Objective: To construct lentiviral vector carrying siRNA targeting Jagged 2 (JAG2) gene and observe the effects of its transfection on biological characteristics in human pancreatic cancer cells. Methods: Several lentiviral expression vectors carrying different siRNA fragments targeting JAG2 gene were constructed using gene recombination technology, and were transfected into pancreatic cancer cells primarily isolated from the human pancreatic cancer tissue specimens through enzymolysis approach. Then, the siRNA fragment showing strongest inhibitory effect on JAG2 gene was selected for testing the influences of its transfection on proliferation, apoptosis, cell cycle and invasion and metastasis abilities in human pancreatic cancer cells by MTT assay, flow cytometry and transwell chamber assay, respectively. Results: All the lentiviral expression vectors carrying different siRNA fragments showed inhibitory effects, to different extents, on mRNA and protein expressions of JAG2. In pancreatic cancer cells after transfection with the JAG2 siRNA, the proliferation was decreased, apoptosis and S-phase arrest were enhanced, and invasion and metastasis abilities were reduced significantly (all P<0.05). Conclusion: lentiviral vector bearing JAG2 siRNA are successfully constructed and their transfection can effectively moderate the malignant biological characteristics of human pancreatic cancer cells.