Departments of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina.
Pediatrics, Duke University School of Medicine, Durham, North Carolina.
Biol Open. 2021 Feb 9;10(2):bio050211. doi: 10.1242/bio.050211.
The development of three-dimensional cell culture techniques has allowed cancer researchers to study the stemness properties of cancer cells in 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 , and and several receptors and transcriptional regulators in the Notch signaling pathway. FP-RMS rhabdospheres also exhibit functional stemness characteristics including multipotency, increased tumorigenicity , and chemoresistance. This method provides a novel practical tool to support research into FP-RMS stemness and chemoresistance signaling mechanisms.
三维细胞培养技术的发展使得癌症研究人员能够在培养中研究癌症细胞的干性特征。然而，迄今为止，尚未报道过培养 PAX3-FOXO1 融合阳性横纹肌肉瘤（FP-RMS）的方法，这阻碍了识别 FP-RMS 干性所涉及的失调信号通路的努力。在这里，我们首先检查了人 RMS 肿瘤和细胞系中典型干细胞标记物的表达。然后，我们描述了一种将 FP-RMS 细胞系培养成横纹球体的方法，并证明这些球体在经典干性因子和 Notch 信号通路成分的表达上更加丰富。具体而言，FP-RMS 横纹球体中表达增加，并且 Notch 信号通路中的几个受体和转录调节剂也增加了表达。FP-RMS 横纹球体还表现出功能性干性特征，包括多能性、增加的致瘤性和化学抗性。这种方法为研究 FP-RMS 干性和化学抗性信号机制提供了一种新的实用工具。
