Findings from brain imaging studies investigating the neural underpinnings of antisocial behavior have implicated the prefrontal cortex in the regulation of aggressive reactions. However, relatively few studies have examined the role of the prefrontal cortex on aggression in an experimental way. This study examines whether upregulating the prefrontal cortex using repeated transcranial direct current stimulation (tDCS) reduces aggressive behavior. In a double-blind, parallel-group, randomized controlled trial, 88 healthy adults (42 males, 46 females) were assigned to one session of anodal tDCS over the ventromedial prefrontal cortex (n = 47) or sham stimulation (n = 41) per day for three consecutive days and assessed using a behavioral measure of aggression. Levels of aggressive responses post-intervention did not significantly differ between the active and sham stimulation groups. However, a significant interaction effect between the stimulation group and gender was observed, whereby males, but not females, exhibited reduced aggression after prefrontal stimulation. To the authors' knowledge, this is the first study to examine the effect of multi-session prefrontal tDCS on aggressive behavior in healthy adults. Results highlight that there are differences in responsivity to tDCS in modifying aggressive behavior.