Multi-drug resistance is the major cause of chemotherapy failure in hepatocellular carcinoma (HCC). YAP, a critical effector of the Hippo pathway, has been shown to contribute to the progression, metastasis and invasion of cancers. However, the potential role of YAP in mediating drug resistance remains obscure. RT-qPCR and western blot were used to assess YAP expression in HCC cell lines. CCK-8 assays, flow cytometry, a xenograft tumour model, immunochemistry and GFP-mRFP-LC3 fusion proteins were utilized to evaluate the effect of YAP on multi-drug resistance, intracellular ROS production and the autophagy of HCC cells in vitro and in vivo. Autophagy inhibitor and rescue experiments were carried out to elucidate the mechanism by which YAP promotes chemoresistance in HCC cells. We found that BEL/FU, a typical HCC cell line with chemoresistance, exhibited overexpression of YAP. Moreover, the inhibition of YAP by shRNA or verteporfin conferred the sensitivity of BEL/FU cells to chemotherapeutic agents through autophagy-related cell death in vitro and in vivo. Mechanistically, YAP silencing significantly enhanced autophagic flux by increasing RAC1-driven ROS, which contributed to the inactivation of mTOR in HCC cells. In addition, the antagonist of autophagy reversed the enhanced effect of YAP silencing on cell death under treatment with chemotherapeutic agents. Our findings suggested that YAP upregulation endowed HCC cells with multi-drug resistance via the RAC1-ROS-mTOR pathway, resulting in the repression of autophagy-related cell death. The blockade of YAP may serve as a promising novel therapeutic strategy for overcoming chemoresistance in HCC.

Plant homeodomain finger protein 8 (PHF8) serves an activator of epithelial-mesenchymal transition (EMT) and is implicated in various tumors. However, little is known about PHF8 roles in hepatocellular carcinoma (HCC) and regulating E-cadherin expression. PHF8 expression pattern was investigated by informatic analysis and verified by RT-qPCR and immunochemistry in HCC tissues and cell lines. CCK8, xenograft tumor model, transwell assay, and tandem mCherry-GFP-LC3 fusion protein assay were utilized to assess the effects of PHF8 on proliferation, metastasis and autophagy of HCC cells in vitro and in vivo. ChIP, immunoblot analysis, rescue experiments and inhibitor treatment were used to clarify the mechanism by which PHF8 facilitated EMT, metastasis and autophagy. PHF8 upregulation was quite prevalent in HCC tissues and closely correlated with worse overall survival and disease-relapse free survival. Furthermore, PHF8-knockdown dramatically suppressed cell growth, migration, invasion and autophagy, and the expression of SNAI1, VIM, N-cadherin and FIP200, and increased E-cadherin level, while PHF8-overexpression led to the opposite results. Additionally, FIP200 augmentation reversed the inhibited effects of PHF8-siliencing on tumor migration, invasion and autophagy. Mechanistically, PHF8 was involved in transcriptionally regulating the expression of SNAI1, VIM and FIP200, rather than N-cadherin and E-cadherin. Noticeably, E-cadherin degradation could be accelerated by PHF8-mediated FIP200-dependent autophagy, a crucial pathway complementary to transcriptional repression of E-cadherin by SNAI1 activation. These findings suggested that PHF8 played an oncogenic role in facilitating FIP200-dependent autophagic degradation of E-cadherin, EMT and metastasis in HCC. PHF8 might be a promising target for prevention, treatment and prognostic prediction of HCC.

Hepatocellular carcinoma (HCC) is characterized by high morbidity and mortality, with limited effective treatment options. N ‐acetyltransferase 10 (NAT10) is the only known acetyltransferase for mRNA ac4C modification and is recognized as a biomarker for HCC, promoting its progression. However, the critical role of NAT10 in hepatocarcinogenesis remains to be fully elucidated, and the identification of suitable small‐molecule inhibitors targeting NAT10 is of great interest. Here, we report that NAT10 promotes HCC progression by stabilizing SMAD family member 3 (SMAD3) mRNA through ac4C modification. Clinically, NAT10 is highly expressed in HCC tissues and is significantly associated with poor prognosis. Functionally, NAT10 downregulation inhibits HCC cell proliferation, invasion, and epithelial‐mesenchymal transition, while promoting anoikis in vitro. Additionally, NAT10 depletion significantly impairs tumor growth, metastasis, and hepatocarcinogenesis in vivo. Mechanistically, NAT10 enhances oncogene SMAD3 mRNA stability via ac4C modification, thereby activating TGF‐β signaling pathway. We also identify a novel small‐molecule inhibitor, NAT10‐2023, which effectively blocks NAT10 activity. Notably, NAT10‐2023 treatment significantly reduces intracellular RNA ac4C modification levels and disrupts NAT10‐RNA interactions, leading to suppressed tumor progression. Overall, NAT10 drives HCC progression via SMAD3 mRNA stability regulation, and NAT10‐2023 could be a promising therapeutic candidate for targeting NAT10 in cancer treatment.

Atezolizumab has been widely used for the immunotherapy of various cancers. This study aims to analyze the adverse events (AEs) associated with Atezolizumab post-marketing using the FDA Adverse Event Reporting System (FAERS). We conducted a retrospective analysis of the FAERS database, focusing on reports associated with atezolizumab from the first quarter of 2016 to the third quarter of 2024. The ADRs were stratified by demographics, event types, and reported outcomes. We performed a disproportionality analysis using the reporting odds ratio (ROR), the proportional reporting ratio (PRR), the Bayesian confidence propagation neural network (BCPNN), and the empirical Bayesian geometric mean (EBGM) to assess the statistical association between atezolizumab use and specific AEs. Among the 18,310,937 non-duplicated reports collected from the FAERS database, 21,162 reports identified Atezolizumab as the “primary suspect (PS),” leading to the identification of 52,348 atezolizumab-induced AEs. The majority of AE reports (59.66%) occurred within the first 360 days of treatment, with most cases reported between 30 and 180 days of administration. A total of 494 Preferred Terms (PTs) were identified as significant across all four disproportionality-based pharmacovigilance signal assessment algorithms. The three system organ classes (SOCs) with the highest signal frequencies in AE reports were Gastrointestinal System Disorders (11.54%), Infectious and Invasive Disorders (10.12%), and Benign, Malignant, and Undetermined Tumors (including cystic and polypoid) (9.92%). The top five PTs with the strongest risk signals based on frequency screening were death, disease progression, pyrexia, anemia, and infectious pneumonia. Additionally, unexpected significant AEs were identified, including homicidal ideation, encephalitis, pancreatitis, disseminated intravascular coagulation, and Guillain- Barré syndrome. Atezolizumab treatment may be associated with a range of AEs. Further studies and continued pharmacovigilance efforts are essential to better understand the long-term safety of atezolizumab and to improve patient management strategies.

Glycogen synthase encoding genes GYS1 and GYS2 are aberrantly expressed and play a vital role in progression of multiple malignancies. However, the clinical significance and biological function of GYS1 and GYS2 in hepatocellular carcinoma remain little known. The present study aims to evaluate the relationship of GYS1 and GYS2 expression with clinicopathologic characteristics, prognostic pattern and immune infiltration in hepatocellular carcinoma (HCC) using the data from The Cancer Genome Atlas (TCGA). The expression of GYS1 and GYS2 in HCC and normal liver tissues were primarily compared using TCGA data and further validated by another independent cohort from the gene expression omnibus (GEO) database and human protein atlas (HPA). Wilcoxon rank sum test, Chi-square test and logistical regression analysis were used to examine the relationship of GYS1 and GYS2 expression with clinicopathological features. Gene ontology (GO) term analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene set enrichment analysis (GSEA) were conducted to explain the enrichment pathways and underlying molecular mechanism for GYS1 and GYS2. Kaplan-meier analysis, Cox regression and nomograms were used to evaluate the predictive value of GYS1 and GYS2 expression on clinical outcomes in HCC. Receive operating characteristic (ROC) curve was performed to illustrate the correlation of GYS1 and GYS2 expression with the risk of HCC. In addition, the relationship of GYS1 and GYS2 expression with immune infiltrate cells and immune checkpoint related genes were analyzed. Higher GYS1 expression and lower GYS2 expression were examined in HCC than in normal liver tissue and prominently associated with AFP level, tumor stage, histologic grade, pathologic stage and unfavorable overall survival (OS). ROC analysis showed that GYS1 and GYS2 may serve as reasonable diagnostic biomarkers for HCC patients. Moreover, GYS1 expression and GYS2 expression were significantly correlated with abundance of immune infiltrate cells and level of immune checkpoint related genes. Function annotation disclosed that both GYS1 and GYS2 were probably involved in synapse membrane, potassium ion transport, passive transmembrane transporter activity, channel activity, synaptic membrane, transport complex, neuroactive ligand-receptor interaction, allograft rejection, E2F targets, epithelial mesenchymal transition, G2M checkpoint and KRAS signaling pathway. The current study demonstrated that GYS1 upregulation and GYS2 downregulation might reshape tumor immune microenvironment and contribute progression of HCC. Both GYS1 and GYS2 were identified as promising biomarkers associated with diagnosis, prognosis and immune infiltration in HCC.