Hepatocellular carcinoma (HCC) is the most common primary liver cancer, often diagnosed at advanced stages due to insufficient early screening and monitoring. MicroRNAs (miRNAs) are key regulators of gene expression and potential biomarkers for cancer diagnosis. This study investigated the diagnostic potential of miRNAs in Extracellular Vesicles (EVs) from HCC. miRNA expression in EVs was analyzed using HCC cell lines, circulating EVs from a Diethylnitrosamine (DEN)-induced liver tumor rat model, and plasma samples from HCC patients. Receiver Operating Characteristics (ROCs) were applied to evaluate the diagnostic accuracy of circulating EV miRNAs in patients. Five miRNAs (miR-183-5p, miR-19a-3p, miR-148b-3p, miR-34a-5p, and miR-215-5p) were consistently up-regulated in EVs across in vitro and in vivo HCC models. These miRNAs showed statistically significant differences in HCC patients stratified by TNM staging and Edmondson–Steiner grading compared to healthy controls. They also differentiated HCC patients with various etiologies from the control group and distinguished HCC patients, with or without liver cirrhosis, from cirrhotic and healthy individuals. Individually and as a panel, they demonstrated high sensitivity, specificity, and accuracy in identifying HCC patients. Their consistent upregulation across models and clinical samples highlights their robustness as biomarkers for HCC diagnosis, offering the potential for early disease management and prognosis.

Pleural mesothelioma usually occurs many years after asbestos fiber exposure; yet the mechanisms that convert chronic damage into malignancy remain unclear. Asbestos fibers induce persistent oxidative and genomic stress that should activate apoptosis via mitochondrial outer membrane permeabilization (MOMP). MOMP normally triggers cytochrome c (cyt c) release as well as mitochondrially derived damage-associated molecular patterns, resulting in downstream caspase activation, which leads to DNA damage and cell death. With sublethal activation, a phenomenon known as an “incomplete or minority MOMP (mMOMP)” occurs in which the cell survives the damage, enabling retention and propagation of somatic mutations. We tested whether prolonged asbestos fiber exposure drives mMOMP in the mesothelial cells, MeT-5A. After culturing the cells for 6 months with chrysotile asbestos fibers or crocidolite asbestos fibers, we observed an increase in clonogenicity, migration, and invasion, suggesting that transformation to a malignant-like phenotype was occurring. Next, we found increases in reactive oxygen species (ROS) production, cyt c release, and γH2AX phosphorylation without activation of caspase-3 and apoptotic induction. These cellular features are consistent with mMOMP. Additionally, an anti-apoptotic, mitochondrial protein, myeloid cell leukemia-1 was upregulated by asbestos and enabled mMOMP by preventing MOMP. Furthermore, we observed that asbestos-induced mMOMP was associated with features of drug-tolerant persister cells. mMOMP facilitates avoidance of apoptosis by mesothelial cells while acquiring malignant traits. Our study indicates that mMOMP is a mechanism that promotes carcinogenesis, mitochondrial changes, and metabolic reprogramming. These findings offer a foundation for future therapeutic investigations.

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.