By using overexpression and downregulation of HCC cell lines of HOXB5, we demonstrated that ectopic expression of HOXB5 in HepG2 cells promoted the capacity of cell growth and clonogenicity, while the inhibition of HOXB5 in Hep3B cells decreased the proliferation and clonogenicity potential in vitro by CCK8 and colony-forming assays. This evidence concerns the gene HOXB5 and hepatocellular carcinoma.