The present study aimed to investigate the influence of COL8A1 expression on cell invasiveness, drug sensitivity and tumorigenicity of hepatocellular carcinoma Hepa1-6 cells with low metastatic potential. COL8A1-1-pEGFP-N2 and pEGFP-N2 were transfected into experimental and control group cells. The COL8A1 expression in transfected Hepa1-6 cells was analyzed with RT-PCR and western blot analysis. The invasive potential of transfected Hepa1-6 cells was tested in invasion experiments in vitro and the tumorigenic ability of the transfected Hepa1-6 cells was tested in mouse tumors in vivo. Hepa1-6 cell proliferation and D-limonene sensitivity was analyzed using the MTT method. Expression of COL8A1 in the Hepa1-6/COL8A1 group showed a significant increase when compared with the untransfected cells of the Hepa1-6 control group and empty-plasmid transfected cells from the Hepa1-6/mock control group. Enhanced COL8A1 expression increased cell proliferation and matrix adhesion ability via invasion and tumorigenesis in vivo while the sensitivity to D-limonene was concurrently inhibited. The expression of COL8A1 in hepatocarcinoma cells was correlated with increased tumor cell proliferation, invasion, in vivo tumorigenicity and reduced antitumor drug sensitivity, and may provide novel targets for tumor therapy.