miR-3682-3p directly targets FOXO3 and stimulates tumor stemness in hepatocellular carcinoma via a positive feedback loop involving FOXO3/PI3K/AKT/c-Myc

Figure 2 miR-3682-3p promotes the stem cell properties of hepatocellular carcinoma cells via the PI3K/AKT/c-Myc signaling pathway. A: Representative images of formed hepatospheres (× 100 magnification); scale bar: 200 μm; B: Flow cytometric analysis of the proportion of side population cells; C: Immunofluorescence staining was performed to assess the expression levels of CD44 and CD133 in hepatocellular carcinoma cells (scale bar: 5 μm); D and E: RT-qPCR analysis of SOX2, OCT4, CD133, and CD44 expression; F: Gene set enrichment analysis showing that miR-3682-3p regulates the PI3K/AKT signaling axis; G: Western blotting analysis of PI3K/AKT/c-Myc signaling- and stemness-related protein expression levels. ^bP < 0.01; ^cP < 0.001. Experiments were repeated three times. NC: Negative control.