Fig. 6: AZD7762, a small molecular inhibitor of VRK2, can reduces HCC growth.

a Molecular docking of AZD7762 with VRK2 (PDB:2V62). b IC50 assay demonstrated high inhibitory potency and selectivity of AZD7762 for VRK2 (n = 6 independent experiments). c Enzyme activity assay confirmed VRK2 selectivity (n = 6 independent experiments). d In vitro kinase assay showed AZD7762 inhibited MYC phosphorylation, proteins were harvested for western blotting with the indicated antibodies (n = 6 independent experiments). e CCK-8 assays in VRK2-knockout MHCC97H cells treated with DMSO/AZD7762 for 12 h (n = 6 independent experiments). f Representative images of colony formation and quantification (left/right panel) post-treatment (n = 6 independent experiments). g Representative EdU assay image of MHCC97H cells after VRK2 knockout, which were treated with DMSO or AZD7762 for 12 h and quantification (left/right panel) post-treatment (n = 6 independent experiments). h qRT-PCR analysis of PD-L1 expression, which was treated with DMSO or AZD7762 for 12 h (n = 3 independent experiments). i Western blot analysis of PD-L1 expression, which were treated with DMSO or AZD7762 for 12 h, cells were harvested for western blotting with the indicated antibodies (n = 3 independent experiments). j Flow cytometry analysis of PD-L1 expression, which was treated with DMSO or AZD7762 for 12 h (n = 3 independent experiments). k Schematic of DEN/CCL₄-induced HCC model in WT C57BL/6J mice (n = 7 mice in each group) and dosing strategy. l Liver tumor images: H&E, Oil Red, Ki67, PD-L1 and CD8 staining of isolated liver tissues. Gross image Scale bar: 1 cm, IHC image Scale bar: 200 μm. m Western blot analysis in mice HCC tissues. After prophylactic and therapeutic of AZD7762, cells were harvested for western blotting with the indicated antibodies (n = 3 independent experiments). n Tumor volume of the indicated models (n = 7 mice in each group). o Tumor numbers of indicated models (n = 7 mice in each group). p, q Serum AST/ALT levels in models (n = 7 mice in each group). r Body weight measured. After prophylactic and therapeutic of AZD7762 indicated models (n = 7 mice in each group). For Western blot experiments, GAPDH was used as a loading control (i, m). c, e, g, h, n–r P values were calculated using a student’s unpaired t-test (two-tailed). Data were presented as the mean ± SD. Source data are provided as a Source Data file.