Fig. 6

The mTORC1/STAT3/miR-130b-3p/MBNL1 signaling pathway exists in human cancer. A Expression analysis of miR-130b-3p in HNSCC and ANM tissues using TCGA. B GSEA analysis comparing the gene sets positively regulated by mTORC1 signaling in miR-130b-3p-high and miR-130b-3p-low HNSCC patients based on TCGA datasets. C miR-130b-3p levels were measured by qRT-PCR using the paired ANM tissues and HNSCC tissues (n = 60). D Representative IHC images of p-S6 staining from HNSCC tissues. Scale bar, 50 μm. E The correlation between p-S6 and miR-130b-3p expression of HNSCC tissues. F p-S6 and miR-130b-3p levels of the indicated cells were analyzed by qRT-PCR and western blot, respectively. G FaDu cells were treated with rapamycin (20 nM or 50 nM) for 24 h. H FaDu cells were infected with lentivirus expressing shRNAs targeting Raptor (shRaptor) or a control shRNA (shRNA). I FaDu cells were transduced with lentiviruses expressing sponge sequences of miR-130b-3p (miR-130b SP) or a control lentivirus. G-I Cell lysates were subjected to immunoblotting with the indicated antibodies (left panels), the expression of miR-130b-3p was detected by qRT-PCR (right panels). J and K The pro-angiogenic effect of miR-130b-3p was detected by tube formation (J, scale bar, 50 μm) and CAM assays (K). Representative images (left panels) and quantifications (right panels) are shown. (L-O) miR-130b SP-expressed FaDu cells and the control cells were inoculated subcutaneously into nude mice, and tumor growth was monitored. L Tumor pictures. Scale bar, 1 cm. M Tumor growth curves. N Tumor weight. O Representative HE and IHC staining of the indicated tumor tissues. Scale bar, 50 μm. Data indicate mean ± SD of 3–5 replicates. ** P < 0.01; *** P < 0.001; **** P < 0.0001; n.s indicates no significant difference