Fig. 5: USP22 promotes HCC-derived cell growth/invasion/Vascular Mimicry (VM) formation and angiogenesis.

A Knockdown of USP22 inhibited HCCLM3 proliferation. The absorbance of HCCLM3 cells carrying shCtrl or shUSP22 added MTS reagent in indicated time was measured at 490 nm. B, C Effects of USP22 on the cell growth in HCCLM3 cells were photographed in the colony formation assay. VEGFA-165 stands for recombinant human VEGFA protein. D The morphology of HCCLM3 cells changed with shUSP22 was photographed under microscope. E The left panel shows tube formation pictures of the HCCLM3 cells. Scale bars, 200 μm. The right panel is a quantification plot of the number of tubes in each visual field in the left. F, G Effect of USP22 on cell migration and invasion in Huh7 cells. Cells were planted in the transwell chamber coated with Matrigel (G) or without Matrigel (F) for 24 h. Scale bars, 250 μm. H ELISA assay was used to verify the effect of USP22 on VEGFA secretion from Huh7 cells. I Tube formation of HUVEC cells were photographed under microscope. Conditioned medium collected from shCtrl or shUSP22 Huh7 cells were applied to culture HUVEC cells on the Matrigel for 24 h. J Colony formation experiments were used to determine the effect of USP22 on the cell growth of HUVEC cells. Scale bars, 250 μm. Histogram data were shown as mean ± SD, P < 0.05 is considered statistically significant.