Engrailed‑2 (EN2) protein in cervical mucus - a novel biomarker for endometrial carcinoma

Q: What was the main objective of the study?

To determine whether EN2 protein in cervical mucus can function as a novel biomarker for screening endometrial cancer.

Q: How were EN2 protein levels measured and validated?

Cervical mucus EN2 levels were measured by ELISA in patient groups, and expression differences were further verified using RT-PCR, immunohistochemistry, and western blotting, including cell-line analyses.

Q: What diagnostic performance did the study report?

ROC analysis identified an optimal threshold of 321.1 ng/mL for cervical mucus EN2, with sensitivity of 92.6% and specificity of 95.4% to distinguish EINIII, EC, and non-cancerous cases.

Q: What does the study conclude about clinical use?

Higher EN2 concentration in cervical mucus can effectively identify endometrial cancer and precancerous lesions, though larger multi-center studies are needed to confirm the diagnostic role.

Sep 21, 2026

Uploaded by Ophelia

A study evaluated whether EN2 protein levels in cervical mucus can serve as a novel noninvasive biomarker for screening endometrial cancer. Samples were collected before hysteroscopy from 133 patients, grouped by endometrial pathology, and EN2 was quantified by ELISA, with additional validation using RT-PCR, immunohistochemistry, and western blotting, plus cell-line overexpression and expression analyses. EN2 was higher in endometrial cancer and precancerous lesions, and ROC-derived thresholds supported diagnostic discrimination.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages10

Reading Time25 min

AccessFree to download

Abstract

Introduction

Materials and Methods

Results

Conclusion

Clinical and Translational Oncology (2025) 27:2484–2493 [https://doi.org/10.1007/s12094-024-03799-5](https://doi.org/10.1007/s12094-024-03799-5) Engrailed‑2 (EN2) protein in cervical mucus: a novel biomarker for endometrial carcinoma Tong Wang1 · Ningyi Jia1 · Songkun Gao1 · Shengjie Liu2,4 · Ming Liu2,4 · Hong Zhang3 Received: 7 September 2024 / Accepted: 13 November 2024 / Published online: 28 November 2024 © The Author(s) 2024 Abstract Objective This study aims to demonstrate that the EN2 protein in cervical mucus may serve as a novel biomarker for screening endometrial cancer. Materials and Methods This study included 133 patients who were treated at Beijing Obstetrics and Gynecology Hospital. According to the pathological results of hysteroscopy endometrial biopsy, the patients were divided into endometrial cancer group (n = 55), endometrial atypical hyperplasia group (n = 16), benign lesion group (n = 28), and control group (n = 34) . All patients collected cervical mucus before hysteroscopy, and the level of EN2 protein in cervical mucus was detected by ELISA in the four groups of patients. Nine patients who underwent surgical treatment for endometrial cancer were included, and endometrial cancer lesions and adjacent tissues without endometrial cancer infiltration were taken from the endometrial cancer lesions and cervical tissues. Fifteen patients who underwent hysterectomy for benign lesions were included, and endometrial and cervical tissues were taken from the endometrial cancer lesions and adjacent tissues without endometrial cancer infiltration. PT-PCR, immunohistochemistry, and WB were used to verify the expression differences of EN2 protein in cancerous lesions and endometrial tissues without endometrial infiltration, cervical tissues, and endometrial tissues without endometrial lesions. In addition, HEC1B, KLE, and HeLa cell lines were used to characterize EN2 overexpression in endometrial cancer cells. Immunohistochemistry, RT-PCR, and confocal analysis were used to detect the expression of EN2 protein in different cell lines. Results In different groups, the average concentration of EN2 protein in cervical mucus in the EC group was significantly higher than that in the benign group and the normal control group (573.9 ± 123.4 ng/mL vs 153.5 ± 106.2 ng/mL and 153.0 ± 107.5 ng/mL, P < 0.001) . The concentration of EN2 protein in EIN3 case specimens was significantly higher than that in the normal control group (P < 0.001) . ROC analysis showed that the optimal threshold for diagnosing endometrial cancer in cervical mucus was 321.1 ng/ml, with a sensitivity of 92.6% and specificity of 95.4% for distinguishing EINIII, EC, and non-cancerous cases. In clinical specimens, the expression level of EN2 protein in endometrial cancer tissues was significantly higher than that in endometrial tissues and cervical tissues without endometrial cancer infiltration. In addition, PT-PCR, immunohistochemistry, suggest that EN2 protein is overexpressed in endometrial cancer cell lines compared to cervical adenocarcinoma cells (P < 0 .01) . Conclusion The concentration of EN2 protein in cervical mucus can effectively identify endometrial cancer and precancerous lesions. The increased expression of EN2 protein in endometrial cancer lesions leads to an increase in the concentration of EN2 protein in the cervical mucus of endometrial cancer patients.This is a small single-center study, and larger studies are needed to determine the role of EN2 protein in the diagnosis of endometrial cancer. Keywords Engrailed-2 · Cervical mucus · Endometrial carcinoma · Biomarker · Diagnosis Introduction Endometrial carcinoma (EC) is one of the most common gynecological cancers. 382 069 new cases of corpus uteri cancer and 89 929 women died from the disease allover the world in 2018 [1] . 5-year survival for early-stage disease is 69–90%, while 5-year survival decreases to 17–58% for stages III-IV. Early detection of endometri

FAQs

What was the main objective of the study?

To determine whether EN2 protein in cervical mucus can function as a novel biomarker for screening endometrial cancer.

How were EN2 protein levels measured and validated?

Cervical mucus EN2 levels were measured by ELISA in patient groups, and expression differences were further verified using RT-PCR, immunohistochemistry, and western blotting, including cell-line analyses.

What diagnostic performance did the study report?

ROC analysis identified an optimal threshold of 321.1 ng/mL for cervical mucus EN2, with sensitivity of 92.6% and specificity of 95.4% to distinguish EINIII, EC, and non-cancerous cases.

What does the study conclude about clinical use?

Higher EN2 concentration in cervical mucus can effectively identify endometrial cancer and precancerous lesions, though larger multi-center studies are needed to confirm the diagnostic role.

Sep 21, 2026

Uploaded by Ophelia