Subclinical dose irradiation triggers human breast cancer migration via mitochondrial reactive oxygen species

Background: Despite technological advances in radiotherapy, Cancer cells at the tumor margin and in diffusive infiltrates can receive subcytotoxic doses of photons. Even if only a minority of Cancer cells are concerned, phenotypic consequences could be important considering that mitochondrial DNA (mtDNA) is a primary target of radiation and that damage to mtDNA can persist. In turn, mitochondrial dysfunction associated with enhanced mitochondrial ROS (mtROS) production could promote Cancer cell migration out of the irradiation field in a natural attempt to escape therapy. In this study, using MCF7 and MDA-MB-231 human breast Cancer cells as models, we aimed to elucidate the molecular mechanisms supporting a mitochondrial contribution to Cancer cell migration induced by subclinical doses of irradiation (< 2 Gy).

Methods: Mitochondrial dysfunction was tested using mtDNA multiplex PCR, oximetry, and ROS-sensitive fluorescent reporters. Migration was tested in transwells 48 h after irradiation in the presence or absence of inhibitors targeting specific ROS or downstream effectors. Among tested inhibitors, we designed a mitochondria-targeted version of human catalase (mtCAT) to selectively inactivate mitochondrial H₂O₂.

Results: Photon irradiation at subclinical doses (0.5 Gy for MCF7 and 0.125 Gy for MDA-MB-231 cells) sequentially affected mtDNA levels and/or integrity, increased mtROS production, increased MAP2K1/MEK1 gene expression, activated ROS-sensitive transcription factors NF-κB and AP1 and stimulated breast Cancer cell migration. Targeting mtROS pharmacologically by MitoQ or genetically by mtCAT expression mitigated migration induced by a subclinical dose of irradiation.

Conclusion: Subclinical doses of photon irradiation promote human breast Cancer migration, which can be countered by selectively targeting mtROS.

Breast cancer; Catalase; Migration; MitoQ; Mitochondria; Reactive oxygen species (ROS); Subclinical dose irradiation.