miRNAs-set of Plasmatic Extracellular Vesicles as Novel Biomarkers for Hepatocellular Carcinoma Diagnosis

Hepatocellular carcinoma (HCC) is the most common liver cancer, often diagnosed at advanced stages due to inadequate early diagnostic methods. MicroRNAs (miRNAs) play crucial roles in gene expression regulation and are possible biomarkers for molecular diagnosis in several diseases, including cancer. This study aimed to investigate the diagnostic potential of miRNAs contained in Extracellular Vesicles (EVs) from HCC. The miRNA expression contained in EVs was analyzed in HCC cell lines, circulating EVs in a Diethylnitrosamine (DEN)-induced HCC rat model, and plasma samples of HCC patients. Receiver Operating Characteristics (ROC) were applied to circulating EV-miRNAs from human patients to evaluate their diagnostic accuracy. The miRNA expression analysis identified five miRNAs (miR-183-5p, miR-19a-3p, miR-148b-3p, miR-34a-5p, and miR-215-5p) consistently up-regulated in EVs from in vitro and in vivo HCC models. These five EVs-derived miRNAs showed statistically significant differences in HCC patients stratified by TNM staging and Edmondson-Steiner grading compared to healthy controls. Individually and in combination, they demonstrated high sensitivity, specificity, and accuracy in distinguishing HCC patients from healthy subjects. The consistent upregulation of the five miRNAs across different experimental models and clinical samples suggests their robustness as biomarkers for HCC diagnosis, and it underscores their clinical potential in early disease management and prognosis.