Tumor-derived exosomal miR-425-5p and miR-135b-3p enhance colorectal cancer progression through immune suppression and vascular permeability promotion

Figure 2 Exosomal miR-425-5p inhibition promotes macrophage M1-like polarization and proinflammatory T cell differentiation. A: Flow cytometry analysis of CD86⁺iNOS⁺ and CD163⁺CD206⁺ cells ratio in THP-1 cells cultured with exosomes isolated from HTC116 cells transfected with negative control and miR-425-5p inhibitor; B: Immunofluorescent staining of CD86 (red) and CD206 (green) in THP-1 cells with DAPI nuclei (blue); C: Real-time quantitative polymerase chain reaction analysis of cytokine expression in CD4⁺ T cells cultured with exosomes; D and E: Flow cytometry analysis of CD4⁺ T cell subsets. ^aP < 0.05, ^cP < 0.001 vs negative control-exosome (n = 3 per group). NC-exo: Negative control-exosome; CD4: T-cell surface glycoprotein CD4; CD25: Interleukin-2 receptor subunit alpha; CD86: T-lymphocyte activation antigen CD86; CD163: Scavenger receptor cysteine-rich type 1 protein M130; CD206: Macrophage mannose receptor 1; FoxP3: Forkhead box protein P3; IFN-γ: Interferon gamma; IL-1β: Interleukin 1 beta; IL-4: Interleukin 4; IL-6: Interleukin 6; iNOS: Inducible nitric oxide synthase; Th: T helper cells; TNF-α: Tumor necrotic factor alpha; Treg: Regulatory T cells.