Objective To investigate the effect of lentivirus-mediated shRNA targeting AKT1 gene on the sensitivity of gastric cancer xenografts to doxorubicin. Methods To establish the lentiviral vector of AKT1 RNA interference (RNAi), the vector of pGCSIL-shAKT1-GFP was infected into HEK293T cells, and meanwhile, the empty vector pGCSIL-shCON-GFP was assigned as a blank group, and then the viruses were harvested. BGC-823 gastric cancer cells were infected with the viruses. The protein expression of AKT1 was detected by Western blotting. The gastric cancer xenografts in nude mice were constructed using BGC-823 cells infected with the viruses, followed by the administration of doxorubicin. The size of tumor was evaluated and the growth curve of the tumor was drawn; HE staining was used to observe the pathological conditions of the xenografts; and the apoptosis of xenografts was examined by TUNEL assay. Results The recombinant plasmid of LV-shAKT1 was successfully constructed, and then transfected into HEK293T cells to produce high-titer lentivirus with a titer of 5×108 TU/mL. The expression of AKT1 protein in gastric cancer BGC-823 cells was significantly down-regulated after successfully infected with the LV-shAKT1. Nude mice xenografts experiment showed that AKT1 gene silencing inhibited the growth of tumor xenografts, and also enhanced the inhibitory effect of doxorubicin on the growth of tumor xenografts. TUNEL staining showed that AKT1 gene silencing promoted the apoptosis of tumor xenograft cells, and the effect was more obvious when combined with doxorubicin. Conclusion AKT1 gene silencing can enhance the sensitivity of gastric cancer xenografts to doxorubicin through promoting cell apoptosis.