Comparative evaluation of junctional adhesion molecule 3 and paired box gene 1 methylation, cytology, E6/E7 messenger RNA, and high-risk human papillomavirus testing for detecting precancerous cervical lesions

Objectives: This study evaluated paired box gene 1 ( PAX1 ) and junctional adhesion molecule 3 ( JAM3 ) methylation as biomarkers for detecting cervical precancerous lesions, comparing their performance with cytology, E6/E7 messenger RNA (mRNA) testing, and high-risk human papillomavirus (hrHPV) detection. Material and Methods: This retrospective study included 288 patients with abnormal human papillomavirus (HPV)-DNA or cytological findings warranting colposcopy who were enrolled between February 2023 and February 2024. Demographic data, cytology, HPV-DNA, E6/E7 mRNA, and PAX1/JAM3 methylation results were analyzed. Optimal methylation thresholds were identified using Youden’s index. Results: Methylation rates of PAX1 and JAM3 correlated with lesion severity: The combined positive rate of PAX1 and JAM3 methylation (positive for either marker) increased progressively from normal tissues (11.6%) to low-grade squamous intraepithelial lesion (LSIL) (27.8%), high-grade squamous intraepithelial lesion (HSIL) (79.2%), and cancer (100%). For HSIL + detection, PAX1/JAM3 methylation showed high sensitivity (82.0%), specificity (83.3%), and an area under the curve (AUC) of 0.826. Comparatively, AUCs for cytology, HPV 16/18, hrHPV, and E6/E7 testing ranged from 0.578 to 0.768. Combining PAX1/JAM3 with these methods improved diagnostic performance, with AUCs up to 0.831. Conclusion: PAX1 and JAM3 methylation testing demonstrates robust diagnostic potential for cervical precancerous lesions and complements existing screening methods.