Fig. 7
GNB4 induces GC malignancy through the Hippo–YAP1 pathway in vitro and in vivo. A, B MKN45 (A) and AGS (B) cells were treated with different concentrations of verteporfin (VP; YAP1 inhibitor), and half-maximal inhibitory concentrations (IC50) of the VP were measured by CCK-8 assay after 24 h. C–F VP was added to MKN45 and AGS cells with or without GNB4 overexpression for 24 h at concentrations obtained through IC50 analysis. CCK-8 (C, D) assay detected the proliferative capacity of cells, and transwell assays (E, F) monitored the migration ability of cells. Representative images (scale bar: 100 μm; left panels E and F) and histogram show the relative number (with reference to negative control) of cells that passed through the chamber membrane in each group (right panels E and F). G–I Subcutaneous tumor models (n = 6 mice per group) were established using stable GNB4-overexpressed MKN45 cells combined with VP intraperitoneal injection (100 mg/kg, once daily, 7 times). Growth curves (G) and tumor weights (I) were analyzed. Mass images of tumors collected from mice are shown (H). J Western blot analysis of GNB4 protein expression in HGC27 cells transfected with lentivirus vector or overexpressing exogenous GNB4. K–O Mouse xenograft assays: HGC27 and MKN45 cells with or without GNB4 overexpression were injected in the tail vein of athymic nude mice, and 3 weeks after cell inoculation, the mice were treated with VP (100 mg/kg) daily for 7 days (n = 6 mice per group). Representative mass images (scale bars: 1 mm, K) of mouse liver and representative H&E images (scale bar: 50 μm; inset: 20 μm) of liver samples (HGC27 cells injected, L) and lung samples (MKN45 cells injected, O) from the indicated groups of nude mice. The number of metastatic nodules in mouse liver (M) and lung (N) was analyzed