The unsatisfactory clinical efficacy of dendritic cell (DC)-based cancer vaccines prepared by conventional methods is partly due to their insufficient capacity for migration. Our previous study showed that exposure to low-dose radiation (LDR) at a dose of 0.2 Gy promoted DC migration in vitro. The present study further investigates whether exposure to LDR at a dose of 0.2 Gy during the DC vaccine preparation could increase the antitumor effect of DC vaccines derived from mouse bone marrow. Our results showed that the migratory capacities of DCs were significantly increased after exposure to LDR. Furthermore, exposure to LDR resulted in an increased ability of DCs to induce T-cell proliferation, and the cytotoxic effect of cytotoxic T lymphocytes (CTLs) primed by the DCs exposed to LDR was significantly enhanced. An in vivo study using a mouse transplanted tumor model showed that subcutaneous injections of a DC vaccine exposed to LDR led to an increased mouse survival rate, infiltration of CTLs into tumor tissue, and apoptosis of tumor cells, which were accompanied by significant upregulation of serum interferon γ and interleukin 12. These results indicate that exposing DCs to LDR during the DC vaccine preparation is an effective approach to enhance its antitumor effect.