The eukaryotic expression vector of human arresten gene was constructed and its secretive expression in human embryonic kidney (HEK 293) cells was detected.Human arresten gene was amplified from recombinant plasmid pGEM-Arr by polymerase chain reaction (PCR),and then digested with restriction endonucleases BamH I and EcoR I. The target fragment was inserted into the BamH I and EcoR I restriction sites of eukaryotic expression vector pSecTag2 to construct pST-AT. Restriction analysis and DNA sequencing indicated that the arresten gene was successfully inserted into pSecTag2. The recombinant plasmid was subsequently transfected into HEK 293 cells with LipofectAMINETM2000 Reagent,and the expression of the target gene was detected. Reverse transcription PCR (RT-PCR) revealed that the mRNA of the target gene was transcribed in the transfected HEK 293 cells. Weste blot analysis verified that the recombinant protein in supematants was correct. The supeatants of transfected cells were prepared, and 3-(4,5)-dimethylthiazol (-2-yl)-3,5-di-phenyltetrazoliumbromide (MTT) assay was carried out to assess their effects on the proliferation of human umbilical vein endothelial cells, which showed that the recombinant protein could significantly suppress the proliferation of human umbilical vein endothelial cells in vitro. These results provided a solid foundation to explore the usage ofarresten in tumor anti-angiogenic gene therapy.