To investigate the expression of FUT8, STX4, and calpain2 in ovarian tumor tissues and their association with clinicopathological characteristics.This retrospective study analyzed the expression of FUT8, STX4, and calpain2 in ovarian tumor samples from patients treated at Shanghai Pudong New Area People's Hospital between January 1, 2008, and December 31, 2017. The association between marker expression and histopathological features was evaluated. Binary logistic regression was used to assess the diagnostic value of these markers in malignant ovarian tumors.The expression of FUT8, STX4, and calpain2 was significantly higher in malignant ovarian tumors compared to benign tumors (all P < 0.05). A positive correlation was observed among the expressions of FUT8, STX4, and calpain2 (all P < 0.001). FUT8 expression was associated with the International Federation of Gynecology and Obstetrics (FIGO) stage (P = 0.013), STX4 expression was related to patient age (P = 0.012) and lymph node metastasis (P = 0.019). Patients with high co-expression of all three markers had a significantly higher likelihood of having malignant ovarian tumors compared to those with low expression of at least one marker (P = 0.002).Simultaneous detection of FUT8, STX4, and calpain2 expression in ovarian tumor tissues provides valuable diagnostic insights for malignant ovarian tumors.