BACKGROUND:Gastric cancer (GC) remains one of the leading causes of cancer-related mortality worldwide. Ubiquitin-conjugating enzyme E2 K (UBE2K), defined as an E2, is involved in various cellular processes. N6-methyladenosine (m6A) is one of the most abundant subtypes of RNA modifications. However, the systematic role of UBE2K and whether UBE2K undergoes an m6A modification in GC remain unknown. METHODS:The expression and prognosis of UBE2K in GC were analyzed through an online database. RT-qPCR, western blot and IHC were used to test the expression of UBE2K in GC tissues and cells. CCK-8, colony formation, transwell, wound healing and sphere formation assays were conducted to explore UBE2K's function in vitro. The subcutaneous mouse model was generated to validate UBE2K's role in tumorigenesis. RNA immunoprecipitation and RNA stability experiment were applied to investigate the molecular mechanism of UBE2K. RESULTS:Higher expression of UBE2K was found in GC tissues and predicted worse prognosis. UBE2K knockdown suppressed the malignant progression of GC cells both in vitro and in vivo. Mechanistically, we found that UBE2K was regulated by m6A modification. WTAP was further revealed as the m6A writer of UBE2K to enhance UBE2K RNA stability. Finally, functional rescue experiments showed that silencing WTAP reversed the oncogenic effects of UBE2K overexpression on GC cells, whereas WTAP attenuated the suppression of GC cells induced by UBE2K knockdown. CONCLUSION:Our findings indicate WTAP-mediated m6A modification upregulates UBE2K and the WTAP/UBE2K axis facilitates GC progression, suggesting a potential therapeutic target for GC treatment.

BACKGROUND:LRRC41 is critical for the progression of multiple cancers, especially hepatocellular carcinoma (HCC), yet its oncogenic mechanism in HCC remains unclear. This study aimed to explore the molecular mechanism by which LRRC41 promotes HCC malignancy and investigate the therapeutic potential of inhibiting its mediated signalling pathway. METHODS:LRRC41 expression in HCC was analysed via the TCGA, HPA databases and experimental detection. Its biological functions were assessed by in vitro CCK-8, Transwell, colony formation assays and in vivo xenograft models. IP-MS, Co-IP and ubiquitination assays were used to elucidate the molecular mechanism, and the anti-tumour effect of the USP7 inhibitor P6620 was verified in vitro and in vivo. RESULTS:LRRC41 was abnormally overexpressed in HCC and correlated with poor prognosis, promoting HCC cell proliferation, invasion and tumorigenesis. USP7 stabilised LRRC41 via deubiquitination, and LRRC41 activated the NF-κB pathway by targeting HNRNPC in a K63-linked ubiquitination-dependent manner. P6620 inhibited the USP7/LRRC41 axis to suppress HCC malignancy, and its combination with lenvatinib enhanced in vivo anti-tumor efficacy. CONCLUSIONS:LRRC41 overexpression drives HCC progression and poor prognosis; the novel USP7/LRRC41/HNRNPC/NF-κB axis is identified in HCC. USP7 inhibitor P6620 blocks this axis and may serve as a potential agent for HCC combination chemotherapy. The regulatory mechanism of the USP7/LRRC41/HNRNPC/NF-κB signalling axis in HCC. Created with Figdraw.

Background Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy with a complex tumor ecosystem that contributes to its progression. Deubiquitinases (DUBs) are vital regulators in cancer. However, the overall activity of DUBs and their role in driving PDAC progression within immune microenvironment remain largely unknown.Methods We employed an integrative multi-omics strategy combining machine learning (ML) on bulk transcriptomic data, single-cell RNA sequencing and spatial transcriptomic profiling. We applied Coxnet and Fuzzy SVM for prognostic modeling, inferCNV for malignant cell identification, SCENIC for transcription factor regulon analysis, LIANA+ for inferring inter-cellular communication networks and cell2location for spatial deconvolution. USP54 expression was detected by real-time quantitative PCR, western blotting and immunohistochemistry. USP54 function was validated through in vitro and in vivo assays.Results ML-based pathway analysis revealed post-translational modification as a major prognostic category, within which elevated DUBs activity emerged as an independent adverse prognostic factor. At the single-cell level, USP54 was upregulated along the trajectory of malignant ductal cells and correlated with an inflamed tumor microenvironment. Cell-cell communication analysis predicted signaling from monocytes/macrophages to tumor cells via the THBS1-integrin ligand-receptor pair. This immune-derived signaling potentially converged on KLF5-positive tumor cells, with KLF5 identified as a putative transcriptional activator of USP54. Spatial transcriptomics validated the co-localization of USP54 expression, elevated DUB activity, and KRAS signaling within specific tumor niches adjacent to THBS1-enriched immune regions. High USP54 expression was frequently observed in PDAC tissues and associated with poor patient survival. More importantly, in both BxPC-3 and PANC-1 cell lines, USP54 knockdown suppressed cell proliferation and metastasis, whereas its overexpression enhanced these malignant phenotypes. Subcutaneous xenograft growth and tail vein injection experiments validated these findings in vivo.Conclusions Our comprehensive multi-omics analysis and experimental validation identify the deubiquitinase USP54 as a novel promoter of PDAC progression within a spatially organized tumor-immune microenvironment. These findings suggest USP54 as both a candidate prognostic biomarker and a potential therapeutic target for this lethal malignancy.

Background:Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide and its incidence and mortality rates remain high. Therefore, new diagnostic and therapeutic approaches are urgently required. Family with sequence similarity 188 member B (FAM188B) encodes an evolutionarily conserved protein that is highly expressed in various cancers. While FAM188B has been implicated in the progression of several tumors, its role in HCC progression remains unknown. Methods:We analyzed FAM188B expression in HCC using The Cancer Genome Atlas (TCGA) and The University of Alabama at Birmingham Cancer data analysis Portal (UALCAN) databases. Functional studies included in vitro proliferation, migration, and invasion assays, as well as in vivo xenograft models. Co-immunoprecipitation (Co-IP), Western blotting, and immunofluorescence were used to investigate the FAM188B-Ubiquitin-specific peptidase 10 (USP10)-Yes-associated protein/Transcriptional coactivator with PDZ-binding motif (YAP/TAZ) interaction. Results:FAM188B was found highly expressed in HCC cells and associated with poor prognosis. Both in vitro and in vivo, FAM188B promoted the proliferation, migration, and invasion of HCC. FAM188B directly interacts with and stabilizes USP10 and the downregulation of FAM188B by shRNA led to decreased USP10 and YAP/TAZ protein levels, suggesting that FAM188B may regulate the YAP/TAZ pathway through its interaction with USP10. Conclusion:Our findings reveal that FAM188B plays a crucial role in enhancing HCC cell proliferation, migration, and invasion, primarily through regulating the USP10/YAP/TAZ signaling axis, which was validated in vitro and in vivo.

Hepatocellular carcinoma (HCC) is a highly aggressive malignancy of the digestive system characterized by increasing global incidence and mortality rates. Translocase of the Inner Mitochondrial Membrane 23 (TIMM23), a key component of the mitochondrial inner membrane translocase complex, plays a critical role in the import and localization of mitochondrial proteins. Elevated TIMM23 expression is significantly associated with poor prognosis in patients with HCC Conversely, TIMM23 downregulation was found to have induced apoptosis and significantly inhibited the proliferation, migration, and invasive potential of HCC cells both in vitro and in vivo. Furthermore, we have identified the functional interplay between TIMM23 and ERA G-protein-like 1 (ERAL1). TIMM23 knockdown markedly altered mitochondrial membrane potential and permeability, leading to a subsequent decrease in ERAL1 expression. ERAL1 interacts with Bcl-2-like protein 1 (Bcl-XL) during apoptosis. These findings collectively underscore the pivotal role of the TIMM23-ERAL1 axis in HCC progression, suggesting that the therapeutic targeting of TIMM23 may offer a promising strategy for HCC treatment.

Background Hepatic ischemia-reperfusion injury (HIRI) is a pathophysiological process during liver transplantation, characterized by insufficient oxygen supply and subsequent restoration of blood flow leading to an overproduction of reactive oxygen species (ROS), which in turn activates the inflammatory response and leads to cellular damage. Therefore, reducing excess ROS production in the hepatic microenvironment would provide an effective way to mitigate oxidative stress injury and apoptosis during HIRI. Nanozymes with outstanding free radical scavenging activities have aroused great interest and enthusiasm in oxidative stress treatment. Results We previously demonstrated that carbon-dots (C-dots) nanozymes with SOD-like activity could serve as free radicals scavengers. Herein, we proposed that C-dots could protect the liver from ROS-mediated inflammatory responses and apoptosis in HIRI, thereby improving the therapeutic effect. We demonstrated that C-dots with anti-oxidative stress and anti-inflammatory properties improved the survival of L-02 cells under H 2 O 2 and LPS-treated conditions. In the animal model, Our results showed that the impregnation of C-dots could effectively scavenge ROS and reduce the expression of inflammatory cytokines, such as IL-1β, IL-6, IL-12, and TNF-α, resulting in a profound therapeutic effect in the HIRI. To reveal the potential therapeutic mechanism, transcriptome sequencing was performed and the relevant genes were validated, showing that the C-dots exert hepatoprotective effects by modulating the hepatic inflammatory network and inhibiting apoptosis. Conclusions With negligible systemic toxicity, our findings substantiate the potential of C-dots as a therapeutic approach for HIRI, thereby offering a promising intervention strategy for clinical implementation.

Hepatocellular carcinoma (HCC) is characterized by a profoundly hypoxic microenvironment, which drives tumor aggressiveness and poor clinical outcomes. However, the precise regulatory mechanisms through which hypoxia promotes HCC progression remain incompletely understood. Here, we identified ubiquitin conjugating enzyme E2 variant 1 (UBE2V1) as a novel hypoxia-responsive gene that is transcriptionally activated by hypoxia-inducible factor-1α (HIF-1α) through direct binding to a hypoxia-response element located between −208 and −201 bp in the UBE2V1 promoter. Overexpression of UBE2V1 was frequently detected in HCC tissues and correlated strongly with advanced tumor stage and unfavorable patient prognosis. Moreover, UBE2V1 facilitated the proliferation and migration of HCC cells. Further investigation revealed that up-regulated UBE2V1 competes with HIF-1α for binding to the β-domain of von Hippel–Lindau (VHL) protein and, in complex with UBE2S, catalyzes K11/K48-linked ubiquitination at VHL K196, leading to its proteasomal degradation. This disruption of VHL function attenuates HIF-1α ubiquitination and degradation, resulting in sustained HIF-1α stabilization, increased nuclear accumulation, and enhanced transcriptional activity. Consistent with these findings, genetic knockdown of UBE2V1 or pharmacological inhibition of HIF-1α markedly suppresses HCC tumorigenesis and metastasis in vivo. Altogether, our study unveils a previously unrecognized HIF-1α–UBE2V1 positive feedback loop that is self-reinforcing and critically sustains the hypoxic microenvironment to drive HCC progression, highlighting UBE2V1 as both a promising prognostic biomarker and a compelling therapeutic target for HCC.

Acute myeloid leukemia (AML) is a blood cancer characterized by uncontrolled growth of myeloid cells. Overcoming AML treatment resistance, particularly to anthracycline-based drugs like doxorubicin (ADR), poses a challenge. This study investigated the role of CELF1, an RNA-binding protein, in ADR resistance and autophagy regulation in AML. CELF1 expression was elevated in multiple tumor types, including AML. AML cell lines exhibit varying levels of CELF1 expression, with drug-resistant cell lines showing higher CELF1 expression compared to parental cells. CELF1 knockdown reduced drug resistance, promoted cell death, and inhibited autophagy. Mechanistic analysis identified ATG5 as a potential CELF1-regulated target gene, with CELF1 knockdown reducing ATG5 expression and mRNA decay. These findings indicate that targeting CELF1 could overcome ADR resistance in AML by modulating autophagy through ATG5 regulation, highlighting its clinical significance as a therapeutic target for enhancing ADR response in AML.

Background & Aims:Prothrombin induced by vitamin K absence-II (PIVKA-II) levels have been reported to correlate with hepatocellular carcinoma (HCC) prognosis, but its utility for assessing early treatment response remains underexplored. This study evaluated early PIVKA-II changes for predicting response and survival in HCC patients undergoing immune checkpoint inhibitors (ICIs) and targeted therapy. Methods:Eighty-two HCC patients were enrolled. Serum PIVKA-II levels were measured at baseline and after the first treatment cycle. Patients were stratified based on early PIVKA-II dynamics into a biochemical response group (≥50% reduction, n=40) and a non-response group (<50% reduction, n=42). Logistic regression and Cox proportional hazards models were used to identify predictors of objective response rate (ORR), progression-free survival (PFS), and overall survival (OS). Results:Time-dependent ROC analysis established ≥50% PIVKA-II decline as the early response threshold. The PIVKA-II response group had a significantly higher proportion of patients with Child-Pugh A, a lower incidence of extrahepatic metastasis, and significantly higher ORR (82.5% vs 38.1%, P<0.001). Median PFS and OS were not reached in the PIVKA-II responder group, compared to 8.9 months and 16.7 months, respectively, in the non-responder group (both P < 0.001). Multivariate analysis confirmed early PIVKA-II response as an independent predictor of PFS (HR=0.687, P<0.001) and OS (HR=0.709, P<0.001). Notably, in AFP-negative patients, an early PIVKA-II response was predictive of ORR and was associated with significantly longer PFS and OS. Conclusion:Early PIVKA-II response effectively predicts treatment response and prognosis in advanced HCC patients receiving ICI and targeted therapy, especially in AFP-negative patients.

BACKGROUND:Hepatocellular carcinoma (HCC) is a significant malignant tumor that is typically diagnosed late and has a poor prognosis. USP28 (Ubiquitin-specific protease 28), a deubiquitinating enzyme within the ubiquitin-specific proteases (USPs) family, plays a pivotal role in various biological processes, especially in cancer progression. However, its functions and molecular mechanisms in HCC are still unknown. METHODS:We first analyzed the expression level of USP28 in HCC tissues relative to normal tissues using TCGA database. This was further validated by qRT-PCR and Western Blot. To investigate the function of USP28 in HCC, CCK-8 assay, clone formation assay and Transwell assay were performed in control and USP28 knockdown or overexpressed HCC cells. To explore potential downstream targets of USP28, we used IP-MS analysis. The interaction between USP28 and KRT1 was confirmed by immunoprecipitation and immunofluorescence staining. Finally, we evaluated the in vivo effects of USP28 on HCC growth and metastasis using a ectopic tumor-bearing mouse model. RESULTS:The expression of USP28 in HCC tissues was significantly higher than that in normal tissues, and its high expression was associated with poor prognosis. Functional experiments showed that down-regulation of USP28 expression effectively inhibited the proliferation, migration and invasion of HCC cells, while overexpression of USP28 produced the opposite effect. Mechanistic investigations demonstrated that USP28 interacted with KRT1 and exerted deubiquitination on KRT1, thereby maintaining the stability of KRT1. Further studies revealed that USP28 knockdown resulted in decreased IFITM3 expression, which inhibited HCC cell proliferation. In addition, USP28 knockdown combined with sorafenib inhibited tumor growth and metastasis in tumor xenograft mice model. CONCLUSIONS:Our study confirmed the carcinogenic effects of USP28 by stabilizing KRT1 expression and promoting IFITM3. USP28 small molecule inhibitors can inhibit the proliferation of hepatocellular carcinoma cells and enhance the sensitivity of hepatocellular carcinoma cell lines to sorafenib. This provides a theoretical basis for USP28 to be a new clinical method to alleviate sorafenib resistance.

BACKGROUND:Hepatocellular carcinoma (HCC) is the sixth most prevalent cancer globally and the third leading cause of cancer-related mortality. Protein ubiquitination and deubiquitination play vital roles in human cancers. Ubiquitin-specific protease 13 (USP13) is a deubiquitinating enzyme (DUB) that is involved in many cellular processes. However, the mechanism by which USP13 regulates deubiquitination remains largely unknown. METHODS:Clinical data were analyzed via online databases. USP13 expression in HCC cell lines and tissues was analyzed via western blotting and immunohistochemistry. A lentivirus was used to established stable USP13-knockdown and USP13-overexpression cells. Cell Counting Kit-8, colony formation, wound healing, Transwell, and sphere formation assays were used to detect the malignant behaviors of HCC cells in vitro. A subcutaneous mouse model was used to investigate the function of USP13 in vivo. Co-immunoprecipitation, chromatin immunoprecipitation and dual-luciferase reporter assays were conducted to explore the molecular regulation. RESULTS:USP13 was upregulated in HCC cell lines and tissues, which predicted a poor prognosis in patients with HCC. Functional experiments in which USP13 was overexpressed or depleted revealed the oncogenic role of USP13 in driving HCC progression both in vitro and in vivo. Mechanistically, WW domain-containing ubiquitin E3 ligase 1 (WWP1) was identified as a binding protein of USP13. Furthermore, USP13 can interact with WWP1 and then remove the K29- and K48-linked polyubiquitination chains from WWP1 to stabilize the WWP1 protein via the ubiquitin-proteasome pathway. Moreover, Yin Yang 1 (YY1) was explored as a new transcription factor of USP13, and YY1 could also upregulate WWP1 expression through USP13. Moreover, YY1 and WWP1 were shown to participate in the oncogenic role of USP13. CONCLUSIONS:Our findings revealed the functional YY1/USP13/WWP1 signaling axis in HCC, identifying a promising therapeutic target for anti-HCC treatment.

Aims Early recurrence (ER) is strongly associated with poor long-term survival in patients with hepatocellular carcinoma (HCC). This study aimed to explore a prediction model based on machine learning (ML). Methods Six ML algorithms were constructed and compared. The top-performing model was further compared with a conventional logistic regression model. Model interpretability was assessed using SHapley Additive exPlanations (SHAP) values at both global and individual levels. Results Among 903 patients included, 351 (38.9%) experienced ER within two years. As a result, the random forest (RF) model was selected and demonstrated superior discrimination (AUC 0.917) compared with the logistic regression model (AUC 0.853). SHAP analysis identified multiple tumors, microvascular invasion, and tumor size > 5 cm as the key contributors to ER. An online calculator based on the RF model is accessible at: https://doctoryu.shinyapps.io/HCCEarlyRecurrencePredictor/. Conclusion The RF model offers a clinically interpretable and deployable tool to support individualized postoperative surveillance strategies.

Introduction Metastatic pancreatic neuroendocrine tumors (pNETs) carry a poor prognosis. Currently, no validated model exists to accurately predict survival in this population, highlighting the need for effective prognostic tools. Materials and methods In this study, we developed and validated a machine learning-based survival prediction model using data from the Surveillance, Epidemiology, and End Results (SEER) database. The model incorporated ten key prognostic factors, including AJCC TNM stage (T, N, M), tumor grade, primary surgery, non-primary site surgery, chemotherapy, primary site, age, and sex. The eXtreme Gradient Boosting (XGBoost) algorithm was applied to construct the model. Results A total of 1430 patients were included in the study. The XGBoost model showed strong predictive performance, with area under the receiver operating characteristic curve (AUROC) values of 0.781, 0.747, and 0.741 for 1-, 3-, and 5-year survival, respectively. The model was implemented in a web-based application that delivers real-time, individualized survival estimates to support clinical decision-making and personalized treatment planning. Conclusion By capturing complex interactions among prognostic variables, the model provides accurate survival predictions and supports patient-centered care. It addresses a critical gap in prognostic tools for metastatic pNETs.

Background: Despite the implementation of laparoscopic and robotic-assisted liver resection (LLR vs. RLR) in many centers, there remains controversy surrounding the differences in perioperative outcomes between the two approaches. This study aims to clarify the discrepancies in perioperative outcomes between LLR and RLR through a prospective study. Methods: Patients with HCC received LLR or RLR were included. The postoperative complications were categorized and evaluated employing the standardized Clavien-Dindo classification and the Comprehensive Complication Index (CCI) score. Specifically, the median CCI of 20.9 was set as the cut-off value for the occurrence of severe complications. A 1:2 propensity score matched (PSM) analysis was performed to control confounding bias. Results: A total of 273 patients were included, of whom 213 (78%) patients received LLR and 60 (22%) patients received RLR. After PSM, RLR was associated with a longer operative time but shorter hospital stays (all P < 0.05). Postoperative outcomes in terms of overall complications, major and minor complications, and mortality were similar between RLR and LLR groups (all P > 0.05). Of note, RLR is significantly associated with a lower CCI score, especially server complications (OR 0.826, 95%CI 0.386-0.883, P = 0.023). Conclusions: In terms of complication rates, RLR does not reduce the incidence of overall complications when compared to LLR, but it can reduce the severity of complications that occur. RLR, is a feasible and safe approach for patients with HCC.

Background and Aims:General transcription factor IIIC subunit 2 (GTF3C2) is one of the polymerase III transcription-related factors. Previous studies have revealed that GTF3C2 is involved in regulating cell proliferation. However, the role of GTF3C2 in hepatocellular carcinoma (HCC) remains unclear. This study aimed to determine its expression, biological function, and mechanism in HCC. Methods:The expression of GTF3C2 in HCC and non-tumor tissues, along with its clinical significance, was investigated using public databases and clinical samples. Reverse transcription-quantitative polymerase chain reaction and Western blot assays were performed to detect the expression of GTF3C2, ubiquitin specific peptidase 21 (USP21), mitogen-activated protein kinase 2 (MEK2), extracellular signal-regulated kinase 1/2 (ERK1/2), and p-ERK1/2 in cells. A luciferase reporter assay was conducted to explore the regulatory effect of GTF3C2 on USP21 transcription. Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine, and colony formation assays were performed to assess HCC cell proliferation. Subcutaneous injection of HCC cells into nude mice was used to evaluate tumor growth in vivo. Results:GTF3C2 expression was upregulated in HCC tissues and was positively correlated with advanced tumor stages and high tumor grades. HCC patients with high GTF3C2 expression had significantly worse survival outcomes. Knockdown of GTF3C2 suppressed the proliferation of Hep3B and HCCLM3 cells, while overexpression of GTF3C2 facilitated the proliferation of SNU449 and Huh7 cells. GTF3C2 promoted USP21 expression by activating its transcription, which subsequently increased the levels of MEK2 and p-ERK1/2 in HCC cells. Overexpression of both USP21 and MEK2 counteracted the GTF3C2 knockdown-induced inactivation of the ERK1/2 pathway. Moreover, GTF3C2 promoted HCC cell proliferation in vitro and tumor growth in vivo by regulating the USP21/MEK2/ERK1/2 pathway. Conclusions:Upregulation of GTF3C2 is frequently observed in HCC tissues and predicts poor prognosis. GTF3C2 promotes HCC cell proliferation via the USP21/MEK2/ERK1/2 pathway.

BACKGROUND:Aggressive recurrence (AR) is an important factor affecting prognosis after surgery for hepatocellular carcinoma (HCC). This study aimed to establish and evaluate a visual calculator to predict AR using by machine learning (ML) model. METHODS:Patients diagnosed with HCC at an early stage were reviewed. The prediction ability of each model was evaluated using accuracy, sensitivity, specificity, precision, F1 score, and the area under the curve (AUC). Then, the model's prediction performance was evaluated by calibration curves, decision curve analysis (DCA), and precision-recall curves (PRC). RESULTS:483 patients were ultimately included in this study. The baseline characteristics indicate that patients in the AR group exhibit poorer liver function and more advanced tumor features. Then, nine risk features were identified and incorporated into the nine development models, respectively. Among the models, the XGBoost model showed the best prediction ability (AUC 0.986, 95% CI: 0.983-0.988). Calibration curves, DCA, and PRC further demonstrated the robust performance and clinical applicability. Then, A web-based calculator was built. CONCLUSION:An explainable XGBoost model to predict AR for patients with early-stage HCC after surgery was feasible and effective, suggesting its superior potential in tailoring surgical strategies and optimizing personalized postoperative treatment plans.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality, necessitating novel therapeutic targets. This study explores the oncogenic role of integrin-linked kinase-associated phosphatase (ILKAP) in HCC and its underlying mechanisms. Database analyses (TCGA, UALCAN) revealed ILKAP overexpression in HCC, correlating with poor prognosis. Functional assays demonstrated that ILKAP knockdown significantly suppressed HCC cell proliferation and migration in vitro, while xenograft models confirmed its role in tumor growth in vivo. RNA sequencing identified 357 differentially expressed genes (DEGs), including 48 protein-coding DEGs, with glycolytic enzyme PGAM1 notably downregulated upon ILKAP silencing. ILKAP and PGAM1 expression were positively correlated in HCC tissues, and elevated PGAM1 levels were linked to worse survival. Notably, restoring PGAM1 in ILKAP-knockdown cells rescued proliferation and invasion, underscoring PGAM1’s critical role in ILKAP-mediated tumor progression. ILKAP depletion also reduced extracellular acidification rates and altered glycolysis-related gene expression, highlighting its role in metabolic reprogramming. These findings suggest that ILKAP drives HCC malignancy by modulating PGAM1 and glycolysis, providing a potential therapeutic target for HCC treatment. Further elucidation of the ILKAP-PGAM1 axis may offer new strategies for liver cancer management.