Abstract: ObjectiveTo investigate the effects of adenovirus-mediated shRNA targeting c-jun N-terminal kinase 1(JNK1) in U87MG human glioblastoma cells, so as to reveal the relationship between JNK1 and cell proliferation in brain tumor. Methods shRNA sequence against JNK1 and nonsense sense oligonucleotides were designed; after producing recombinant adenovirus, Q-PCR and Western blot were used to test knockdown effect in U87MG cells; BrdU incorporation was used to observe cell proliferation. Results Recombinant vector was confirmed by restriction endonuclease analysis and DNA sequencing; Q-PCR and Western blot results indicated the expression of JNK1 was significantly decreased compared with the other control groups. BrdU incorporation revealed cell proliferation rate was markedly inhibited in JNK1 RNAi group in comparison with control groups. ConclusionRecombinant adenovirus expressing shRNA targeting JNK1 was constructed successfully, and cell proliferation was significantly inhibited after JNK1 knockdown in U87MG cells; it paves the way for further research on JNK pathway involving in development of brain tumor.