A method to culture human alveolar rhabdomyosarcoma cell lines as rhabdospheres demonstrates an enrichment in stemness and Notch signaling.
A method to culture human alveolar rhabdomyosarcoma cell lines as rhabdospheres demonstrates an enrichment in stemness and Notch signaling.
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DOI：
10.1242/bio.050211
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发表时间：
2021-02-09
期刊：
影响因子：
2.4
通讯作者：
Linardic CM
中科院分区：
文献类型：
--
作者：
Slemmons KK;Deel MD;Lin YT;Oristian KM;Kuprasertkul N;Genadry KC;Chen PH;Chi JT;Linardic CM
三维细胞培养技术的发展使癌症研究人员能够在体外培养中研究癌细胞的干性特性。然而，一种培养PAX3-FOXO1融合阳性横纹肌肉瘤（FP-RMS）的方法，一种侵袭性的儿童软组织肉瘤，迄今尚未报道，阻碍了识别FP-RMS发病基础的失调信号通路的努力。在这里，我们首先研究了典型干细胞标记物在人类RMS肿瘤和细胞系中的表达。然后，我们描述了一种将FP-RMS细胞系培养成横纹肌球的方法，并证明这些球富含典型干性因子和Notch信号成分的表达。具体来说，FP-RMS横纹肌球增加了SOX2、POU5F1 （OCT4）和NANOG以及Notch信号通路中几种受体和转录调节因子的表达。FP-RMS横纹肌球也表现出功能性干性特征，包括多能性、体内致瘤性增强和化疗耐药。该方法为研究FP-RMS的干性和耐药信号机制提供了一种新的实用工具。摘要：本文报道了一种三维培养人PAX3-FOXO1融合阳性横纹肌肉瘤细胞的方法，并将这些横纹肌球作为研究其干细胞性和耐药信号机制的新工具。
The development of three-dimensional cell culture techniques has allowed cancer researchers to study the stemness properties of cancer cells in in vitro culture. However, a method to grow PAX3-FOXO1 fusion-positive rhabdomyosarcoma (FP-RMS), an aggressive soft tissue sarcoma of childhood, has to date not been reported, hampering efforts to identify the dysregulated signaling pathways that underlie FP-RMS stemness. Here, we first examine the expression of canonical stem cell markers in human RMS tumors and cell lines. We then describe a method to grow FP-RMS cell lines as rhabdospheres and demonstrate that these spheres are enriched in expression of canonical stemness factors as well as Notch signaling components. Specifically, FP-RMS rhabdospheres have increased expression of SOX2, POU5F1 (OCT4), and NANOG, and several receptors and transcriptional regulators in the Notch signaling pathway. FP-RMS rhabdospheres also exhibit functional stemness characteristics including multipotency, increased tumorigenicity in vivo, and chemoresistance. This method provides a novel practical tool to support research into FP-RMS stemness and chemoresistance signaling mechanisms. Summary: Here we report on a method to culture human PAX3-FOXO1 fusion-positive rhabdomyosarcoma cells in three dimensions, and use these rhabdospheres as a novel tool to study their stemness and chemoresistance signaling mechanisms.