Fig. 7.

Exosomal miR‐223 promotes IL‐6 expression in monocyte/macrophage. (A) The proportion of immune cells was analyzed by CIBERSORT software based on the RNAseq data in TCGA cohort. (B) Value of IL‐6 and IL‐6R expression in miR‐223‐high or miR‐223‐low group in TCGA cohort. (C) IL‐6 level was determined in miR‐223‐OV or control cells by using qPCR. (D) IL‐6 level was determined by ELISA in SiHa culture, THP‐1 culture, or coculture as indicated. Data represent one experiment out of n = 3 performed in triplicates. (E, F) IL‐6 expression was examined by ELISA or qPCR in exosome‐treated macrophage derived from THP‐1 (10 μg·mL⁻¹ PMA for 24 h). Induced macrophage was treated with exosomes for 24 h, and LPS (100 ng·mL⁻¹) was added into the culture for another 24 h. Then, the RNA extracted from treated cells was reversely transcribed into cDNA for examining gene expression by using specific primers. (G, H) Phosphorylated STAT3 was examined by western blot assay using anti‐STAT3 (Try 705)‐specific antibody in SiHa cells which were cocultured with or without THP‐1 or U937 cells. Blots represent one of three independent experiments. (I) miR‐223 expression was examined by qPCR in IL‐6‐neutralizing antibody (50 ng·mL⁻¹)‐treated SiHa cells which were replaced with condition medium from THP‐1 culture simultaneously. (J) qPCR analysis of miR‐223 level in SiHa treated with or without anti‐IL‐6 antibody (50 ng·mL⁻¹) for 24 h (left), or in SiHa cells cocultured with or without THP‐1 which treated with or without anti‐IL‐6 antibody (right). Data were shown as mean ± SD. (K) IL‐6 was determined by qPCR in SiHa which was cocultured with THP‐1 treated with or without exosome inhibitor (GW4869 10 μm). (L) miR‐223 expression was examined in THP‐1, which was cocultured with or without SiHa, treated with or without exosome inhibitor. (M) Representative images of tumors from each group. (N) Quantitative analysis of the tumor weights in (M) *P < 0.05, **P < 0.01, ***P < 0.001.