Methylation-based biomarkers from thinprep cytology test samples for early detection of cervical cancer: application and clinical potential

Sep 22, 2026

Although HPV testing and Thin-Prep Cytology Test (TCT) are widely used for first-line cervical cancer screening, they face limitations. This study develops and validates methylation-based biomarkers from formalin-fixed paraffin-embedded (FFPE) tissue and TCT samples in a Chinese population to enhance early detection of cervical cancer. Differential methylation analysis and targeted methylation sequencing were used, followed by qMSP to build a diagnostic model and evaluate clinical performance in multi-group cohorts.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages11

Reading Time28 min

AccessFree to download

Background

Methods

Results

Conclusions

Keywords

Liu et al. Clinical Epigenetics (2026) 18:156 [https://doi.org/10.1186/s13148-026-02154-5](https://doi.org/10.1186/s13148-026-02154-5) Clinical Epigenetics RESEARCH Open Access Methylation-based biomarkers from thinprep cytology test samples for early detection of cervical cancer: application and clinical potential Zhihan Liu1†, Guorong Liu1†, Shuanping Liu2†, Xiuxiu Wang2, Yunshi Liang1, Yuanyuan Guo2, Baochen Du2, Nabo Miao1, Wenli Shi1, Qiuyun Zhu3, Liping Yu3, Cui Deng3, Guo Chen3, Lijuan Huang3, Lingling Zhan1,4, Xiangyu Zheng1,5, Jie Zhao2, Yongjun Li2*, Xiaoliang Han2*, Yang Cheng3* and Jing Yang1* Abstract Background Although human papillomavirus (HPV) testing and Thin-Prep Cytology Test (TCT) are currently adopted as the first-line screening methods for cervical cancer, both of them have limitations. This study aimed to develop and validate new methylation-based biomarkers from formalin-fixed paraffin-embedded (FFPE) tissue and TCT samples ina Chinese population to improve early detection of cervical cancer (CC) . Methods We performed a differential methylation analysis of cervical FFPE tissues and TCT samples through targeted methylation sequencing. Quantitative methylation-specific PCR (qMSP) was performed on cervical tissues, TCT samples and cell lines to construct a diagnostic model containing three highly methylated sites, namely JAM3, VSTM2B and EPHA7. The clinical effectiveness was evaluated on overall 279 TCT multiple group samples, comprising 33 squamous cell carcinomas (SCC), 14 adenocarcinoma(ADC), 43 high-grade squamous intraepithelial lesion (HSIL), 49 low-grade squamous intraepithelial lesion (LSIL), 78 high risk-HPV (HR-HPV)–positive/cytology-normal (NILM) cases, 32 healthy controls (NC), and 30 benign gynecological samples. The model performance was assessed using sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) . Results In an independent validation cohort (n = 108), the panel of three methylation-based biomarkers showed outstanding diagnostic accuracy. It achieved sensitivities of 100. 0% for SCC (n = 10), 80 . 0% for ADC (n = 5), and 86. 7% †Zhihan Liu, Guorong Liu and Shuanping Liu are contributed equally to this work. *Correspondence: Yongjun Li[li.yongjun@biochainbj.com](li.yongjun@biochainbj.com)[ ](li.yongjun@biochainbj.com)Xiaoliang Han [sean.han@biochainbj.com](sean.han@biochainbj.com)[ ](sean.han@biochainbj.com)Yang Cheng [eychengy@scut.edu.cn](eychengy@scut.edu.cn)[ ](eychengy@scut.edu.cn)Jing Yang [eyjingyang@scut.edu.cn](eyjingyang@scut.edu.cn) Full list of author information is available at the end of the article © The Author(s) 2026. Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit [http://creati](http://creati)[vecommons.org/licenses/by-nc-nd/4.0/](vecommons.org/licenses/by-nc-nd/4.0/.)[.](vecommons.org/licenses/by-nc-nd/4.0/.) Liu et al. Clinical Epigenetics (2026) 18:156 Page 2 of 11 for HSIL (n = 15) . Specificity was 90. 6% in HR-HPV–positive women (n = 32), 100. 0% in NC (n = 12), and 93. 3% in other beni

FAQs

What is the study trying to improve in cervical cancer screening?

The study aims to develop and validate methylation-based biomarkers to improve early detection of cervical cancer, addressing limitations of HPV testing and Thin-Prep Cytology Test (TCT).

How were the methylation biomarkers identified and measured?

Differential methylation analysis of cervical FFPE tissues and TCT samples was performed using targeted methylation sequencing, and quantitative methylation-specific PCR (qMSP) was used for measurement.

Which methylation sites are included in the diagnostic model?

The diagnostic model contains three highly methylated sites: JAM3, VSTM2B, and EPHA7 (combined as JAM3m/VSTM2Bm/EPHA7m).

How does the model perform in detecting cervical cancer and precancerous lesions?

In an independent validation cohort (n = 108), the three-biomarker panel shows outstanding diagnostic accuracy, with overall sensitivity of 90.0% and specificity of 92.3% reported in the validation results.

Sep 22, 2026

Upload by Clementine