1Background & Objective
hrHPV
high Se, low Sp
TCT
subjective
Methy
early event
- Screening challenge: hrHPV high sensitivity but low specificity; cytology depends on reader experience.
- Epigenetics: DNA methylation is an early event; PAX1/JAM3 hypermethylation tracks lesion progression.
- Aim: evaluate PAX1/JAM3 methylation in exfoliated cells for predicting CIN2+.
2Study Design
144
subjects
2
methylation genes
4
test methods
- Cohort: Jul 2023 – Dec 2024, Changsha Maternal & Child Health Hospital; 144 subjects.
- Tests: HPV genotyping + TCT + PAX1/JAM3 methylation + colposcopy pathology for all.
- Endpoint: predictive performance for high-grade CIN (CIN2+).
- Groups: CIN2+ vs CIN1 vs chronic cervicitis comparison for methylation positivity.
68.8%
PAX1+ in CIN2+
56.3%
JAM3+ in CIN2+
1.2–6.4%
controls
4
assays per subject
18 mo
enrollment window
CIN2+
primary endpoint
- Sampling: non-invasive cervical exfoliated cells — same sample used for cytology and methylation.
- Combined strategy: methylation readout integrated with HPV genotyping for risk stratification.
- Statistics: ROC/AUC for combined model; PPV/NPV; OR with 95% CI; P<0.05 significant.
ZHOU M et al. · J Hunan Normal Univ (Med Sci) 2025;22(6):123-128 · Changsha MCH.
Methylation panel: PAX1 & JAM3 (dual-gene) on cervical exfoliated cells; combined with HPV genotyping.
Reference: colposcopy-directed pathology (CIN2+ as endpoint).
设计要点：144 例均完成 HPV 分型、TCT、PAX1/JAM3 甲基化与阴道镜病理；终点为 CIN2+ 预测效能；甲基化阳性率随病变加重呈剂量-反应。
核心结果：CIN2+ 中 PAX1、JAM3 甲基化阳性率分别为 68.8% 与 56.3%，对照组仅 1.2%–6.4%（P<0.001），呈剂量-反应关系。
3Methylation Positivity by Group
68.8%
CIN2+ PAX1
56.3%
CIN2+ JAM3
6.4%
CIN1 PAX1
1.2%
cervicitis
- Dose–response: positivity rises with lesion severity (cervicitis 1.2% → CIN2+ 68.8%, P<0.001).
- Early signal: CIN1 PAX1+ 6.4% — low, consistent with early-stage biology.
- Discrimination: methylation clearly separates CIN2+ from controls.
4Combined Prediction Strategy
| Strategy | Metric | Result |
|---|---|---|
| Methylation + HPV genotyping | AUC | 0.8789 |
| HPV alone | F1 | 59.1% |
| HPV16/18 + any methylation+ | PPV | 96.9% |
| hrHPV+ & methylation+ | OR (95% CI) | 6.32 (3.45–11.28) |
- Combined gain: dual-gene methylation + HPV typing significantly outperforms HPV alone (F1 59.1%, P<0.01).
- Highest PPV: HPV16/18 + any methylation+ → CIN2+ PPV 96.9% — practical for triage of uncertain cytology.
- Risk link: hrHPV infection with methylation+ → CIN2+ OR 6.32.
5Risk Stratification & Value
- High-risk flag: HPV16/18 + methylation+ → CIN2+ PPV up to 96.9% — prioritize colposcopy.
- Low-risk triage: methylation-negative hrHPV carriers have markedly lower CIN2+ risk — fewer unnecessary referrals.
- Uncertain cytology: ASC-US/LSIL cases can be objectively stratified by methylation status.
- Objective + accessible: exfoliated-cell sampling is convenient for outpatient opportunistic screening.
- Limit: preliminary, limited sample; needs larger prospective validation.
59.1%
HPV-alone F1
↑
combined gain
0.8789
combined AUC
- F1 improvement: combined model significantly raises F1 over HPV alone (P<0.01).
- Patient benefit: fewer unnecessary colposcopies and less anxiety for low-risk carriers.
Methylation complements HPV (viral infection) by measuring host malignant transformation.