Abstract: Objective To clone the protein-coding region of paxillin （PXN） gene and construct its eukaryotic expression vector to establish hepatocellular carcinoma cell （HCC） line stably transfectd with PXN gene, aiming to evaluate the effect of PXN gene upon HCC. Methods Total RNAs of HCC were extracted by Trizol in vitro, and then reversely transcribed into cDNAs to amplify protein-coding region of PXN gene by PCR. The protein-coding region of PXN gene was inserted into eukaryotic expression vector of pEBFP-C1 to construct the recombinant plasmid pEBFP-PXN. Then, the pEBFP-PXN was stably transfected into Hep G2 cells by liposome, and the treated cells were screened with G418 to obtain stably transfected cell line which highly expressed paxillin. The cell growth rate was detected by real-time cellular analysis. The cell migration rate was determined by cell scratch wound assay. The expression levels of proteins were detected by Western blot. Results The protein-coding region of PXN gene was successfully cloned and the recombinant eukaryotic expression vector pEBFP-PXN was constructed. Monoclonal stably transfected HepG2 cells which highly expressed paxillin was obtained and seen in blue fluorescence. Western blot demonstrated that stably transfected cell line expressed blue fluorescent protein （BFP）-PXN. The expression of exogenous paxillin did not influence the growth rate of Hep G2 cells but inhibited the cell migration rate. Furthermore, the expression of epithelial marker protein E-Cadherin was up-regulated, whereas the expression levels of mesenchymal markers N-Cadherin and Vimentin were down-regulated in stably transfected Hep G2 cells. Conclusion PXN gene exerts no upon the growth rate, whereas suppresses the cell migration of Hep G2 cells, probably because paxillin can affect the expression of epithelial-mesenchymal transition markers.