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
Language
- eng
Conference Location
- England