Additionally, we established two models of metastatic HCC, including tail vein injected models (SMMC7721 and HepG2) and an orthotopic model (HCCLM3), to investigate roles of CLU in HCC metastasis in vivo. Compared to mock cells, CLU knockdown resulted in significant decrease of metastatic foci in SMMC7721 cells (40.9 ± 6.0 vs. 20.9 ± 2.5, P = 0.0008, Figure 2G) and HCCLM3 cells (50.8 ± 5.6 vs. 20.6 ± 5.6, P = 0.0032, Figure 2H). This evidence concerns the gene CLU and hepatocellular carcinoma.