Patients with intractable epilepsy (IE) exhibit an increased risk of premature death, psychosocial dysfunction and decreasing quality of life. The present study aimed to investigate the alteration in the expression of Rho7 in brain tissue from patients with IE, and to examine the association between Rho7 protein expression and IE. Temporal lobe samples were collected from the temporal lobes of 33 patients with IE patients and 10 age‑ and gender‑matched histologically healthy controls. Immunohistochemical staining was conducted to assess the number of Rho7‑positive cells. In addition, double‑label immunofluorescent staining was performed to examine the cellular localization of Rho7. The protein expression of Rho7 was examined using western blotting. Marked immunoreactivity for Rho7 was detected in the IE group, while faint and scattered immunoreactive staining was observed in the control group. The count of Rho7 positive cells in the IE patients was significantly increased compared with the control subjects (23.47±3.9% vs. 12.09±1.05%; P<0.01). Double‑label immunofluorescent staining indicated that Rho7 was primarily expressed in the cell membrane and cytoplasm, and colocalized with neuron‑specific enolase. Western blot analysis demonstrated that the expression of Rho7 in the IE group was significantly increased compared with the control group (0.41±0.031 vs. 0.25±0.025; P<0.01). The results of the present study demonstrated that upregulation of Rho7 immunoreactivity occurs in the brains of patients with IE, suggesting that Rho7 may be associated with the progression of IE or act as a potential treatment target.