Figure 10. Administration of Ad-miR-223 suppresses PD-1/PD-L1 axis and angiogenesis in inflammation-related orthotopic HCC model.

C57BL/6J mice were treated with CCl₄ plus Hepa1-6-implantation and were treated with Ad-miR-223 (n=8) or Ad-GPF (n=6). (A) Gross images of livers. (B) Tumor volume (mm³) in Ad-GFP and Ad-miR-223 treated-mice were analyzed. (C) miR-223 levels in tumor tissues of Ad-miR-223- or Ad-GFP-treated mice were measured. (D) Protein levels of HIF-1α, PCNA, phosphorylated VEGFR2, total VEGFR2 were determined by western blot analysis. (E) Quantitative analysis of HIF-1α, PCNA, total VEGFR2 and ratio of phosphorylated VEGFR2 to total VEGFR2 (p-VEGFR2/VEGFR2 ratio). (F) Upper panel: Representative images of H&E and immunostaining (CD31, Ki-67, PD-1, PD-L1) in consecutive liver sections from Ad-GFP and Ad-miR-223-treated groups are shown. Lower panel: quantification of CD31, Ki-67, PD-1, PD-L1 staining. Tumor region was surrounded by dashed white line. (T: tumor region). Values represent means ± SEM. *P< 0.05, **P< 0.01, ***P< 0.001.