Combined plasma and liquid-based cytology microRNA signatures as biomarkers of high-grade cervical lesions: association with HPV infection and p16 expression

MicroRNA (miRNA) dysregulation may provide complementary molecular information for cervical lesion assessment and triage. This prospective observational study evaluated six candidate miRNAs in paired plasma and liquid-based cytology (LBC) specimens and their associations with cytological severity, high-risk human papillomavirus (hrHPV), and p16 expression. Among 328 women, cytology was classified as normal (n=52), atypical squamous cells of undetermined significance (ASC-US; n=44), low-grade squamous intraepithelial lesion (LSIL; n=42), high-grade squamous intraepithelial lesion (HSIL; n=82), or squamous cell carcinoma (SCC; n=108). hrHPV positivity increased from 0% in normal cytology to 93.5% in SCC and p16 positivity from 0% to 95.4% (both P<0.001). In paired plasma and LBC specimens, miR-21-5p, miR-155-5p, miR-9-5p, and miR-15b-5p progressively increased with cytological severity, whereas miR-375-3p and miR-145-5p decreased (all p<0.001). For discrimination of HSIL/SCC from normal/ASC-US/LSIL, individual-miRNA areas under the curve (AUCs) ranged from 0.970 to 0.993. After stratified 10-fold cross-validation, AUCs were 0.818 (95% CI, 0.765-0.869) for hrHPV, 0.807 (0.751-0.860) for p16, and 0.840 (0.789-0.888) for hrHPV+p16. The six-miRNA LBC panel achieved an AUC of 0.9999 (0.9996-1.000), 99.5% sensitivity, 100.0% specificity, and a Brier score of 0.006. These findings demonstrate strong internal discrimination by LBC-derived miRNAs; however, validation against histologically confirmed endpoints in independent, clinically representative populations is required before clinical application.