Human Rhabdomyosarcoma Cell Lines for Rhabdomyosarcoma Research: Utility and Pitfalls

Abstract

Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma of childhood and adolescence. Despite intergroup clinical trials conducted in Europe and North America, outcomes for high risk patients with this disease have not significantly improved in the last several decades, and survival of metastatic or relapsed disease remains extremely poor. Accrual into new clinical trials is slow and difficult, so in vitro cell-line research and in vivo xenograft models present an attractive alternative for preclinical research for this cancer type. Currently, 30 commonly used human RMS cell lines exist, with differing origins, karyotypes, histologies, and methods of validation. Selecting an appropriate cell line for RMS research has important implications for outcomes. There are also potential pitfalls in using certain cell lines including contamination with murine stromal cells, cross-contamination between cell lines, discordance between the cell line and its associated original tumor, imposter cell lines, and nomenclature errors that result in the circulation of two or more presumed unique cell lines that are actually from the same origin. These pitfalls can be avoided by testing for species-specific isoenzymes, microarray analysis, assays for subtype-specific fusion products, and short tandem repeat analysis.

Helicase CHD4 is an epigenetic coregulator of PAX3-FOXO1 in alveolar rhabdomyosarcoma

“…All cells were cultured in 5% CO 2 at 37°C. All RMS and the MRC5 cell line were authenticated by short tandem repeat analysis (STR) profiling in 2014/2015 and positively matched (71). As the human myoblast cell line has not yet been characterized by STR profiling, negative matching with all available cell lines in the database was used for verification.…”

Uncovering metabolism in rhabdomyosarcoma

“…74 In response to hypoxia, an increased glucose uptake was also observed in the A204 cell line to be protective against cell death 77 ; however, it should be noted that this cell line is no longer retained as truly representative of RMS. 78 Finallly, the cardiac-specific deletion of the von Hippel-Lindau protein (VHL), an E3 ubiquitin ligase responsible of suppression of HIF levels during normoxia, has been described to favor the formation of highly metastatic cardiac RMS in a mouse model. 79 Taken together, different lines of evidence indicate that the PI3K/AKT and ERK1/2 pathways, often concurrently with the HIF pathway, play a central role for RMS progression; their targeted inhibition was indeed shown to enhance cell death in RMS lines under both normoxic 80 and hypoxic conditions.…”

“…42,89 P53 gene inactivation is indeed an experimentally validated cooperating lesion in zebrafish and mouse models of ERMS 29 and ARMS, 25 respectively. In the human RD cell line, one of the most widely used human ERMS cell lines, 78 loss of p53 activity was reported to rise the glucose uptake by increased expression of the GLUT1 and GLUT4 transporters 83 (Fig. 3).…”

PLK1 Phosphorylates PAX3-FOXO1, the Inhibition of Which Triggers Regression of Alveolar Rhabdomyosarcoma

“…aRMS and eRMS cell lines were tested and authenticated by cell line typing analysis (STR profiling) in 2011/2014 and positively matched (32).…”