The aim of this study is to evaluate the association between genes related to cellular senescence and ferroptosis and their relevance to hepatocellular carcinoma (HCC). Genes associated with senescence and ferroptosis in HCC were retrieved from public databases. A protein-protein interaction (PPI) network was constructed to identify for hub genes, and validate their expression. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis, while prognostic significance was determined through Kaplan-Meier analysis. A prognostic nomogram model was developed based on selected hub genes and Tumor Node Metastasis (TNM) staging. A total of 52 senescence-ferroptosis-related genes were identified in HCC. ROC analysis indicated moderate to high diagnostic efficacy for TP53 (AUC = 0.723, CI: 0.669-0.776), JUN (AUC = 0.733, CI: 0.659-0.806), RELA (AUC = 0.854, CI: 0.808-0.901), and CDKN2A (AUC = 0.953, CI: 0.932-0.974). Kaplan-Meier analysis revealed that TP53, HIF1A, and CDKN2A were significantly associated with overall survival (OS) in patients with HCC. A nomogram model incorporating these three genes and TNM staging achieved a concordance index (C-index) of 0.699. Calibration curves indicated concordance between the predicted and observed survival probabilities at 1-, 2-, and 3-year intervals. The senescence-ferroptosis-related genes TP53, HIF1A, and CDKN2A demonstrated potential as diagnostic and prognostic biomarkers in HCC. The developed nomogram may support individualized prognostic assessment and inform early diagnostic and therapeutic strategies in patients with HCC.

Merremia tridentata of the Convolvulaceae family exhibits multiple therapeutic properties and is commonly utilized in ethnomedicine. The anti-cancerous property of M. tridentata ethyl acetate extract (MtEAE) was evaluated in HepG2 hepatocellular carcinoma cells. Cytotoxicity was assessed using the MTT assay in a dose-dependent manner with an IC50 value of 57.55µg/mL. The PI and AO/EB staining revealed membrane damage and stimulation of cell death in treated cells. DNA damage was visualized by DAPI staining in treated cells. The Rhodamine 123 detects the mitochondrial membrane damage of the cells treated with MtEAE. The intracellular ROS distribution was assessed with DCFH-DA staining. The cell migration assay indicated suppression of cell migratory ability. Gene expression analysis showed augmented expression of p53 and BAX, a pro-apoptotic gene, along with downregulation of BCL-2, an anti-apoptotic gene indicating the induction of apoptosis. The hepatocellular carcinoma biomarker PCNA was downregulated causing inhibition of cell proliferation. Cell cycle analysis showed 71.82% of cells in the G0/G1 phase and it decreases further, confirming cell cycle arrest. These findings demonstrate that M. tridentata extract exerts potent anticancer activity by inducing apoptosis and inhibiting proliferation in HepG2 cells.

Plasma GPC3 reflects tumor GPC3 expression and predicts clinical outcomes in advanced HCC treated with atezolizumab + bevacizumab.

Development of a europium-salicylic acid based nano-biosensor for early detection of GPC3 in DEN-induced hepatocellular carcinoma.

High-performance electrochemical aptasensor for detection of glypican-3 based on nitrogen-doped reduced graphene oxide-ferrocene-polyaniline nanocomposites.

Hepatocellular carcinoma (HCC) could escape immune surveillance. Alpha-fetoprotein (AFP) serves as a significant biomarker for HCC; however, its influence on HCC immune surveillance remains elusive. RNA-Seq data of HCC were obtained from TCGA and GEO databases for the expression of AFP, MICA/B, and related genes. Immunohistochemistry for protein detection in tissues; the expression of target proteins was detected by Western blotting; membrane protein expression and cytotoxicity assessment were analysed by flow cytometry; protein localization was observed by immunofluorescence, cytokine levels were detected by ELISA; mRNA quantification was analysed by qRT-PCR, cell proliferation was measured by CCK-8, animal experiments were applied to observe immune response, and cytotoxicity assays were used to evaluate the killing effect of natural killer-92 (NK-92) cells. Results indicated that in both databases and patient tissues, AFP and MICA/B were highly expressed in the HCC tissues. AFP inhibits the membrane level of MICA/B in HCC cells and promotes the shedding of MICA/B by upregulating MMP9 expression via activation of the PI3K/AKT signalling pathway. Furthermore, AFP suppressed NK-92 cells from attacking HCC cells and restricted the release of cytokines by NK-92 cells, whereas interference with AFP had opposite effects. This finding indicated that AFP stimulated the cleavage of membrane MICA/B in HCC cells, increased the content of soluble MICA/B, and blocked the interaction between MICA/B and NKG2D, which may be involved in the upregulation of MMP9 expression via activation of the PI3K/AKT signalling pathway. These effects inhibited the activation of NK-92 cells, causing HCC cells to escape attack by NK-92 cells.

To evaluate the diagnostic performance of the Liver Imaging Reporting and Data System (LI-RADS) v2018 ancillary features (AFs) and biomarker for hepatocellular carcinomas (HCCs) ≤ 30 mm in Western and Eastern guidelines. This multicenter retrospective study included 1414 patients at risk for HCC who underwent MRI. The odds ratios of LI-RADS features and high alpha-fetoprotein (AFP) levels were assessed. Western and Eastern guidelines' diagnostic performances were compared via generalized estimating equations. A total of 1711 observations ≤ 30 mm were included in the extracellular contrast agent (ECA)-MRI training, internal/external ECA-MRI validation sets (n = 573, 245 and 212), internal EOB-MRI validation set (n = 681). Based on multivariable analysis, using combination of mild-moderate T2 hyperintensity and "either fat in mass or AFP ≥ 200 ng/mL," named "T2 + F/A" criterion, significantly improved the sensitivities of LR-5 v2018 (80.1-85.0% vs. 73.1-77.1%), EASL v2018 (78.4-84.3% vs. 72.2-75.7%), Korean Liver Cancer Association-National Cancer Center (KLCA-NCC) v2022 (78.4-90.9% vs. 73.4-88.3%), Asian Pacific Association for the Study of the Liver (APASL) v2017 (81.5-94.8% vs. 73.7-90.9%) and Japan Society of Hepatology (JSH) v2021 (81.5-94.5% vs. 73.7-90.6%) for HCCs ≤ 30 mm in the training set and internal and external validation sets (all p < 0.05) without impairing specificity, except for that of Asian Pacific Association for the Study of the Liver (APASL) v2017 in the internal gadoxetic acid (EOB) validation set (75.5% vs. 77.2%, p = 0.044). Two LI-RADS AFs (mild-moderate T2 hyperintensity and fat in mass) and AFP ≥ 200 ng/mL are useful for improving the sensitivity of Western and Eastern guidelines for HCCs ≤ 30 mm. Question The sensitivities of Western and Eastern guidelines for hepatocellular carcinoma ≤ 30 mm are still insufficient on extracellular contrast agent-enhanced MRI and gadoxetic acid-enhanced MRI. Findings The use of the integrated "T2 + F/A" diagnostic criterion significantly improved the sensitivity of Western and Eastern guidelines for HCCs ≤ 30 mm on both ECA-MRI and EOB-MRI. Clinical relevance Addressing a critical gap in noninvasive small HCC diagnosis protocols in Western and Eastern guidelines across multiple MRI contrast agents. By enabling more accurate MRI diagnosis of HCCs ≤ 30 mm, our findings could increase patients receiving curative treatments at the early stage, potentially improving outcomes.

FKBP10 as a prognostic biomarker and therapeutic target in hepatocellular carcinoma.

Endotrophin (ETP) is a cleavage fragment of collagen VI α3 (COL6A3) that functions as a potent fibrotic and pro-tumorigenic factor. ETP is a diagnostic and prognostic biomarker in hepatocellular carcinoma (HCC), continuously increasing throughout tumor development and promoting HCC progression. Elucidation of the underlying molecular mechanisms by which ETP exerts pro-tumorigenic effects in the liver could uncover potential therapeutic strategies. Using peroxidase-catalyzed proximity labeling, we identified CD44 as an ETP receptor. ETP binding to CD44 activated STAT3 signaling, promoting epithelial-mesenchymal transition (EMT), proliferation, and sorafenib resistance. Hepatic stellate cell-derived ETP targeted pericentral CD44+ tumor cells, inducing COL6A3 expression and sustaining ETP production via a STAT3-dependent feedback loop. Disruption of this axis by CD44 knockout, STAT3 inhibition, or CD44-binding-deficient ETP mutants suppressed malignant phenotypes in vitro. In metabolic dysfunction-associated HCC induced by diethylnitrosamine (DEN) plus high-fat diet (HFD), dual knockout of Col6a3 and Cd44 in mice markedly reduced tumor burden, restored sorafenib sensitivity, and attenuated EMT, fibrosis, and steatotic-fibrotic niche formation. These findings establish the ETP-CD44-STAT3 axis as a driver of tumor-stroma crosstalk linking fibro-inflammation to malignancy, highlighting it as a therapeutic target in obesity-associated liver cancer.

Hepatobiliary phase (HBP) findings on Gd-EOB-DTPA-enhanced MRI (EOB-MRI) have been proposed as biomarkers of hepatocellular carcinoma (HCC). However, the association between intratumor signal heterogeneity in HBP of EOB-MRI and survival or tumor microenvironment remains unclear. We retrospectively analyzed 167 patients who underwent hepatic resection for HCC. The coefficient of variation (CV) in HBP on preoperative EOB-MRI was calculated as the standard deviation/mean of the intratumor signal intensity. Hyperintense area (HIA) was defined as tumor regions with signal intensity equal to or higher than the surrounding parenchyma. Associations of imaging-defined heterogeneity (high CV with HIA) with survival and immune infiltration were evaluated. Multivariate analyses showed male, negative HBs-antigen, elevated PIVKA-II, multiple tumors, microvascular invasion, and high CV with HIA (all p < 0.05) were significant predictors of disease-free survival, while negative HBs-antigen, elevated PIVKA-II, microvascular invasion, and high CV with HIA (all p < 0.05) were significant predictors of overall survival. High CV with HIA correlated with male (p = 0.04), larger tumor size (p < 0.01), multiple tumors (p < 0.01), and less CD8+ T-cell infiltration into tumor center (p = 0.03) and invasive margin (p = 0.048). High CV with HIA in HBP of EOB-MRI may serve as a novel prognostic imaging biomarker after resection and reflect an immune-cold HCC microenvironment.

Alpha-fetoprotein acts as a key regulator of cancer stemness in hepatocellular carcinoma via PI3K/Akt pathway.

Hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide, and early diagnosis is crucial. Golgi protein 73 (GP73) has emerged as a promising serum biomarker for HCC. However, current detection methods often fail to meet routine screening requirements due to limitations in sensitivity and operational simplicity. To address these challenges, we have developed a novel fluorescent aptamer-based sensor for highly sensitive GP73 detection based on the fluorescence resonance energy transfer (FRET) mechanism between graphitic carbon nitride quantum dots (g-CNQDs) and a copper-based metal-organic framework (Cu-TCPP). g-CNQDs were covalently conjugated with a GP73-specific aptamer to serve as the fluorescent donor, while two-dimensional Cu-TCPP nanosheets acted as the efficient acceptors. Fluorescence was quenched upon donor-acceptor interaction via FRET. In the presence of GP73, aptamer-target binding disrupted FRET interaction, separating the donor from the acceptor and restoring fluorescence in a concentration-dependent manner. Under optimal conditions, the sensor exhibited excellent linearity over a concentration range 1.0-225.0 ng mL⁻¹, with a detection limit as low as 0.907 ng mL⁻¹. Recoveries for spiked human serum samples ranged from 95.96% to 103.85%, with relative standard deviations (RSDs) of 0.38%-5.48%. The developed aptamer sensor demonstrated excellent sensitivity, selectivity, and stability, providing a powerful tool for early HCC diagnosis and offering strong potential for real-time analytical applications.

BACKGROUND Recent research has highlighted DNA methylation as a promising diagnostic biomarker for hepatocellular carcinoma (HCC). Fatty Acyl-CoA Reductase 1 (FAR1) exhibits a high propensity for methylation in HCC. This study aimed to evaluate diagnostic and prognostic potential of FAR1 methylation in liver transplantation (LT) recipients with HCC. MATERIAL AND METHODS This analysis used droplet digital polymerase chain reaction to quantify FAR1 methylation levels in stored pretransplant blood samples. The study cohort (n=48) comprised 25 liver cirrhosis patients with HCC, 13 with cirrhosis but no HCC, and 10 healthy donors. RESULTS Median and mean methylation levels of FAR1 in these groups were 4 copies, zero copies, and zero copies, and 31.6±74.5, 1.5±3.5, and 0.1±0.4 copies, respectively (p<0.001). Receiver operating characteristic curve analysis revealed area under the curve of 0.832 for FAR1, outperforming a-fetoprotein (AFP; 0.737) and protein induced by vitamin K absence or antagonist-II (PIVKA-II; 0.732). A cut-off value of 1 copy for FAR1, defined by Youden's Index (J=0.599), yielded sensitivity of 82.6% and specificity of 77.3%, surpassing diagnostic capacities of AFP and PIVKA-II. Combining FAR1 >1 copy with AFP >7.5 ng/mL or PIVKA-II >40 mAU/mL increased the sensitivity to 91.3%, with specificity of 72.7% and overall accuracy of 82.2%. There was no significant correlation between FAR1 methylation levels and tumor recurrence or overall survival when using a cut-off of 1 copy. CONCLUSIONS These findings suggest that FAR1 methylation is a valuable biomarker for diagnosing HCC in patients with advanced liver disease awaiting transplantation. Further large-scale investigations are necessary to validate clinical efficacy.

Integrated and clinical validation of helicase-like transcription factor as a biomarker for hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) and chronic liver disease (CLD) are mostly linked to hepatitis C virus (HCV) infection, especially genotype 4. The advancement of liver cancer is significantly influenced by epigenetic mechanisms, such as DNA methylation and microRNA regulation. This study aimed to assess the expression of two tumor suppressor genes in patients infected with HCV genotype 4: glutathione S-transferase pi 1 (GSTP1) and E-cadherin 1 (CDH1). It also sought to investigate the regulatory interactions among these genes, microRNA152 (miRNA152), and DNA methyltransferase 1 (DNMT1) to assess their potential as biomarkers for hepatocellular carcinoma (HCC) development. Quantitative real-time PCR was done to evaluate the gene expression levels of GSTP1, CDH1, miRNA152, and DNMT1 in CLD and HCC patients infected with HCV genotype 4. Correlation analyses were used to examine regulatory linkage. The results indicated a substantial inverse correlation between DNMT1 and miRNA152. Furthermore, DNMT1 expression exhibited a negative association with both GSTP1 and CDH1, while GSTP1 and CDH1 had a positive correlation. The results indicate that DNMT1 and miRNA152 govern the epigenetic inactivation of tumor suppressor genes. In individuals with HCV genotype 4, the interplay of miRNA152, DNMT1, GSTP1, and CDH1 may contribute to the pathogenesis of HCC. These indicators demonstrate potential roles as therapeutic targets and noninvasive prognostic biomarkers for HCV-related liver disease.

Emerging evidence highlights the significant role of Neutrophil Extracellular Traps (NETs) in hepatocellular carcinoma (HCC), but the underlying molecular mechanisms involving NETs formation remain incompletely understood. This study aims to identify key biomarkers related to NETs in HCC through bioinformatic analysis. We obtained RNA sequencing data of hepatocellular carcinoma tissues and adjacent normal liver tissues from the Gene Expression Omnibus (GEO) databases, followed by data correction, integration, and annotation. Subsequently, differentially expressed genes (DEGs) were identified, and Weighted Gene Co-expression Network Analysis (WGCNA) was used to screen for disease-related genes. The intersection with NETs-related genes (NRGs) yielded differentially expressed NET-related genes (DENRGs), which were subjected to single-sample Gene Set Enrichment Analysis (ssGSEA). Three machine learning models (LASSO, SVM-RFE, and RF) were further employed to screen for key biomarkers. Receiver Operating Characteristic (ROC) curves and a nomogram model were used to validate the diagnostic and predictive efficacy of those key biomarkers, with further validation using an external dataset. Unsupervised clustering and Gene Set Variation Analysis (GSVA) were performed on the key biomarkers. We conducted a comprehensive bioinformatic analysis on 223 HCC samples and 127 normal liver tissue samples from 5 datasets. Transcriptomic analysis identified 826 DEGs. WGCNA revealed key gene modules associated with HCC, including 362 genes. By intersecting these with 627 NRGs, we identified 18 DENRGs. The results of ssGSEA showed that most of immune cells were significantly downregulated in HCC. Machine learning models (LASSO, SVM-RFE, and RF) identified three downregulated biomarkers (ECM1, DNASE1L3, JUN). A nomogram and ROC curves confirmed the diagnostic accuracy of these biomarkers. Cluster analysis revealed two distinct HCC subtypes with different immune microenvironment characteristics. Drug-gene interaction analysis identified potential inhibitors targeting DNASE1L3 and JUN. This study identified NET-related key biomarkers (ECM1, DNASE1L3, JUN) as reliable diagnostic tools for HCC, highlighting their diagnostic and therapeutic potential, and providing insights for HCC diagnostic tools and immunotherapy strategies.

Circulating tumor cells (CTCs) are emerging as promising biomarkers in hepatocellular carcinoma (HCC), offering noninvasive insight into tumor biology, progression, and treatment response. Advances in enrichment and single-cell technologies have enabled the characterization of CTCs at genomic, transcriptomic, epigenetic, and proteomic levels. CTC detection correlates with tumor stage, vascular invasion, and alpha-fetoprotein levels, supporting their role in early diagnosis and staging. Specific surface markers such as epithelial cell adhesion molecule, GPC3, ASGPR, and stem cell-associated antigens like CD90 and CD133 help classify CTC subtypes with prognostic and therapeutic relevance. Mesenchymal and hybrid phenotypes, identified via epithelial-to-mesenchymal transition markers, are linked to recurrence and metastasis. Furthermore, CTCs provide a platform for companion diagnostics by reflecting mutational and resistance profiles, particularly in the context of targeted therapies and immune checkpoint inhibitors. Despite current limitations in standardization, sensitivity, and scalability, the integration of CTC analysis into clinical practice holds potential to enhance precision medicine strategies in HCC.

Pathological features and biomarkers of hepatocellular carcinoma: a bibliometric analysis from 2005 to 2025

Multi-omics analysis of BTF3L4 as a prognostic and immune biomarker in hepatocellular carcinoma

Matrix stiffness-induced NARF promotes hepatocellular carcinoma progression by enhancing LEF1-mediated transcription.