PURPOSE: To assess the degree and clinical impact of location error of the dens on the X-axis during radiotherapy to brain and head and neck tumors. MATERIALS AND METHODS: Twenty-one patients with brain tumors or head and neck tumors who received three-dimensional conformal radiation therapy or intensity-modulated radiation therapy from January 2009 to June 2010 were included in this study. In comparison two-dimensional verification portal images with initial simulation images, location error of the nasal septum and the dens on the X-axis was measured. The effect of set-up errors of the dens was simulated in the planning system and analyzed with physical dose parameters. RESULTS: A total of 402 portal images were reviewed. The mean location error at the nasal septum was 0.16 mm and at the dens was 0.33 mm (absolute value). Location errors of more than 3 mm were recorded in 43 cases (10.7%) at the nasal septum, compared to 133 cases (33.1%) at the dens. There was no case with a location error more than 5 mm at the nasal septum, compared to 11 cases (2.7%) at the dens. In a dosimetric simulation, a location error more than 5 mm at the dens could induce a reduction in the clinical target volume 1 coverage (V95: 100%-->87.2%) and overdosing to a critical normal organ (Spinal cord V45: <0.1%-->12.6%). CONCLUSION: In both brain and head and neck radiotherapy, a relatively larger set-up error was detected at the dens than the nasal septum when using an electronic portal imaging device. Consideration of the location error of the dens is necessary at the time of the precise radiation beam delivery in two-dimensional verification systems.