Ovarian cancer remains a significant contributor to female mortality worldwide. Epigenetic alterations, which emerge during carcinogenesis, serve as potential biomarkers for cancer progression. Diabetes-related ankyrin repeat protein (DARP), also known as ankyrin repeat domain-containing protein 23, is a member of the muscle ankyrin repeat protein family and is encoded on chromosome 2q11.2. Aberrant methylation of the DARP gene promoter has been reported in various malignancies, including ovarian cancer. This study aimed to evaluate and compare the methylation status of the DARP gene promoter in women diagnosed with epithelial ovarian cancer to a control group consisting of individuals with benign ovarian tumors. A total of 155 female participants were enrolled in the study, comprising 98 patients with epithelial ovarian cancer and 57 controls with benign ovarian tumors. DNA was extracted from the formalin-fixed paraffin-embedded (FFPE) tissue samples of the participants. The methylation levels of CpG sites within the DARP gene promoter were quantitatively analyzed using pyrosequencing. The methylation levels at specific CpG sites were significantly elevated in women with epithelial ovarian cancer compared to the control group. Additionally, the mean methylation level was significantly higher in the ovarian cancer group compared to the controls (p < 0.001). The findings suggest that methylation of the DARP gene promoter may be relevant in the pathogenesis of ovarian cancer and serve as a predictive marker for disease progression and therapeutic decision-making.