Objective:To construct the recombinant lentiviral vector containing LMP2A gene and measure the expression level of LMP2A in human carcinoma cell line of CNE1, as well as the effects of LMP2A on proliferation, migration and invasion on human carcinoma cell line of CNE1. Methods: The EB virus LMP2A fragment was amplified by PCR and subcloned into the lentiviral vetor plv by recombinant DNA technology. The resultant lentivirus was confirmed by PCR, restriction enzyme digestion and DNA sequencing. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pLMP2A and packaging plasmid pdelta-8.91 and pVSVG. CNE1 cells were infected by plv-LMP2A, and the expression of LMP2A in CNE1 cells was confirmed by RT-PCR, immunofluorescence and Western blot by screening of monoclonal. CCK8 assay, Transwell invasion assay and Wound-healing assay were performed to determine the effects of LMP2A expression on the proliferation, invasion and migration. Results: The recombinant lentiviral vector carried the LMP2A gene was successfully constructed. Results of RT-PCR, immunofluorescence and western blot indicated that CNE1 transgenetic cells could high express EBV-LMP2A. The LMP2A gene-transfected carcinoma cells grew vigorously and rapidly and up-regulated the ability of migration and invasion. Conclusion: The recombinant lentiviral vector pLMP2A was successfully constructed, and could be used to transfect human carcinoma cell line of CNE1. LMP2A could be highly expressed and can significantly promote CNE1 proliferation, migration and invasion, which laid a foundation of study on the biological function of Epstein-Barr virus and its potential role in gene therapy of tumors. [Key words] Epstein-Barr virus; latent membrane protein 2A; nasopharyngeal carcinoma; lentiviral vector