Cervical cancer (CC) remains the fourth most common cancer and the fourth leading cause of cancer death in women worldwide. While tests for high-risk human papillomavirus (HR-HPV) have improved the sensitivity of cervical cancer screening, their limited specificity makes it difficult to distinguish between transient infections and those likely to progress to high-grade cervical intraepithelial neoplasia (CIN), the main precursor to CC. Therefore, additional biomarkers are needed to improve risk stratification in women with HR-HPV. Dysregulated microRNA (miRNA) expression plays a crucial role in HR-HPV-induced cervical carcinogenesis, representing a promising source of molecular biomarkers.
Objective: To identify a miRNA expression signature with potential utility as a non-invasive biomarker for triaging HR-HPV-positive women and improving the detection of high-grade cervical lesions.
Methods: In this cross-sectional study, cervical scraping samples from women with high-risk HPV were histologically classified as low-grade (CIN1) or high-grade (CIN2/3) lesions. Total RNA was extracted, and small RNA sequencing was performed using the Illumina MiSeq platform. Differentially expressed miRNAs were identified and analyzed using unsupervised clustering, ROC curve analysis, logistic regression, and in silico functional enrichment. Selected miRNAs were validated by reverse transcription quantitative polymerase chain reaction (RT-qPCR).
Results: Nineteen miRNAs with differential expression between CIN1 and CIN2/3 lesions were identified. Four miRNAs (hsa-miR-1271-5p, hsa-miR-9-5p, hsa-miR-501-3p, and hsa-miR-1246) were associated with viral and oncological pathways and were confirmed during the validation phase. Among them, hsa-miR-501-3p demonstrated high diagnostic performance for the detection of CIN2/3 (AUC = 0.88), which was further improved when combined with hsa-miR-1271-5p and hsa-miR-9-5p (AUC = 0.94).
Conclusion: This study identifies a miRNA expression signature with high discriminatory power for high-grade cervical lesions in women with high-risk HPV. These findings support the potential clinical integration of miRNA-based biomarkers to improve triage strategies in cervical cancer screening programs