Abstract：Objective: To explore the roles of TRIM25 and EZH2 in infiltrating macrophages in esophageal 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 invasion. 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, colony formation, migration, and invasion (P<0.05), while the sh-NC+oe-EZH2+TAM group showed an increase (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 proliferation, migration, and invasion of esophageal squamous cell carcinoma cells.
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2024, Vol. 30 
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