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. 2020 Jun 24;8(1):e000285. doi: 10.1136/jitc-2019-000285

Figure 5.

Figure 5

IL-6 downregulates protein tyrosine phosphatase, receptor type O (PTPRO) in hepatocellular carcinoma (HCC) monocytes by upregulating miR-25–3 p via STAT3/c-MYC. (A, B) Heatmap and volcano plot indicating significantly expressed miRNA within monocytes in patients with HCC compared with the healthy controls. (C) Anti-CD14 magnetic beads were used to isolate human monocytes from 165 patients with HCC and 155 healthy controls. Transcription of miR-25–3 p was detected using real-time PCR and compared between the two groups. **p<0.01, Student’s t-test. (D, E) The linear correlations between expression of miR-25–3 p and PTPRO or PD-L1 were analyzed in HCC monocytes. (F) The potential binding site and mutation in the 3′ untranslated region (UTR) are indicated in the schematic figure. A luciferase reporter gene assay was carried out to test the promoter activity of PTPRO regulated by miR-25–3 p on the 3′UTR of PTPRO. (G, H) Linear correlations between serum IL-6 and monocyte PTPRO, and serum IL-6 and monocyte miR-25–3 p were analyzed in patients with HCC. (I) THP-1–derived and U937-derived macrophages were treated as indicated, and expression of miR-25–3 p was determined by real-time PCR. **p<0.01, Student’s t-test, compared with control group. (J) Cell signaling was analyzed by western blotting. Each experiment was performed in triplicate. Data are presented as the mean±SEM and were analyzed with Student’s t-test (**p<0.01).