in OS99-1 and MG63 cells than in Hu09 and Saos-2 cells. Moreover, stem cell markers Oct3/4 and Nanog were examined in osteosarcoma cell lines using RT-PCR and immunological techniques. For the first time, we showed that two Oct3/4 isoforms, Oct3/4 A and Oct3/4 B, were expressed in all four cell lines although they illustrated different expression patterns. Oct3/4 A was expressed in the nuclei while Oct3/4 B was located in the cytoplasm of a subpopulation of cells found within all four cell lines. Furthermore, elevated expression of Oct3/4 A was seen in the OS99-1, Hu09 and MG63 cells compared to Saos-2 cells while significantly higher expression of Oct3/4 B was detected in Hu09 compared with the other cell lines. In addition, Nanog was detected in a subpopulation of all cell lines, where OS99-1 cells expressed significantly higher levels than Hu09, Saos-2 and MG63 cells. Thus, our data support the cancer stem cell hypothesis, which may have important implications for clinical diagnosis and treatment of osteosarcoma.