The Effects Of Macrophage Polarization By Ferroptosis Inducer Erastin In Ovarian Cancer Cells

OBJECTIVE: To investigate the effect and mechanism of ferroptosis inducer erastin on ovarian cancer cells by regulating the polarization of macrophages.Method: 1.Human mononuclear cell line THP-1 induced by PMA differentiated into M0 macrophages;M0 macrophages were treated with erastin at different concentrations for 24 h,and the sensitivity of macrophages to erastin-induced ferroptosis was detected by cck-8;2.10μM erastin stimulate M0 macrophages for 24 hours,then detect the expression of M1 and M2 macrophage markers by q RT-PCR,flow cytometry,and immunofluorescence;3.Human peripheral blood mononuclear cells(peripheral blood mononuclear cells,PBMC)-derived monocytes were induced by MCSF for 6 days,and the adherent cells were M0 macrophages.24 hours after erastin treatment of M0 macrophages,q RT-PCR and flow cytometry were used to detect the expression of M2 macrophage markers;4.Western Blot detected the protein levels of PSTAT3,STAT3,P-STAT6,and STAT6 in macrophages after erastin stimulation;5.STAT3 pathway inhibitor Statistic was used to treat macrophages,and Western Blot was used to confirm its inhibition of the phosphorylation of STAT3;q RT-PCR and flow cytometry was performed to detect the expression of M2 markers in macrophages;6.The conditioned medium of erastin-stimulated macrophages was collected,migration and invasion ability of EOC cells SKOV3 and HO8910 cultivated with different conditioned medium was detected by wound healing and transwell assay.Results:1.Erastin at a concentration of 5-10 μM would not cause significant cell death of THP-1 derived macrophages,so 10μM was selected as experimental concentration.2.After treatment with erastin,there was no significant change in the expression level of M1 macrophage markers,and the expression level of M2 macrophage markers increased significantly in THP-1 derived macrophages.3.Erastin similarily upregulated the expression of M2 macrophage markers in PBMC-derived macrophages.4.Erastin triggered a significant STAT3 phosphorylation in macrophages.5.While stattic successfully inhibited the activation of STAT3,it also eliminated erastin-induced M2 polarization of macrophages.6.Conditioned medium from macrophages treated with erastin could promote migration and invasion of EOC cells.Conclusion: Macrophages are not sensitive to erastin-induced ferroptosis.Erastin can enhance the invasion and migration of EOC cells by promoting M2 polarization of macrophages at least partially through the STAT3 signaling pathway.