Objective To explore the effects of hsa-miR-96 to A549. Method A549 were transfected with miR-96 inhibitor/mimics and shRNA-mTOR. The proliferation were detected by MTT,the expression of p21, cyclin D1,p-4E-BP,S6K were detected via RT-PCR and WB. The luciferase assay were used to anaylse the target of miR-96. Result With or without metformin treated,up-regulated the expression of miR-96 could inhibit A549 proliferation,increase p21 expression and decrease cyclin D1 expression instead. The results may related with G1 arrest which induced by miR-96 up-regulation. The ratio of firefly fluorescence value/Renilla fluorescence value was lower than that in wild type NC or mutant group. Conclusion miR-96 target with mTOR to inhibit the growth of A549.