Deep Learning-Based Imaging Analysis Reveals Radiation-Induced Bystander Effects on Cancer Cell Migration and the...

Regulating tumor invasion and metastasis is pivotal for improving cancer patient prognosis. While cell migration is a key factor in these processes, the non-targeted effects of chemoradiotherapy on cell motility remain poorly understood. In this study, we employed HeLa-FUCCI cells-a cervical cancer-derived HeLa cell line integrated with the Fluorescent Ubiquitination-Based Cell Cycle Indicator (FUCCI) probe, enabling the visualization of cell cycle phases-to investigate the radiation-induced impacts, including non-targeted effects, on cell migration. To create irradiated (In-field) and non-irradiated (out-of-field) regions, half of the culture dish was shielded with a lead block during irradiation. Cells were then exposed to 2 Gy X-rays, with or without cisplatin. Following irradiation, the cells were subjected to time-lapse imaging at 15 min intervals for 24 h, and the acquired data were analyzed using cell segmentation and tracking algorithms, Cellpose 2.0 and TrackMate 7. Without cisplatin, the migration velocity and total distance traveled of Out-of-field cells were significantly reduced compared to controls, suggesting a suppressive bystander signal. In contrast, with cisplatin treatment, these parameters significantly increased in both In-field and Out-of-field cells. This suggests that chemoradiotherapy may inadvertently enhance tumor cell motility outside the target volume, a critical finding with significant implications for therapeutic outcomes.