Chemokines have been shown to play a critical role in tumor development and progression. However, little is known about the function and molecular mechanisms of CXCR6 in multiple malignancies. In the present study, we aimed to investigate the role of CXCR6 in human hepatocellular carcinoma (HCC). The expression of CXCR6 was examined by immunohistochemical assay using a tissue microarray procedure. A loss-of-function experiment was performed to explore the effects of lentivirus-mediated CXCR6 shRNA (shCXCR6) on cell proliferation and invasive potential by MTT and Transwell assays in HCC cell line (SMMC-7721). It was found that the expression of CXCR6 protein was significantly increased in HCC tissues compared with that in adjacent non-cancerous tissues (ANCT) (63.04% vs 36.96%, P=0.019), and correlated with the lymph-vascular space invasion in HCC patients (P=0.038). Knockdown of CXCR6 repressed cell proliferation and invasion of HCC cells followed by the down-regulation of vascular endothelial growth factor (VEGF). Taken together, our findings show that high expression of CXCR6 is positively associated with distant invasion of HCC patients, and blockade of CXCR6 signaling suppresses the growth and invasion of HCC cells through inhibition of the VEGF expression, suggesting that CXCR6 may represent a promising therapeutic target for the treatment of HCC.