Objective: To construct the recombinant lentiviral vector containing human annexin A2 (ANXA2) gene and measure the expression level of ANXA2 in hepatocellular cell line of Hepa 1-6. Methods:The ANXA2 fragment was amplified by PCR and subcloned into the lentiviral vector pWPT. The resultant lentivirus were confirmed by PCR, restriction enzyme digestion and DNA sequencing. Recombinant lentivirus (LV-ANXA2-Flag) were produced from 293T by transient calcium-phosphate co-transfection with pWPT-ANXA2-Flag, CMVR8.91and pCMV-VSV-G. Hepa 1-6 cells were infected by LV-ANXA2-Flag lentivirus, and the expression of ANXA2 was confirmed by RT-PCR and Western blotting. Cell cycles were measured via flow cytometry. Results:The recombinant lentiviral vector carried the ANXA2 gene was successfully constructed. RT-PCR and Western blot analysis revealed that the ANXA2 gene can be correct transcripted and translated in Hepa 1-6 cell which infected by LV-ANXA2-Flag. After infected,the Hepa 1-6 cells were higer than control cells in percentage of S phase cells. Conclusion: The recombinant lentiviral vector pWPT-ANXA2-Flag was constructed successfully, it could be used to transfect hepatocellular carcinoma cells. ANXA2 could be stablely expressed and promote the proliferation of Hepa 1-6 cell.