Cancer cachexia (CC) is a multifactorial syndrome characterized by progressive skeletal muscle wasting, systemic inflammation, and metabolic dysregulation. Traditional Chinese medicine (TCM) contains numerous bioactive compounds with potential multitarget effects, but systematic strategies for identifying and experimentally evaluating candidate compounds for CC remain limited. This study aimed to identify and evaluate a TCM-derived candidate compound for CC-associated skeletal muscle wasting. Bibliometric analysis was used to identify frequently studied TCM formulas and core herbs in cachexia-related research, followed by network pharmacology analysis of candidate compounds and associated pathways. Kaempferol (KA) was then evaluated in A549 tumor-bearing mice by assessing tumor burden, cachexia-related phenotypes, skeletal muscle histology, untargeted metabolomics, inflammatory mediators, muscle degradation-related markers, autophagy-associated endpoints, and ultrastructure. A549-conditioned medium (A549-CM)-treated C2C12 myotubes were used for complementary in vitro evaluation under conditions with or without supplemental prostaglandin E2 (PGE2). KA was selected as a representative candidate based on its distribution among the core herbs, network associations, and reported pharmacological activities. In A549 tumor-bearing mice, high-dose KA (100 mg/kg/day) significantly reduced tumor weight and was accompanied by improvements in food intake, estimated tumor-free body weight, gastrocnemius weight, myofiber cross-sectional area, and forelimb grip strength compared with the model group (all P < 0.05). KA treatment was also associated with lower serum TNF-α, IL-6, and PGE2 concentrations; reduced skeletal muscle COX-2, Atrogin-1, and MuRF1 expression; lower LC3B-II/LC3B-I ratio and p62 protein level; and fewer autophagic vacuole-like structures (all P < 0.05). Untargeted metabolomics revealed KA-associated alterations in arachidonic acid metabolism, glycerophospholipids, amino acids, and energy-related metabolites. In A549-CM-treated C2C12 myotubes, KA modulated atrophy-related gene expression, autophagy-associated endpoints, and cytoplasmic vacuolation in both the absence and presence of supplemental PGE2 (P < 0.05). KA reduced tumor burden and was accompanied by improvements in cachexia-related phenotypes and skeletal muscle outcomes. The molecular, metabolic, and cell-based findings further associated KA treatment with lower systemic inflammatory mediator levels, reduced muscle protein degradation-related markers, and alterations in inflammatory lipid metabolism and autophagy-associated endpoints. These findings support further investigation of KA as a TCM-derived candidate for CC-associated skeletal muscle wasting. Future studies should distinguish its direct muscle-related effects from the systemic benefits associated with reduced tumor burden and clarify whether KA alters PGE2-related signaling and autophagic flux.
Kaposi’s Sarcoma-associated Herpesvirus (KSHV) is the etiologic agent of several human cancers, including Kaposi’s Sarcoma (KS), which are still lacking of treatment options. Members of the bromodomain and extra-terminal domain (BET) family, especially bromodomain-containing protein 4 (BRD4), play important roles in RNA polymerase II–mediated transcriptional regulation and are required for the expression of many tumor-driving oncogenes in various cancer cells. Therefore, BET proteins have become attractive targets for anticancer drug development. Our recent studies showed that KSHV-infected immortalized endothelial cells displayed strong resistance to BET inhibitors such as (+)-JQ1. In contrast, we found out MZ-1 and SIM-1, two of BRD4 PROTAC degraders, as effective inhibitors of cell growth in these cells. To develop PROTAC molecules with enhanced BRD4 degradation potency, we introduced an (S)-methyl group at the benzylic position of the VHL ligand in the VHL-based BRD4 PROTACs MZ-1 and SIM-1, thereby designing and generating MZ-1-ME and SIM-1-ME, respectively. In KS cellular model, MZ-1-ME and SIM-1-ME exhibited enhanced BRD4 degradation and anticancer activities compared with their parental compounds, which may represent promising PROTACs for development of BRD4-targeted therapies against KS.
The lung is a frequent site of secondary metastasis for various cancers, not only due to its extensive vasculature and lymphatic network but also because its immune microenvironment is highly susceptible to inflammatory modulation by external insults such as smoking, aging, infection, or chemotherapy. Recent evidence indicates that neutrophils play a pivotal role in this process. Upon activation by chronic inflammation and tumor-derived factors, neutrophils form neutrophil extracellular traps (NETs) and release a wide range of inflammatory mediators, which remodel the extracellular matrix (ECM) and activate signaling pathways, collectively disrupting the dormancy-maintaining niche and triggering disseminated tumor cells (DTCs) proliferation. Moreover, neutrophils cooperate with immunosuppressive stromal and immune components to establish a permissive microenvironment that facilitates DTC immune evasion and reactivation. These insights redefine neutrophils as key “dormancy releasers” rather than passive bystanders in tumor progression. Understanding the molecular and cellular mechanisms underlying neutrophil-induced dormancy escape offers novel therapeutic opportunities for preventing metastatic recurrence in the lung.
Fuzheng Yiai Formula (FZYAF) is a traditional Chinese medicine compound, and its key components, ursolic acid (UA) and Anhydroicaritin (AHI), have potential antitumor activity, but their mechanisms of action in Prostate cancer (PCa) remain unclear. Key components and targets of FZYAF were identified by network pharmacology, and the components were further validated by HPLC-MS analysis. PCa cells were treated with UA and AHI to assess their effects on cell proliferation, migration, and ferroptosis. The interaction of UA or AHI with TP53 was assessed by molecular docking and DARTS. In vivo, a PCa mouse model was established, and tumor tissues from FZYAF-treated mice were subjected to metabolomic analysis. Additionally, the anti-tumor effects of FZYAF, UA, and AHI, and their impact on ferroptosis and the TP53 pathway were evaluated. Network pharmacology and HPLC-MS analysis, UA and AHI were identified as the key active components of FZYAF, and TP53 was predicted as a core target. In vitro, UA and AHI inhibited PCa cell proliferation and migration and induced ferroptosis, with their combination showing enhanced efficacy compared to either agent alone. Mechanistically, UA and AHI were bound to and upregulated the TP53 protein. In vivo, FZYAF inhibited tumor growth and induced ferroptosis in PCa mice, which was associated with alterations in lipid metabolism-related pathways, as revealed. UA and AHI, key components of FZYAF, inhibit PCa progression by binding to and upregulating the TP53 protein to induce ferroptosis. This study elucidates the role of FZYAF in developing anti-PCa drugs.
Polycystic ovary syndrome (PCOS) is a highly prevalent endocrine and metabolic disease. This study aimed to investigate the synergistic role of immune dysregulation and ferroptosis during the pathogenesis of PCOS. Through integrating multiple Gene Expression Omnibus (GEO) datasets, differentially expressed genes (DEGs) were screened and subjected to functional enrichment analysis; weighted gene co‑expression network analysis combined with LASSO, RandomForest, and support vector machine‑recursive feature elimination algorithms were employed to identify key candidate genes; in vitro, human ovarian granulosa cells (KGN) were treated with dihydrotestosterone (DHT), NNMT expression was regulated via lentiviral vectors together with ferroptosis inhibitor intervention, and cell viability, cytotoxicity, and ferroptosis‑related indicators were examined; in vivo, a PCOS mouse model was established using dehydroepiandrosterone, and ovarian pathology, hormone levels, as well as molecular expression of the ferroptosis pathway were analyzed. DEGs in PCOS were significantly enriched in immune response and ferroptosis pathways. Twelve candidate genes were identified through screening multiple GEO datasets and demonstrated diagnostic potential. DHT induced ferroptosis in KGN cells by upregulating NNMT, manifesting as reduced cell viability, increased LDH release, accompanied by Fe²⁺ accumulation, enhanced lipid peroxidation, downregulation of SLC7A11/GPX4, and upregulation of ACSL4. This process could be partially reversed by ferroptosis inhibitors. Knockdown of NNMT effectively alleviated the DHT‑induced ferroptosis phenotype. Ovaries from PCOS mice exhibited typical pathological alterations, concurrent with elevated NNMT expression and abnormal activation of the ferroptosis pathway. This study has uncovered novel mechanisms underlying the development and progression of PCOS, offering potential targets for its diagnosis and treatment.
Nonalcoholic fatty liver disease (NAFLD) is an emerging health issue worldwide. It involves intrahepatic lipid accumulation that is potentially deleterious. Traditional Chinese Medicine called Huangjing Jiangzhi (HJJZ) granules holds promise for NAFLD treatment. This study explored the mechanism by which HJJZ granules improve NAFLD in male C57BL/6 mice and oleic acid-induced HepG2 cells. H E staining, Oil Red O staining, the TUNEL assay, and Western blotting were employed. Chromatin immunoprecipitation and dual-luciferase reporter assays were used to examine transcriptional regulation. HJJZ granules alleviated hepatic steatosis, reduced serum ALT/AST levels, improved lipid profiles, suppressed hepatocyte apoptosis, and inhibited the PERK-eIF2α-CHOP pathway in high-fat diet-fed mice and oleic acid-treated cells. These granules specifically upregulated the expression of the transcription factor FOXA1. FOXA1 knockdown abolished all protective effects of HJJZ granules. Mechanistically, FOXA1 was shown to transcriptionally activate DERL1 expression. DERL1 overexpression rescued the steatotic, apoptotic, and endoplasmic reticulum (ER) stress phenotypes in FOXA1-deficient cells. HJJZ granules ameliorate NAFLD by attenuating lipid accumulation, apoptosis, and ER stress. This protective effect is mediated through FOXA1 upregulation, which, in turn, transcriptionally enhances DERL1 expression, thereby mitigating ER stress. The FOXA1/DERL1 axis is a critical mechanistic pathway for HJJZ granule action.
Liver cirrhosis remains a challenging global health issue for which current treatment options are largely limited to managing acute liver failure, with no definitive curative therapies available. The Ruangan Huayu Formula (RGHY) has shown promising clinical effects in recent years; however, its comprehensive chemical profile and the specific mechanisms underlying its anti-cirrhotic activity remain to be fully elucidated. Ultra-high-performance liquid chromatography-Orbitrap-explorative mass spectrometry (UHPLC-OE-MS) was performed to characterize the chemical profile of the RGHY. Network pharmacology and molecular docking were leveraged to identify potential targets and pathways. Antifibrotic efficacy and underlying mechanisms of RGHY were evaluated using a thioacetamide (TAA)-induced liver fibrosis model in male Sprague-Dawley rats, along with transforming growth factor-beta (TGF-β)-stimulated human hepatic stellate cells (LX-2) in vitro. UHPLC-OE-MS study showed various chemical components of RGHY. Network pharmacology analysis revealed 57 intersecting targets, with the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway standing out. Molecular docking studies returned strong binding scores for AKT1, SRC, PIK3CA, and STAT3 as core targets key to RGHY compounds. RGHY was shown to significantly alleviate TAA-induced liver injury and fibrosis by down-regulating the PI3K/AKT pathway. In vitro, RGHY-containing serum and its key bioactive compounds (7,3’,4’-trihydroxyflavone and sulfuretin) disrupted TGF-β-induced LX-2 cell activation, migration, and fibrogenesis. The PI3K/AKT signaling pathway was confirmed to involve in mediating the antifibrotic effects of RGHY. RGHY and its bioactive compounds 7,3’,4’-trihydroxyflavone and effectively attenuated liver fibrosis and hepatic stellate cell activation through multicomponent, multitarget actions, primarily by modulating the PI3K/AKT pathway. These findings provide mechanistic evidence supporting its traditional use and its potential as a therapeutic candidate for liver cirrhosis.
Evasion of apoptosis is a major cause of therapy failure in lymphoma. We studied a covalent compound 6u with an α, β-saturated carbonyl group derived from the diuretic drug ethacrynic acid to induce apoptosis of lymphoma cells by targeting antiapoptotic proteins. We found that 6u induces apoptosis in 4 lymphoma cell lines, Daudi, Romas, Jeko-1 and Jurkat, accompanied by down-regulation of c-Flip and Mcl-1, as well as upregulation of Noxa. The down-regulation of Mcl-1 relies on Noxa induction and that the down-regulation of c-Flip is mediated by inhibiting protein synthesis pathway through covalent binding to mTOR complexes. Overexpression of c-Flip and silencing Noxa attenuate 6u-induced apoptosis. 6u modified with the saturated carbonyl structure loses the mTOR inhibition and apoptosis induction abilities. 6u exhibits potent anti-lymphoma effects in mantle cell lymphoma Jeko-1 xenografts without causing toxicity. 6u functions as an apoptosis inducer through the α, β-saturated carbonyl group to decrease c-Flip and to induce Noxa. 6u represents a new type of covalent agent for targeting lymphoma by inducing apoptosis.
Joint arthroplasty, replacement of damaged articular surfaces with a prosthetic component, is an increasingly common orthopaedic procedure. Despite the overall success of arthroplasty, complications such as periprosthetic joint infection remain a persistent problem with severe consequences. With increasing clinical demands, the need for a reliable, reproducible model for testing therapeutic strategies for arthroplasty complications is fundamental. In this study, a novel small animal model was designed and evaluated for downstream investigations. Two different implant designs (tibial vs. femoral) and surgical approaches were compared. The femoral implant resulted in fewer complications such as knee joint destabilization, severe swelling, and erosion of articular cartilage and subchondral bone. Rabbit care methodology was also optimized during model development, and both the process and results are presented to promote ethical and reproducible preclinical model research. The custom porous coated unicondylar femoral implant with a polished condylar surface demonstrated far fewer complications and adverse events than the tibial implant, as well as robust osseointegration. This will serve as a model for future investigations in preventing and treating joint arthroplasty complications. Joint arthroplasty, replacement of damaged articular surfaces with a prosthetic component, is an increasingly common orthopaedic procedure. Despite the overall success of arthroplasty, complications such as periprosthetic joint infection remain a persistent problem with severe consequences. With increasing clinical demands, the need for a reliable, reproducible model for testing therapeutic strategies for arthroplasty complications is fundamental. In this study, a novel small animal model was designed and evaluated for downstream investigations. Two different implant designs (tibial vs. femoral) and surgical approaches were compared. The femoral implant resulted in fewer complications such as knee joint destabilization, severe swelling, and erosion of articular cartilage and subchondral bone. Rabbit care methodology was also optimized during model development, and both the process and results are presented to promote ethical and reproducible preclinical model research. The custom porous coated unicondylar femoral implant with a polished condylar surface demonstrated far fewer complications and adverse events than the tibial implant, as well as robust osseointegration. This will serve as a model for future investigations in preventing and treating joint arthroplasty complications.
Hepatocellular carcinoma (HCC) is a highly lethal malignancy with high recurrence and limited therapies. Microvascular invasion (MVI) is a critical prognostic factor, but its molecular mechanisms and precise predictive tools are lacking. This study aimed to identify key MVI-associated genes (MVIRGs), develop a prognostic model, and elucidate the role of ANGPT2 in HCC progression and therapeutic responses. Machine learning on multi-cohort data (TCGA, ICGC) identified MVIRGs and constructed a prognostic model. ANGPT2, the core oncogenic MVIRG, was validated clinically (IHC) and functionally (in vitro and in vivo). We analyzed its association with the tumor immune microenvironment (TIME), immune checkpoint inhibitors (ICIs), and drug sensitivity. A powerful six-MVIRG prognostic model was developed, with ANGPT2 identified as the sole oncogenic gene. Clinically, ANGPT2 was significantly overexpressed, correlating strongly with aggressive features like MVI, vascular invasion, and advanced stages, serving as a robust poor prognostic biomarker for OS and PFS. Functionally, ANGPT2 knockdown significantly inhibited HCC cell growth and migration in vitro, and inhibited tumor growth in vivo. Bioinformatics revealed ANGPT2 fosters an immunosuppressive TIME, upregulating immune checkpoints (e.g., PD-L1) to mediate resistance to ICIs. High ANGPT2 expression showed a differential drug response: resistance to targeted therapies (e.g., EGFR-TKIs) but paradoxically increased sensitivity to chemotherapies (e.g., docetaxel, paclitaxel). This study constructed a powerful MVIRG model and identified ANGPT2 as a core oncogenic molecule. ANGPT2 drives HCC progression, mediates immune resistance, and differentially regulates drug sensitivity, making it a key biomarker for prognosis and treatment guidance.
Lipid metabolic reprogramming is increasingly recognized as a critical feature of prostate cancer progression, but the lipid metabolism-related genes that remain continuously dysregulated from normal tissue to primary tumor and metastatic disease have not been systematically characterized, and their biological and prognostic relevance remains incompletely understood. To identify lipid metabolism-related genes associated with continuous prostate cancer progression and develop a prognostic signature for survival stratification. Clinical prostate cancer specimens and a high-fat diet (HFD)-driven RM-1 tumor model were first used to evaluate lipid metabolic alterations in vivo. GSE6919 transcriptomic data were used to identify genes shared between the Normal–Primary and Primary–Metastatic transitions. These genes were intersected with a curated lipid metabolism-related gene set, followed by GO and KEGG enrichment analyses. TCGA prostate adenocarcinoma expression and clinical data were used for LASSO regression to construct a prognostic model. The four core genes were further evaluated by clinicopathological correlation analysis, protein- and transcript-level validation in clinical tissues and prostate cancer cell lines, and functional assays under oleic acid-induced lipid stress. Immune infiltration analysis, ssGSEA, and nomogram analysis were performed to assess the biological and clinical relevance of the model. Clinical tissues showed increased PLIN3 expression, and HFD feeding promoted tumor growth and reinforced lipid metabolic alterations in vivo. A total of 44 lipid metabolism-related genes were identified as continuously dysregulated during prostate cancer progression. These genes were mainly enriched in fatty acid metabolism, lipid catabolism, peroxisome, lipid droplet, glycolysis/gluconeogenesis, arachidonic acid metabolism, and PPAR signaling. Eight genes were significantly associated with overall survival in TCGA, and a four-gene signature comprising ALDH3A2, ENO2, PPP1CB, and PTGIS was established. This model effectively stratified patients into high- and low-risk groups with significantly different survival outcomes. The risk score was positively associated with clinical T stage and Gleason score. The four core genes were also associated with lipid metabolic enzymes, immune infiltration patterns, and multiple metabolism-related pathways. Protein- and transcript-level validation in clinical tissues and prostate cancer cell lines supported the biological relevance of the signature, although PTGIS showed a more context-dependent pattern. Functionally, silencing ENO2 reduced oleic acid-induced lipid peroxidation, whereas silencing PPP1CB enhanced it, while ALDH3A2 showed a more context-dependent effect. A nomogram integrating the risk score with clinical variables improved individualized survival prediction. We identified lipid metabolism-related genes continuously dysregulated during prostate cancer progression and established a four-gene prognostic signature with potential value for survival prediction and risk assessment. These findings highlight lipid metabolic rewiring as an important component of prostate cancer evolution and provide candidate biomarkers for future mechanistic and translational studies.
The oncogenic γ-herpesvirus known as the Kaposi’s sarcoma-associated herpesvirus (KSHV) is a principal causative agent of several cancers arising in patients with compromised immune systems. One of KSHV-related malignancies, primary effusion lymphoma (PEL), comprises transformed B cells harboring the viral episome and arises preferentially within the pleural or peritoneal cavities of patients including those with HIV infection. PEL is a rapidly progressing malignancy with a median survival time of several months even with conventional chemotherapy. One of the major reasons causing PEL chemotherapy failure is multidrug chemoresistance of lymphoma cells, however, there are only limited data exploring the mechanisms of PEL chemoresistance. In the current study, we demonstrate the role of hyaluronan (HA) signaling activation in rapamycin resistance in PEL cells, and that targeting HA signaling by small HA oligosaccharides (oHA) can augment rapamycin efficacy against PEL expansion in vivo. Our results provide the framework for the development of HA-targeted therapies clinical trials for PEL patients, which may represent promising adjuncts to reduce chemotherapy toxicity, improve treatment and prolong patients’ survival.
Nonalcoholic fatty liver disease (NAFLD) is a significant underlying driver of hepatocellular carcinoma; however, current clinical treatment options remain limited. Ginsenoside CK (CK), a natural bioactive compound, has shown promise in modulating lipid metabolism and protecting liver function. Nevertheless, its therapeutic potential against the pathogenesis of NAFLD and the associated molecular pathways is not fully understood. This study employed an integrated strategy that combines transcriptomic analysis with both in vivo and in vitro validation, utilizing a high-fat diet (HFD)-induced rat model to elucidate the efficacy and molecular mechanisms of CK in ameliorating NAFLD. An NAFLD rat model was established through HFD feeding to assess the effects of CK on various physiological and biochemical parameters, thereby clarifying its role in regulating lipid metabolism and providing hepatoprotection in vivo. Additionally, a free fatty acid (FFA)-induced Huh7 cell model was constructed to evaluate the in vitro activity of CK in promoting lipid metabolism and inhibiting lipid accumulation. Subsequently, transcriptomic analysis was conducted to identify potential targets and elucidate the mechanisms underlying the effects of CK. Finally, the proposed mechanism was verified through in vivo experiments. In vivo results indicated that CK treatment significantly mitigated the increases in body weight and organ indices induced by a HFD. Additionally, CK improved biochemical parameters in both liver and serum, alleviated liver injury, steatosis, and insulin resistance, thereby providing a protective effect on liver tissue. Furthermore, CK markedly reduced the deposition of intracellular lipid droplets in hepatocytes. It also enhanced the expression of fatty acid metabolism-related genes, CPT1 and CPT2, in the liver, while downregulating the expression of lipogenic genes, including ACC1, FAS, and SREBP1c. Mechanistic investigations revealed that CK modulates the PPARγ/CPT1A signaling axis, suppresses the expression of hepatic lipogenic genes, reduces lipid accumulation, and consequently improves insulin resistance and liver injury. Our study demonstrates that CK attenuates NAFLD progression by regulating the PPARγ/CPT1A signaling axis.
Abstract Background Radiotherapy (RT) is a cornerstone treatment for lung cancer brain metastasis (LCBM), yet acquired radioresistance frequently leads to recurrence. The molecular and metabolic mechanisms underlying this adaptive evolution at single-cell resolution remain poorly defined. Methods Patient-derived organoids (PDOs) from LCBM tissues were established to model clinical radiation responses. Paired pre- and post-RT samples underwent single-cell RNA sequencing (scRNA-seq) to delineate transcriptional, metabolic, and regulatory alterations associated with radioresistance. Results Single-cell analysis revealed substantial population remodeling following RT, characterized by depletion of proliferative cells and enrichment of a resilient Hypoxic-EMT subpopulation. Pseudotime analysis demonstrated lineage plasticity, showing a transition from proliferative to mesenchymal states. Mechanistically, a viral mimicry response involving NF-κB and STAT signaling supported stress adaptation. Resistant cells exhibited a hypermetabolic phenotype marked by metabolic plasticity, including hybrid bioenergetics coupling glycolysis with oxidative phosphorylation, enhanced lipid turnover via simultaneous fatty acid synthesis and degradation, and increased glutathione metabolism for reactive oxygen species buffering. Pharmacogenomic profiling indicated concurrent chemotherapy resistance but collateral sensitivity to PI3K/MEK inhibitors and epigenetic therapies. Conclusions These findings provide a high-resolution atlas of radioresistance in LCBM and suggest that targeting the Hypoxic-EMT niche or oxidative-antioxidant balance may overcome therapeutic resistance.
Background Oxidative stress arises from disrupted equilibrium between reactive oxygen species generation and cellular antioxidant capacity, serving as a pivotal mechanism in inflammatory pathology, particularly osteoarthritis development. The progression of osteoarthritis-related pain may additionally involve oxidative stress through its activation of nociceptive signaling pathways, but the mechanism of its influence on the progression of osteoarthritis is not yet unanimous. Methods The study utilized publicly available sequencing data, which included specimens from patients with osteoarthritis and matched healthy controls, retrieved from GEO database, and combined them after removing batch effects. A multi-step analytical approach was employed to identify pivotal diagnostic biomarkers for osteoarthritis. This strategy encompassed examining differential gene expression, constructing protein-protein interaction networks, applying machine learning algorithms, and assessing co-expression patterns. Subsequently, a diagnostic framework incorporating a nomogram was developed based on these candidate genes. To discern distinct patient subgroups, cluster analysis was conducted. Further investigations into the underlying biology revealed divergent functional pathways and immune cell infiltration landscapes across the identified subtypes, highlighting the putative roles of the key biomarkers. We investigated the effects of FKBP5 on osteoarthritis through both in vivo and in vitro experiments, examining chondrocyte proliferation capacity, apoptosis, cellular senescence, mitochondrial function, and cartilage degeneration. Result We successfully screened the key genes PPARGC1A, FKBP5, and MMP13 for the diagnosis of osteoarthritis, constructed a diagnostic model and a nomogram of osteoarthritis, and verified their excellent diagnostic ability. Based on these core genes we classified osteoarthritis into two subtypes. Enrichment analysis and immune infiltration analysis confirmed that FKBP5 regulates the immune microenvironment in osteoarthritis and influences the progression of osteoarthritis. Both in vivo and in vitro experiments confirmed that FKBP5 overexpression enhances chondrocyte viability, proliferative capacity, and mitochondrial function under inflammatory conditions, while reducing cellular senescence, apoptosis, and cartilage degeneration. FKBP5 may serve as a potential therapeutic target for osteoarthritis intervention. Conclusion Our study identified distinct molecular subtypes of osteoarthritis by analyzing genes associated with oxidative stress. Furthermore, we found that FKBP5 exerts a positive regulatory effect on chondrocyte function and mitigates cartilage degeneration in the osteoarthritis environment. This provides new insights for personalized treatment of osteoarthritis.
Non-small cell lung cancer (NSCLC) is the most prevalent subtype of lung cancer, with persistently low overall cure and survival rates, largely attributed to tumor immune escape. This study aims to explore the role of RBM15 in the immune escape of NSCLC and provide a potential therapeutic target. Firstly, RBM15, LncRNA EGFR-AS1, PCBP2, USP3, and PD-L1 were detected in NSCLC cells. Peripheral blood mononuclear cells (PBMCs) were co-cultured with NSCLC cells to assess PBMC cytotoxicity, CD8⁺T and CD107a+CD8+T cell proportion, and levels of IFN-γ, IL-10, and IL-2. To verify the mechanism, m6A enrichment on EGFR-AS1 was analyzed. The m6A modification sites on EGFR-AS1 were assayed. The interactions among EGFR-AS1-PCBP2, PCBP2-USP3, and USP3-PD-L1 were detected. We found that RBM15, EGFR-AS1 and PCBP2 were upregulated in NSCLC cells. Mechanistically, RBM15 stabilized EGFR-AS1 through m6A modification, increased the binding of EGFR-AS1 to PCBP2, and promoted USP3 expression. USP3 bound to PD-L1 and inhibited PD-L1 ubiquitination and degradation. In PBMC-NSCLC co-cultures, RBM15 downregulation increased PBMC cytotoxicity, CD8⁺T cell proportion, IFN-γ/IL-2 levels, and decreased IL-10 levels—effects partially reversed by EGFR-AS1 or USP3 overexpression. In conclusion, RBM15 enhances the immune escape of NSCLC through the EGFR-AS1/USP3/PD-L1 axis via m6A modification.
Background Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies with limited therapeutic options. Circular RNAs (circRNAs) have emerged as critical regulators of cancer progression; however, the functional role of hsa_circ_0003472 in PDAC remains unexplored. Methods Expression of hsa_circ_0003472 was assessed in 30 paired PDAC and adjacent normal tissues and pancreatic cancer cell lines using quantitative RT-PCR. Loss-of-function experiments were performed to evaluate effects on proliferation (CCK-8, EdU), apoptosis (TUNEL, Western blot), migration, and invasion (Transwell assays). Gemcitabine sensitivity was determined by IC50 analysis. Bioinformatic prediction and dual-luciferase reporter assays identified the downstream regulatory axis. Xenograft mouse models validated findings in vivo. Results hsa_circ_0003472 was significantly upregulated in PDAC tissues and cell lines. Silencing hsa_circ_0003472 inhibited proliferation, migration, and invasion while promoting apoptosis and enhancing gemcitabine sensitivity. Mechanistically, hsa_circ_0003472 functioned as a competing endogenous RNA by sponging miR-1253, thereby relieving suppression of excision repair cross-complementing group 1 (ERCC1). Rescue experiments confirmed that the oncogenic effects of hsa_circ_0003472 were mediated through the miR-1253/ERCC1 axis. In vivo, hsa_circ_0003472 knockdown significantly reduced tumor growth and recapitulated molecular changes observed in vitro. Conclusion hsa_circ_0003472 promotes PDAC progression and chemoresistance through the miR-1253/ERCC1 regulatory axis, representing a potential therapeutic target for this devastating malignancy.
Astragaloside IV (AS-IV), a key active component derived from the traditional Chinese medicinal plant Astragali, has been reported to exhibit various biological activities, including antioxidative, anti-inflammatory, immunoregulatory, and antineoplastic properties. This study aimed to elucidate the role of AS-IV in inhibiting gastric cancer (GC) growth, focusing on its impact on cell ferroptosis and the underlying molecular mechanisms. Proliferation and migration of GC cells upon AS-IV treatment were examined using CCK-8, colony formation, and Transwell assays. Ferroptosis induction was analyzed via ELISA, flow cytometry, and transmission electron microscopy. Ferroptosis suppressor protein 1 (FSP1) mRNA stability was assessed by the ActD assay, while RNA immunoprecipitation (RIP) was employed to confirm the interaction between FSP1 mRNA, Fat mass and obesity-associated protein (FTO) demethylase, and YTH N6-methyladenosine RNA-binding protein F2 (YTHDF2). The dual-luciferase reporter assay was used to explore FTO binding to N6-methyladenosine (m6A)-modified sites on FSP1 mRNA. Furthermore, AS-IV’s anti-tumor effects (20 mg/kg) were validated in vivo using gastric cancer xenograft and lung metastasis mouse models. AS-IV significantly suppressed the proliferation and migration of GC cells by inducing ferroptosis. Mechanistically, AS-IV down-regulated FTO, thus impairing its interaction with FSP1 mRNA and leading to increased m6A modification on FSP1 mRNA. This modification facilitated m6A recognition protein YTHDF2-mediated recognition and subsequent degradation of FSP1 mRNA. The reduction of FSP1 triggered ferroptosis, while the overexpression of FSP1 or inhibition of ferroptosis by ferrostatin-1 partially reversed AS-IV’s effects on cell viability and migration. In vivo, AS-IV effectively inhibited tumor growth and metastasis. This study highlights potent anti-GC effects of AS-IV, mediated by the suppression of FSP1 via the FTO/YTHDF2/m6A axis. The treatment of AS-IV inhibits the expression of demethylase FTO in GC cells, subsequently disrupting the binding between FTO protein and FSP1 mRNA. This disruption leads to a increase in the level of m6A modification on FSP1 mRNA, thereby enhancing the recognition and binding of m6A-recognizing protein YTHDF2, promoting the decay of FSP1 mRNA. Ultimately, the downregulation of FSP1 enhances ferroptosis in GC cells. Collectively. AS-IV holds great promise as a novel therapeutic strategy for gastric cancer treatment. This image was created using the Biorender website (https://www.biorender.com/).
ALKBH5, one of the RNA N6-methyladenosine (m6A) demethyltransferases, has been suggested to be involved in the progression of several cancers. The aim of this study was to investigate clinical significance and biological functions of ALKBH5 in promoting ovarian cancer progression. We found a significant upregulation of ALKBH5 expression in ovarian cancer tissues compared with normal tissues. Correlation analyses indicated an association between heightened ALKBH5 expression and FIGO stage, as well as lymph node metastasis. Importantly, increased ALKBH5 expression indicated shorter progression-free survival and overall survival. Moreover, we found that hypoxia induced an increase in ALKBH5 expression in ovarian cancer via an HIF-1α-dependent mechanism. Loss-of-function assays demonstrated that ALKBH5 knockdown inhibited ovarian cancer cell progression both in vitro and in vivo. Furthermore, we found that knockdown of ALKBH5-meidated m6A demethylation decreased Notch2 mRNA stability and expression, resulting in the inhibition of cell proliferation, invasion and metastasis in OC cells. In summary, our findings demonstrated that ALKBH5 promotes the progression of ovarian cancer by activating Notch2 signaling, and suggested that ALKBH5 functions as an oncogene and may serve as a prognostic biomarker and therapeutic target in ovarian cancer.
Gastric cancer remains a major cause of global cancer-related morbidity and mortality. Mouse models are indispensable tools for preclinical research into its mechanisms and therapies. Although Helicobacter pylori (H. pylori) infection is the primary risk factor for gastric cancer, developing mouse models based on this pathogen faces significant challenges. These include low bacterial colonization and survival rates, unpredictable and protracted tumorigenesis timelines, restrictions of host genetic backgrounds, the complexity of inflammatory and immune microenvironments, difficulties in standardized pathological assessment, and poor model reproducibility. In light of these limitations, research efforts have diversified into four principal categories of modeling approaches: chemical carcinogen-induced models, microbe-infected models (particularly those involving H. pylori), xenograft models, and genetically engineered mouse models. Each strategy offers distinct advantages and constraints, necessitating careful selection based on specific research objectives. This review comprehensively examines both conventional and emerging methods for establishing gastric cancer mouse models, situating them within a historical and evolving research landscape from past reliance on H. pylori to present and future approaches in the potential post-H. pylori infection era. We emphasize the applicability and compatibility of each modeling system with particular research goals, providing critical insights for selecting optimal in vivo platforms to advance the study of gastric carcinogenesis.
