Association of PAX1/JAM3 gene methylation, HR-HPV, and TCT with high-grade cervical lesions in a high-risk cohort: a multinomial logistic regression analysis

To investigate the risk factors associated with pathological progression of cervical lesions, specifically focusing on high-risk human papillomavirus (HR-HPV) infection, ThinPrep Cytologic Test (TCT), and PAX1/JAM3 gene methylation in a high-risk colposcopy-referred cohort. A total of 409 patients who underwent colposcopy-directed biopsy at the First Hospital of Hebei Medical University between January, 2023 and April, 2025 due to abnormal TCT results, HR-HPV infection, or positive PAX1/JAM3 gene methylation were analyzed. The sensitivities of different methods were compared, and unordered multivariate logistic regression was used to analyze the risk factors for high-grade cervical lesions and pathological progression. Univariate analysis demonstrated that age, gravidity, age at first sexual intercourse, number of sexual partners, menopausal status, HR-HPV infection, TCT results, and PAX1/JAM3 methylation were significantly associated with pathological grade (all P < 0.05). In the multinomial regression analysis, compared to the inflammation group, both other high-risk HPV (OR = 3.930) and very-high-risk (16/18) HPV infections (OR = 3.725), as well as non-NILM TCT (OR = 2.435), were significantly associated with an increased risk of LSIL. For the progression to HSIL, very-high-risk HPV (OR = 6.801), non-NILM TCT (OR = 2.399), positive PAX1 methylation (OR = 4.145), and positive JAM3 methylation (OR = 8.215) were identified as strong independent risk factors. In a high-risk colposcopy-referred cohort, HR-HPV infection, non-NILM TCT results, and positive PAX1/JAM3 gene methylation are independently associated with the presence of high-grade cervical lesions. These biomarkers have the potential to enhance clinical risk stratification and inform decision-making in cervical cancer screening. However, their clinical utility and cost-effectiveness need to be further evaluated through larger, prospective studies to validate their role in routine clinical practice.