Mammosphere cells from high-passage MCF7 cell line show variable loss of tumorigenicity and radioresistance

Mammosphere culture of cancer cell lines is an important approach used for enrichment of stem-like cancer cells (SLCs), but over-subcultured cell lines have been experimentally shown to change properties over time. It remains unclear if mammosphere cells (MSs) derived from high-passage cancer cell lines retain the tumorigenicity and radioresistance seen in MSs from primary or low-passage cell lines. In this study, we report that mammospheres derived from MCF-7 sublines after different passage numbers were consistently enriched for CD44+/CD24−/low cells but were not consistently enriched for tumorigenic and radioresistant cells. The tumorigenicity and radioresistance of MSs were associated with their sphere-forming ability, proliferation ability in vitro, and intracellular reactive oxygen species (ROS) levels. The radioresistant MSs showed significant cell cycle arrest in G2/M phase after X-ray irradiation and expressed higher ataxia telangiectasia mutated (ATM) mRNA levels. These results suggest that MSs from high-passage cancer cell lines were not consistently enriched for stem-like cancer cells with higher tumorigenicity and enhanced radioresistance.