Ferroptosis-dependent breast cancer cell-derived exosomes inhibit migration and invasion of breast cancer cells by suppressing M2 macrophage polarization.

ABSTRACT

Aim: Ferroptosis, a novel type of iron-dependent cell death, plays a vital role in breast cancer progression. However, the function of ferroptosis-induced cancer cell-derived exosomes in breast cancer remains unclear. In this study, we attempted to investigate the impact of breast cancer cells-derived exosomes induced by ferroptosis on the polarization of macrophages and the progression of breast cancer. Methods: Erastin was used to induce ferroptosis and breast cancer cell-derived exosomes were identified by transmission electron microscopy. Western blot, quantitative reverse transcription PCR, immunofluorescence, flow cytometry, and ELISA were used to determine the role of exosomes in macrophage polarization. Transwell assays were used to detect breast cancer cell migration, and invasion. Results: Our results showed that erastin promoted ferroptosis in breast cancer cells with increased Fe2+ level and ROS production. Breast cancer cell-derived exosomes induced by ferroptosis were successfully isolated and verified to be internalized by macrophages. In addition, ferroptosis-induced breast cancer cell-derived exosomes (Fe-exo) remarkably diminished M2 marker, Arg-1 expression. The ratio of CD206+ macrophages was significantly decreased after Fe-exo treatment. CD206 protein expression and Arg-1 level were dramatically reduced in M2 macrophages incubated by Fe-exo. Moreover, autophagy PCR array showed that the expression of 84 autophagy-related genes were altered after macrophages were incubated by Fe-exo. Furthermore, macrophages incubated by Fe-exo repressed the migration and invasion of breast cancer cells. Conclusion: Ferroptosis-dependent cancer cell-derived exosomes inhibited M2 polarization of macrophages, which in turn inhibited migration and invasion of breast cancer cells. This study provides novel therapeutic strategies for patients with breast cancer.