Objective: To study the expression and significance of apoA-I binding protein (AIBP) in hepatocellular carcinoma (HCC) cells. Methods: The mRNA and protein expressions in normal hepatic L02 cells, AFP-positive HCC HepG2 and Hep3B cells, and AFP-negative HCC SMMC7721 cells were measured by RT-PCR and Western blot, respectively. The recombinant vector pcDNA3.1-AFP-AIBP-yCD/TK that contained CD and TK double suicide genes driven by AFP promoter with AIBP gene overexpression was constructed, and then, the vectors were transfected into the Hep3B and SMMC7721 cells. After transfection, proliferation in both types of cells was determined by MTT assay, and the mRNA and protein expressions of AIBP, vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR-2) and matrix metallopeptidase 9 (MMP-9) were detected by RT-PCR and Western blot, respectively. Results: Both the AIBP mRNA and protein presented high expression in the normal hepatic cells, while those were significantly down-regulated in all the tested HCC cells with the down-regulation more evident in Hep3B and SMMC7721 cells. The recombinant plasmid pcDNA3.1-AFP-AIBP-yCD/TK was successfully constructed and transfected into Hep3B and SMMC7721 cells. After transfection, the proliferation of AFP-positive HepG2 cells was significantly inhibited, but the proliferation of AFP-negative SMMC7721 cells was not affected; in both types of cells, the mRNA and protein expressions of AIBP were remarkably increased, while the mRNA and protein expressions of VEGFR-2, VEGF and MMP-9 were reduced. Conclusion: AIBP expression is down-regulated in HCC cells, which is associated with the invasion and metastatic ability but not with the proliferative ability of HCC cells. Meanwhile, the combination gene vector pcDNA3.1-AFP-AIBP-yCD/TK has been successfully constructed.