A method to culture human alveolar rhabdomyosarcoma cell lines as rhabdospheres demonstrates an enrichment in stemness and Notch signaling.

Autor:	Slemmons KK; Departments of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina., Deel MD; Pediatrics, Duke University School of Medicine, Durham, North Carolina., Lin YT; Pediatrics, Duke University School of Medicine, Durham, North Carolina., Oristian KM; Departments of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina.; Pediatrics, Duke University School of Medicine, Durham, North Carolina., Kuprasertkul N; Duke University, Durham, North Carolina., Genadry KC; Pediatrics, Duke University School of Medicine, Durham, North Carolina., Chen PH; Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, North Carolina., Chi JT; Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, North Carolina., Linardic CM; Departments of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, North Carolina corinne.linardic@duke.edu.; Pediatrics, Duke University School of Medicine, Durham, North Carolina.
Jazyk:	angličtina
Zdroj:	Biology open [Biol Open] 2021 Feb 09; Vol. 10 (2). Date of Electronic Publication: 2021 Feb 09.
DOI:	10.1242/bio.050211
Abstrakt:	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. Competing Interests: Competing interestsThe authors declare no competing or financial interests. (© 2021. Published by The Company of Biologists Ltd.)
Databáze:	MEDLINE
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