Moreover, adherent foci formation in monolayer culture (P < 0.01, Fig. 2D) and non-adherent colony formation in soft agar (P < 0.05, Fig. 2E) assays also found the increased number of colonies of HepG2 and Huh7 cells after transfection of TROAP. Orthotopic liver injections of HCC cells in nude mice showed the larger tumor burden of mice injected with TROAP-overexpressed cells, indicating the tumor growth promotion of TROAP (P < 0.01, Fig. 2F). This evidence concerns the gene TROAP and hepatocellular carcinoma.