Xu Hang, Wang Min, Wu Shuting, Li Qinke, Wang Jinlong, Zhang Siying, Liu Qiongming, Wang Mengting, Li Ruifang, Hu Zhiyuan, Liu Yi, Yang Zhu
Department of Gynecology and Obstetrics, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Fujian Provincial Key Laboratory of Brain Aging and Neurodegenerative Diseases, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian, China.
Front Oncol. 2025 Jul 28;15:1563095. doi: 10.3389/fonc.2025.1563095. eCollection 2025.
Epithelial ovarian cancer (EOC) currently lacks highly specific biomarkers for clinical screening. This study aimed to identify and validate novel auxiliary diagnostic markers for EOC.
Through integrated analysis of transcriptome sequencing data and single-cell RNA sequencing from public databases, we identified mesothelin (MSLN) as an EOC-specific target. MSLN expression was subsequently validated in EOC cell lines and clinical specimens by flow cytometry, immunofluorescence, and immunohistochemistry. The capture efficacy of Pep@MNPs (Magnetic nanoparticles functionalised with EpCAM peptides) on EOC cells was verified by scanning electron microscopy, Prussian blue staining and cell spiked-blood capture experiments. In a prospective cohort of 35 patients with undiagnosed ovarian masses, we employed immunofluorescence staining to detect MSLN-positive circulating tumor cells (MSLN(+)CTCs) and assessed their diagnostic performance using receiver operating characteristic (ROC) analysis.
MSLN was highly expressed in EOC cell line and tissues but lowly expressed in normal ovarian surface epithelial tissues. EOC cells can be captured by Pep@MNPs with high sensitivity and specificity. ROC curves analysis showed that MSLN(+)CTCs differentiated between benign and malignant lesions of the ovary with a sensitivity of 66.67% and a specificity of 95% (p = 0.0014), which was more specific than cancer antigen 125 (CA125) (sensitivity: 71.43%; specificity: 94.47%; p < 0.0001) and human epididymis protein 4 (HE4) (sensitivity: 84.62%; specificity: 89.47%; p = 0.0002). When MSLN(+)CTCs were combined with CA125, the sensitivity was 92.86% and the specificity was 94.74%, p < 0.0001, which greatly improved the diagnostic sensitivity while preserving high specificity.
MSLN(+)CTCs represent a highly specific auxiliary biomarker for differentiating benign and malignant ovarian lesions. The combination of MSLN(+)CTCs with CA125 provides an optimal balance between sensitivity and specificity, offering promising clinical utility for EOC diagnosis.
上皮性卵巢癌（EOC）目前缺乏用于临床筛查的高度特异性生物标志物。本研究旨在鉴定和验证EOC的新型辅助诊断标志物。
通过对公共数据库中的转录组测序数据和单细胞RNA测序进行综合分析，我们将间皮素（MSLN）鉴定为EOC特异性靶点。随后通过流式细胞术、免疫荧光和免疫组织化学在EOC细胞系和临床标本中验证MSLN表达。通过扫描电子显微镜、普鲁士蓝染色和细胞加样血液捕获实验验证了Pep@MNPs（用EpCAM肽功能化的磁性纳米颗粒）对EOC细胞的捕获效率。在一个由35例未确诊卵巢肿块患者组成的前瞻性队列中，我们采用免疫荧光染色检测MSLN阳性循环肿瘤细胞（MSLN(+)CTCs），并使用受试者工作特征（ROC）分析评估其诊断性能。
MSLN在EOC细胞系和组织中高表达，但在正常卵巢表面上皮组织中低表达。Pep@MNPs能以高灵敏度和特异性捕获EOC细胞。ROC曲线分析显示，MSLN(+)CTCs区分卵巢良性和恶性病变的灵敏度为66.67%，特异性为95%（p = 0.0014），比癌抗原125（CA125）（灵敏度：71.43%；特异性：94.47%；p < 0.0001）和人附睾蛋白4（HE4）（灵敏度：84.62%；特异性：89.47%；p = 0.0002）更具特异性。当MSLN(+)CTCs与CA125联合使用时，灵敏度为92.86%，特异性为94.74%，p < 0.0001，在保持高特异性的同时大大提高了诊断灵敏度。
MSLN(+)CTCs是区分卵巢良性和恶性病变的高度特异性辅助生物标志物。MSLN(+)CTCs与CA125的联合使用在灵敏度和特异性之间提供了最佳平衡，为EOC诊断提供了有前景的临床应用价值。
