Xiaoling Luo

Cellular senescence has been regarded as a tumor-suppressive mechanism when occurring in cancer cells. However, when senescence occurs in immune cells, where it can exert opposing effects. Macrophages, one of crucial component of the immune cell have been shown to undergo senescence in mouse models of glioma and lung cancer. Yet, evidence for macrophage senescence in hepatocellular carcinoma (HCC) remains limited. To investigate the senescent state of macrophage in HCC we established an in vitro co-culture model of murine hepatocellular carcinoma cell line Hepa1-6 and macrophages-like transformed murine cell line RAW264.7. SA-β-Gal staining, real-time quantitative PCR, bulk RNA-seq, immunofluorescence were employed to demonstrate macrophage senescence which co-culture with hepatocellular carcinoma. Real-time qPCR, phagocytosis assays, flow cytometry, and transwell invasion experiments revealed the function of macrophages. RAW 264.7 cells exhibited morphological changes consistent with senescence-like change, along with elevated staining of the senescence-associated marker SA-β-Gal and higher proportion of G1 phase in Cell cycle analysis. Co-cultured RAW264.7 cells exhibited upregulated expression of senescence-related genes while downregulating proliferation and cell cycle-related genes. Additionally, senescent macrophages displayed impaired function and adopted an M2-like phenotype. In summary, our findings indicate that macrophages co-cultured with hepatocellular carcinoma cells acquire senescence-like phenotype and the function has also changed.

Disulfidptosis is a recently identified form of programmed cell death driven by NADPH metabolic imbalance and disulfide stress, but its therapeutic relevance in hepatocellular carcinoma (HCC) remains poorly understood. RRx-001, a clinical-stage small-molecule agent known for its epigenetic modulatory and radiosensitizing effects, has yet to be explored for its potential to induce disulfidptosis and immunogenic cell death (ICD). This study investigated the mechanism by which RRx-001 triggers disulfidptosis in HCC through NADPH metabolic dysregulation and evaluated its capacity to elicit ICD and antitumor immunity. Using Huh-7 and Hepa1-6 HCC cell lines and a murine subcutaneous xenograft model, we assessed drug sensitivity (CCK-8), apoptosis (flow cytometry), metabolic parameters (NADPH, GSH/GSSG, ROS), and ultrastructural changes (transmission electron microscopy). Protein expression was analyzed by immunofluorescence and Western blotting. In vivo antitumor efficacy was evaluated, and immune microenvironment dynamics were characterized via transcriptomic sequencing, immunohistochemistry, and flow cytometry, with all animal experiments randomized and blinded. RRx-001 markedly reduced NADPH levels by downregulating G6PD, leading to redox imbalance (decreased GSH/GSSG ratio, elevated ROS) and F-actin cytoskeletal contraction—hallmarks of disulfidptosis. This process was partially reversed by the disulfide reductant TCEP, confirming disulfidptosis dependency; quantitative F-actin fluorescence intensity showed significant contraction in RRx-001-treated cells that was mitigated by TCEP co-treatment ( p < 0.01). Additionally, RRx-001 promoted the release of damage-associated molecular patterns (DAMPs), including CRT, HMGB1, and HSP70/90, activating ICD, as confirmed by ELISA of extracellular HSP70/90. In vivo, RRx-001 significantly suppressed tumor growth ( p < 0.001), reduced tumor weight, enhanced infiltration of CD4+ and CD8+T cells, increased M1 macrophage polarization, and downregulated PD-L1 expression. Transcriptomic analysis implicated enhanced antitumor immunity via T-cell receptor signaling and T-helper cell differentiation pathways. These findings demonstrate that RRx-001 triggers disulfidptosis in HCC by targeting the G6PD–NADPH axis while concurrently inducing ICD, achieving dual metabolic and immunomodulatory effects. This mechanistic insight provides a scientific foundation for developing novel disulfidptosis-based HCC therapies with high translational potential and suggests that future studies should explore the synergistic efficacy of RRx-001 with immune checkpoint inhibitors.

Soluble vector family member 6 (SLC2A6) has been implicated in the aggressiveness and poor prognosis of various cancers, yet its specific role in hepatocellular carcinoma (HCC) remains to be fully elucidated. This study utilized multiple databases to investigate the relationship between SLC2A6 expression and clinical stage, methylation status, drug sensitivity, immune infiltration, and immune checkpoint regulation. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted. Furthermore, in vitro and in vivo experiments were performed to assess the impact of SLC2A6 knockout on the proliferation, migration, invasion, and underlying mechanisms in hepatocellular carcinoma (LIHC) cells. SLC2A6 expression was significantly correlated with tumor prognosis, clinical stage, and methylation levels, and was found to influence immune cell infiltration and immune checkpoint gene expression. In LIHC, SLC2A6 was associated with key biological processes, including the cell cycle, P53 signaling, and ferroptosis. Knockdown of SLC2A6 markedly suppressed the proliferation, migration, and invasion of HCC cells, with this inhibition being closely tied to the ferroptosis pathway. SLC2A6 plays a pivotal role in the regulation of pan-cancer processes, particularly in tumor prognosis and immune-related mechanisms. In LIHC, it emerges as a potential prognostic biomarker and therapeutic target for the regulation of ferroptosis, offering new insights for targeted cancer therapies.

Cell death is a normal physiological process within cells that involves multiple pathways, such as normal DNA damage, cell cycle arrest, and programmed cell death (PCD). Cell death has been a hot spot of research in tumor-related fields, especially programmed cell death, which is a key form of cell death and is classified into different types according to the mechanism of occurrence, such as apoptosis, autophagy, necroptosis, pyroptosis, ferroptosis, and disulfidptosis. Given the important role of PCD in maintaining tissue homeostasis and inhibiting tumorigenesis and development, more and more basic and clinical studies are devoted to revealing its potential application in anti-tumor strategies. The purpose of this review is to systematically review the regulatory mechanisms of PCD and to summarize the latest research progress of anti-tumor treatment strategies based on PCD.

Chlorogenic acid (CGA) has demonstrated anti-tumor effects across various cancers, but its role in cholangiocarcinoma (CCA) remains unclear. Our study revealed CGA’s potent anti-tumor effects on CCA, significantly suppressing cell proliferation, migration, colony formation, and invasion while inhibiting the epithelial–mesenchymal transition. CGA induced apoptosis, modulated cell cycle progression, and exhibited a stable binding affinity to AKR1B10 in CCA. AKR1B10 was highly expressed in RBE cells, and CGA treatment reduced AKR1B10 expression. Knocking out AKR1B10 inhibited the proliferation of RBE cells, whereas the overexpression of AKR1B10 promoted their proliferation. Additionally, CGA suppressed the proliferation of RBE cells with AKR1B10 overexpression. Mechanistically, AKR1B10 activated AKT, and CGA exerted its inhibitory effect by reducing AKR1B10 levels, thereby suppressing AKT activation. Furthermore, CGA facilitated the polarization of tumor-associated macrophages towards an anti-tumor phenotype and enhanced T-cell cytotoxicity. These findings underscore CGA’s potential as a promising therapeutic agent for CCA treatment.

In the treatment of unresectable advanced hepatocellular carcinoma (HCC), cisplatin is administered transhepatic arterially for local treatment, but the clinical application of cisplatin drugs is frequently hindered by the emergence of drug resistance. Kinesin family member 2C(KIF2C) has been shown as oncogene in a variety of tumors. Nevertheless, its effect on cisplatin sensitivity has yet to be ascertained. Herein, we aim to investigate the impact of the KIF2C gene on cisplatin sensitivity within HCC and the plausible underlying molecular mechanism. We examined the expression level of the KIF2C gene in HCC cells by real-time quantitative reverse transcription PCR and Western blot analysis, and analyzed bioinformatically by The Gene Expression Omnibus database and The Cancer Genome Atlas database. The KIF2C gene was silenced using the small interfering RNA technology, and its effect on cisplatin drug sensitivity in HCC cells was evaluated by flow cytometry, cell proliferation, cell migration, and invasion assays. Our results indicated that KIF2C was highly expressed in HCC cells. KIF2C silencing inhibits HCC cell proliferation, migration and invasion, promotes apoptosis, and keeps the cell cycle in G2 phase. In addition, KIF2C silencing enhanced the sensitivity of HCC cells to cisplatin. KIF2C silencing down-regulates the expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and mitogen-activated protein kinase 3 (MAPK3) proteins. In conclusion, KIF2C silencing amplifies the sensitivity of HCC cells to cisplatin by regulating the PI3K/AKT/MAPK signaling pathway. Consequently, targeting KIF2C shows great application potential as a strategy for enhancing the effectiveness of HCC treatment.

Background: Accurate preoperative estimation of liver function reserve is the key to the safety of hepatectomy. Recently, indocyanine green retention test at 15 minutes (ICG-R15) has been widely used to estimate hepatic function reserve in different liver diseases. The purpose of this research was to investigate the clinical value of ICG-R15 in predicting postoperative major complications and severe posthepatectomy liver failure (PHLF) in patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) subjected to hepatectomy. Methods: A total of 354 HBV-associated HCC patients who underwent hepatectomy were enrolled. The Child–Pugh, model for end-stage liver disease (MELD), albumin–bilirubin (ALBI) and ICG-R15 for assessing postoperative complications risk were compared using receiver operating characteristic (ROC) curve and decision curve analysis (DCA). Results: Postoperative major complications developed in 32 patients (9.1%) and severe PHLF developed in 57 (16.1%) patients. Multivariate analyses revealed that ICG-R15 were independent factors for predicting postoperative major complications and severe PHLF. ROC curve analyses and DCA plots showed that the predictive abilities of ICG-R15 for postoperative major complications and severe PHLF risk was significantly greater than Child–Pugh, MELD, and ALBI scores. Similar results were obtained by stratifying different background subgroups. Then, patients were divided into three different risk cohorts, emphasizing the significantly discrepancy between the incidence of postoperative major complications and severe PHLF. Conclusion: Compared with Child–Pugh, MELD and ALBI scores, ICG‐R15 revealed significantly advantages in predicting postoperative major complications and severe PHLF in HBV-related HCC patients subjected to liver resection.

Background: Systemic inflammatory response (SIR) plays a crucial role in every step of tumorigenesis and development. More recently, the fibrinogen-to-albumin ratio (FAR), an inflammation-based model, was suggested as a prognostic maker for various cancer patients. This research aimed to estimate the prognostic abilities of FAR, neutrophil–lymphocyte ratio (NLR), monocyte-lymphocyte ratio (MLR), platelet– lymphocyte ratio (PLR), and systemic immune–inflammation index (SII) in patients with hepatocellular carcinoma (HCC) subjected to curative hepatectomy. Methods: A total of 1,502 cases who underwent hepatectomy for HCC were included. The predictive performances of FAR, NLR, MLR, PLR and SII were assessed with regards to overall survival (OS) and disease-free survival (DFS). The area under the time-dependent receiver operating characteristic curve was used to compare prognostic performances. Results: Data revealed that FAR had higher predictive accuracy than other inflammation-based models and alpha-fetoprotein (AFP) in assessing OS and DFS. Indeed, the OS and DFS of patients with high FAR (> 8.9), differentiated by the optimal cut-off value of FAR, were remarkably reduced (p < 0.05 for OS and DFS). Multivariate Cox regression analyses identified that AFP, FAR, clinically significant portal hypertension, tumor size, Barcelona Clinical Liver Cancer staging system, major resection and blood loss were independent indicators for predicting OS and DFS. Furthermore, these patients could be classified according to their FAR into significantly different subgroups, regardless of AFP levels (p < 0.05 for DFS and OS). Similar results were obtained in other inflammation-based prognostic models. Conclusions: Compared with NLR, MLR, PLR, SII and AFP, FAR showed significant advantages in predicting survival of HCC patients subjected to liver resection.