[Abstract] Objective To investigate the expression and effect of Livin gene silencing by siRNA on growth and apoptosis in U251 cells. Methods The targeting Livin siRNA was prepared and transfected into U251 cells. The gene silencing efficiency was monitored by RT-PCR and Western blot analysis. The proliferation and apoptosis of U251 cells were detected by MTT assays and flow cytometry. The tumor volume and mean life span of tumor bearing mice were also observed. Results Recombinant adenovirus could silence Livin gene at the level of mRNA and protein, inhibited proliferation and enhanced apoptosis of U251 cells. In addition, Recombinant adenovirus could also inhibit the tumor growth and prolong the lifespan of tumor bearing mice. Conclusion The recombinant adenovirus Livin siRNA effectively silenced the expression of Livin gene, inhibited proliferation and enhanced apoptosis which provided a promising therapeutical strategy for glioma.