TRIM25 Promotes the Proliferation Migration and Invasion of Esophageal Squamous Cell Carcinoma Cells through EZH2-Mediated Macrophage M2 Polarization
Objective:To explore the roles of TRIM25 and EZH2 in infiltrating macrophages in esopha-geal squamous cell carcinoma(ESCC)and their potential mechanisms of action.Methods:THP-1 cells were induced to M0 macrophages using phorbol ester,co-cultured with KYSE510 cells after transfection,and the collected co-culture macrophages were divided into Ctrl group,sh-NC group,sh-TRIM25 group,sh-NC+oe-NC group,sh-TRIM25+oe-NC group,sh-NC+oe-EZH2 group,and sh-TRIM25+oe-EZH2 group.The co-culture supernatant was added to KYSE510 cells,and they were divided into Ctrl group,TAM group,sh-NC +TAM group,sh-TRIM25+TAM group,sh-NC+oe-NC+TAM group,sh-TRIM25+oe-NC+TAM group,sh-NC+oe-EZH2+TAM group,and sh-TRIM25+oe-EZH2+TAM group.qPCR and WB were used to detect the expression of TRIM25,EZH2,and Arg-1 in cells.Protein synthesis inhibition experiments were used to test the protein stability of EZH2.FCM was used to detect the proportion of F4/80+CD206+ macrophages.CCK-8,colony formation assay,and Transwell assay were used to evaluate cell proliferation,migration,and inva-sion.Results:The mRNA and protein expression of TRIM25 and EZH2 in KYSE510 cells were higher than in human esophageal squamous epithelial cells(HET-1A)(P<0.01).Compared with the sh-NC group and sh-NC+oe-NC group,the proportion of F4/80+CD206+ macrophages and the expression of Arg-1 protein in the sh-TRIM25 group and sh-TRIM25+oe-NC group decreased(P<0.05),while it increased in the sh-NC +oe-EZH2 group(P<0.05).Compared with the sh-NC+TAM group and sh-NC+oe-NC+TAM group,the sh-TRIM25+TAM group and sh-TRIM25+oe-NC+TAM group showed decreased KYSE510 cell viability,col-ony formation,migration,and invasion(P<0.05),while the sh-NC+oe-EZH2+TAM group showed an in-crease(P<0.05).Knockdown of TRIM25 could inhibit the protein half-life of EZH2.Overexpression of EZH2 partially reversed the effects of sh-TRIM25 on macrophages and KYSE510 cells.Conclusion:TRIM25 induces M2 polarization of macrophages by promoting the stability of EZH2 protein,thereby promoting the pro-liferation,migration,and invasion of esophageal squamous cell carcinoma cells.
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