Hepatocellular carcinoma-derived exosomal miRNA-761 regulates the tumor microenvironment by targeting the SOCS2/JAK2/STAT3 pathway

Xiao-hu Zhou, Hao Xu, Chang Xu, Ying-cai Yan, Lin-shi Zhang, Qiang Sun, Wei-lin Wang, Yan-jun Shi

Figure 3. Exosomal miR-761 induced the transformation of cancer-associated fibroblasts via the SOCS2/JAK2/STAT3 signaling pathway. A: the predicted miR-761 binding site within the SOCS2 3’-UTR and its mutated version; B: luciferase assay performed in WS1 cells after cotransfection with miR-761 or NC mimics and reporter vectors carrying wild-type or mutant SOCS2 3’-UTR; C: SOCS2 expression after transfection with NC or miR-761 mimics detected by quantitative reverse transcriptase-polymerase chain reaction; D: Western blotting to determine SOCS2 expression and JAK2/STAT3 signaling pathway; E: morphology of WS1 cells; F: representative images of migration assay; G: WS1 cell proliferation detection using CCK-8 assay; H: Western blotting detection of SOCS2, JAK2, STAT3, and MMPs expression. WS1 cells were treated with LM3-secreted exosomes and JAK2 inhibitor (AZD1480), or STAT3 inhibitor (C188-9). SOCS2: suppressor of cytokine signaling 2; FAP: fibroblast activation protein; α-SMA: α-smooth muscle actin; MMP-2: matrix metalloproteinase-2; MMP-3: matrix metalloproteinase-3; MMP-9: matrix metalloproteinase-9; STAT3: signal transducer and activator of transcription 3; JAK2: Janus kinase 2; GAPDH: glyceraldehyde-3-phosphate dehydrogenase. Compared with negative control grouip, *P<0.01, **P<0.001; compared with negative control grouip, #P<0.05, ##P<0.01.