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Research Article: Development and validation of hypermethylated gene markers in cervical cytological samples for detecting endometrial cancer (EndoMethy-I trial)

Date Published: 2026-06-19

Abstract:
To explore hypermethylated gene markers in cervical scraped cells that may be associated with endometrial cancer and validate their diagnostic role in endometrial cancer. In an exploratory cohort consisting of 40 paired endometrial tissue and cervical scraping samples, high-performance methylation-targeted genes associated with endometrial cancer were identified. In training and validation sets with 347 and 149 participants, respectively, methylated markers from cervical cytology together with other epidemiological and clinical parameters were assessed to determine their accuracy in detecting endometrial cancer. A decision tree was constructed using methylation markers, bleeding symptoms and endometrial thickness on transvaginal ultrasound (TVS). In this exploratory study, eleven genes highly related to endometrial cancer were evaluated using a methylation array. In the training cohort, the highest AUC values for detecting endometrial cancer were 0.93, 0.91, and 0.89 for hypermethylated CDO1 , NEFM , and CELF4 , respectively. In the validation set, combining methylated CDO1 , CELF4 , and NEFM achieved a sensitivity of 94.6% (95% confidence interval 85.1-98.9) and a specificity of 92.8% (89.3-95.5) for detecting endometrial cancer. Integration of the endometrial thickness of TVS slightly improved the diagnostic specificity with very low sensitivity. Cervical cytological DNA methylation assays of CDO1 , CELF4 and NEFM provide a reliable and safe strategy for detecting endometrial cancer and are superior to other noninvasive evaluation methods or parameters.

Introduction:
Endometrial cancer, the most common gynecological cancer, is increasing in incidence worldwide, especially in high-income countries and cities ( 1 ). Increased risk of endometrial cancer is associated with a history of unopposed estrogen therapy, obesity, diabetes, and Lynch syndrome ( 2 – 5 ). Endometrial cancer is predominantly identified in women who present with abnormal uterine bleeding (AUB) or postmenopausal bleeding. Nonetheless, after invasive procedures and evaluations, only 0.33% of AUB patients and 9%…

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