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

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Slemmons, KK; Deel, MD; Lin, Y-T; Oristian, KM; Kuprasertkul, N; Genadry, KC; Chen, P-H; Chi, J-TA; Linardic, CM

Published Date

  • February 9, 2021

Published In

Volume / Issue

  • 10 / 2

PubMed ID

  • 33372065

Pubmed Central ID

  • PMC7888706

Electronic International Standard Serial Number (EISSN)

  • 2046-6390

Digital Object Identifier (DOI)

  • 10.1242/bio.050211


  • eng

Conference Location

  • England