Cell Biology and Toxicology最新文献

Long non-coding RNAs (lncRNAs) have been considered the main regulators of cancer progression through their regulation of diverse cellular processes. Autophagy, which exerts context-dependent dual effects on gastric cancer (GC), remains controversial, and its interplay with lncRNAs has yet to be fully elucidated. Through integrated in vitro and in vivo functional assessments, we illustrate that silencing Linc-ROR markedly inhibits GC cell proliferation, migration, invasion, and xenograft growth. Transmission electron microscopy and mRFP-GFP-LC3 dual-fluorescence reporters revealed that Linc-ROR overexpression suppresses autophagic flux, which was further confirmed by Western blot analysis. Mechanistically, Linc-ROR functions as a competing endogenous RNA (ceRNA) to sequester miR-145-5p, thereby upregulating CARMIL1 and activating ERK/mTOR signaling, leading to autophagy inhibition and promotion of GC cell growth and invasiveness. Notably, pharmacological inhibition of mTOR with Everolimus reversed these malignant phenotypes, highlighting a therapeutically actionable vulnerability. Clinically, serum exosomal Linc-ROR was significantly elevated in GC patients and outperformed carcinoembryonic antigen (CEA) in diagnostic accuracy. Collectively, our findings establish Linc-ROR as a master regulator of autophagy suppression and GC progression via the miR-145-5p/CARMIL1/ERK-mTOR axis, underscoring its potential as a therapeutic target, while serum exosomal Linc-ROR emerges as a promising noninvasive biomarker for GC.

Background: A disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1) is involved in the occurrence and development of myocardial fibrosis. Here, we sought to explore the specific regulatory mechanism of ADAMTS1 in cardiac fibrosis post-myocardial infarction (CFPMI).

Methods: Blood samples from patients with myocardial fibrosis were collected. A CFPMI mouse model and in vitro models involving human or mouse cardiac fibroblasts treated with TGF-β1 or Ang II were constructed. ChIP was used to confirm that SMAD2 binds to ADAMTS1, and Co-IP was used to verify the interaction between ADAMTS1 and HDAC6. Cellular models with SMAD2 knockdown, ADAMTS1 regulation, and HDAC6 inhibitor treatment were used to study their roles in fibrosis. Finally, AAV-shRNA-HDAC6 and ADAMTS1 inhibitor effects were verified in vivo.

Results: ADAMTS1 levels were higher in myocardial fibrosis patients' serum. Increased ADAMTS1 and p-SMAD2 were found in fibrotic mouse hearts and human cardiac fibroblasts stimulated with fibrotic factors. ChIP validated the binding of SMAD2 to ADAMTS1. Mechanistically, SMAD2 regulated ADAMTS1 expression during TGF-β1-induced fibrosis in human and mouse cardiac fibroblasts. Overexpression of ADAMTS1 enhanced the production of collagen fiber proteins in human and mouse cardiac fibroblasts induced by TGF-β1. Moreover, HDAC6 expression was elevated in CFPMI mouse hearts and ADAMTS1 inhibited HDAC6 to regulate fibrosis. ADAMTS1 interacted with HDAC6 during fibrosis. In vivo, shRNA-HDAC6 and ADAMTS1 inhibitor treatment alleviated myocardial fibrosis and improved cardiac function after CFPMI.

Conclusions: Targeting ADAMTS1/HDAC6 alleviated TGF-β1/SMAD2-associated cardiac fibrosis in CFPMI. This study may provide a novel theoretical basis for the treatment of myocardial fibrosis.

Objective and design: T lymphopenia is a common phenomenon in the sepsis patients, closely related with secondary infection and patients death, whereas its mechanism is still largely unelucidated. Hence, it is urgent to study the mechanism underlying sepsis induced T lymphopenia and lower morbidity and mortality of septic infection.

Material and methods: We performed Single-cell RNA-seq (scRNA-seq) and created an atlas of peripheral blood mononuclear cells (PBMCs) from sepsis patients and healthy volunteers (n = 3). To further illustrate mechanisms related to fundamental biological in sepsis, we investigated SNHG5 mediated T cell apoptosis.

Results: We collected blood samples of 3 sepsis patients and 3 healthy donors, separated PBMCs, and performed scRNA-seq to create a atlas of PBMCs. 7 cell clusters were identified and annotated, followed by differentially expressed genes and pathway analysis. Then, we systematically discussed the cellular heterogeneity, and generated gene expression patterns of 7 different cellular cluster in sepsis and healthy group. Further analysis DEG and further bioinformatics analysis of different cellular cluster indicated that SNHG5/miR-324-5p/CDK16 axis contributed to inflammatory T cell apoptosis in sepsis. And further inhibitory and functional experiments indicated that SNHG5/miR-324-5p/CDK16 axis contributed to inflammatory T cell apoptosis in sepsis.

Conclusion: This study unveiled molecular mechanisms related to T lymphopenia in sepsis, such as cell fate decisions and modulation of SNHG5/miR-324-5p/CDK16 axis, making SNHG5 a promising therapeutic measurement for sepsis.

Acalabrutinib is a frontline agent for chronic lymphocytic leukemia (CLL). The objective of this experiment was to screen 21 cardiovascular drugs with a focus on elucidating the metabolic inhibition of acalabrutinib by finerenone in vitro and in vivo. The ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was applied to quantify acalabrutinib and its major active metabolite ACP-5862 in liver microsomes in vitro and in rats in vivo. The half-maximal inhibitory concentrations (IC₅₀) of finerenone in rat liver microsomes (RLM), human liver microsomes (HLM) and recombinant human CYP3A4 (rCYP3A4) were 7.00, 21.29 and 3.52 μM, respectively. In RLM and rCYP3A4, finerenone inhibited the metabolism of acalabrutinib via mixed-type inhibition (competitive and non-competitive), with a non-competitive inhibition observed in HLM. Compared with the control group, in vivo pharmacokinetic analysis indicated that finerenone co-administration increased acalabrutinib exposure, as reflected by 4.08-, 7.16-, and 3.96-fold increases in AUC_(0-t), AUC_(0-∞), and half-life (t_1/2), respectively, accompanied by an 86.02% reduction in the clearance (CL_z/F). Its metabolite ACP-5862 demonstrated a 57.47% and 65.12% decrease in AUC_(0-t) and the maximum concentration (C_max), respectively, while CL_z/F was increased by 0.79-fold. Finally, acalabrutinib and finerenone were evaluated for binding to cytochrome P450 3A4 (CYP3A4) by molecular docking, yielding binding energies of -2.36 and -2.19 kcal/mol, respectively. Thus, in vivo and in vitro results consistently indicated that finerenone inhibited the metabolism of acalabrutinib, providing the basis for individualized dosing considerations.

Nickel nanoparticles (Ni NPs) are widely used in industrial and commercial sectors, raising concerns about their potential occupational and environmental toxicity. Male infertility has increased significantly in recent decades, with environmental exposures playing a recognized role. Ni NPs have been identified as toxic agents that induce testicular damage and sperm abnormalities, yet their underlying molecular mechanisms are unknown. In this study, mouse spermatogonia GC-1 cells were used as an in vitro model to explore the role of mitochondrial autophagy (mitophagy) in the induced apoptosis of Ni NPs. Ni NPs significantly reduced cell viability, increased intracellular ROS levels, disrupted mitochondrial membrane potential, and triggered germ cell apoptosis. PINK1 and Parkin, key mitophagy-related proteins, exhibited significant upregulation. Cyclosporin A was used to inhibit mitophagy, attenuating mitochondrial damage and reducing apoptosis. In addition, PINK1 knockdown achieved by lentiviral transfection confirmed its critical role in mediating Ni NPs-induced mitophagy and subsequent cell death. These findings demonstrate that overactivation of the PINK1/Parkin pathway promotes apoptosis to Ni NPs exposure by mitophagy. Our study provides new mechanistic insights into the role of mitophagy in reproductive damage caused by nanomaterials.

Ferroptosis is a novel form of programmed cell death that is distinct from apoptosis, necrosis, and autophagy (macroautophagy). It is characterized by alterations in intracellular iron levels and lipid peroxidation. Considering its close association with numerous diseases, the molecular mechanisms underlying cellular ferroptosis have recently emerged as a prominent research focus. Lipocalin 2 (LCN2) is a circulating protein involved in the regulation of diverse cellular processes in eukaryotes and is closely associated with ferroptosis. However, the modulation of ferroptosis by LCN2 exhibits significant tissue and disease specificity. This is not only attributed to the ability of LCN2 to mediate iron metabolism reprogramming, but also to its capacity to bidirectionally regulate oxidative stress and interact with multiple signaling pathways. Therefore, this review summarizes the mechanisms through which LCN2 contributes to ferroptosis and its tissue- and disease-specific regulatory functions. Additionally, the current status of the clinical translation of LCN2 as a biomarker and therapeutic target is explored.

Mycotoxins are fungal secondary metabolites widely detected in up to eighty percent of frequently consumed foods, strongly associated with toxicological mechanisms. Evidence indicates that hepatic pathophysiology entails gut microbiota dysbiosis mediated by the complex, bidirectional interactions within the gut-liver axis. This scoping review aims to provide insight into the relationship between mycotoxins, gut microbiota, and liver disease in animals, having been conducted following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines (n = 44). The analyzed species were hens, broilers, rabbits, mice, carps, turbots, Lateolabrax maculatus, chicks, sheep, and rats. The most altered liver parameters, as a consequence of mycotoxin exposure, were alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, malondialdehyde, reactive oxygen species, superoxide dismutase, glutathione peroxidase, tumor necrosis factor-α, lipopolysaccharide, and inflammatory infiltration. Gut microbiota changes were analyzed at phylum (Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Verrucomicrobia) and genus level (Bifidobacterium, Lactobacillus, Clostridium, Ruminococcus, Akkermansia, Escherichia, Allobaculum, Blautia, Staphylococcus, Prevotella, Bacteroides, Turicibacter, Corynebacterium, Roseburia, Coprococcus). What is more, out of more of 400 existing mycotoxins, only a small fraction of mycotoxins has been investigated in the interplay of the gut-liver axis ((Aflatoxin B1 (AFB1), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN), enniatins (ENNs) and T-2 toxin)). Therefore, more research is to better understand the interplay of interactions regarding mycotoxins and the gut microbiota-liver axis, focusing on the formulation of new functional foods and/or nutraceuticals as toxicity mitigating strategies.

Although thermal ablation has emerged as a minimally invasive and effective local treatment for hepatocellular carcinoma (HCC), its high postoperative recurrence rate remains a major clinical challenge. Sublethal heat stress can induce residual tumor cells to upregulate factors such as heat shock proteins (HSPs) and hypoxia-inducible factor-1α (HIF-1α), enhancing their survival tolerance. This process synergizes with components of the tumor microenvironment (TME), including myeloid-derived suppressor cells (MDSCs) and cancer-associated fibroblasts (CAFs), to collectively drive HCC recurrence. This article comprehensively reviews the research progress on the molecular mechanisms of tumor recurrence post-ablation, predictive biomarkers, and targeted therapeutic strategies. By deciphering multi-omics biomarkers, it provides new perspectives for predicting recurrence risk. Furthermore, this article also explores the potential of combination therapies, including targeting HSPs/HIF-1α, reversing immunosuppression, eliminating cancer stem cells (CSCs), and intervening in CAFs. This study provides a solid theoretical foundation for addressing the challenge of HCC recurrence, holding significant importance for improving patient prognosis and guiding clinical translation.

Background: Intervertebral disc degeneration (IVDD) acts as the prerequisite and pathological basis for a series of spinal degenerative diseases, and it remains one of the most harmful and difficult-to-treat conditions in this category. However, the exact pathogenesis of IVDD has not been fully elucidated.

Methods: Comprehensive analysis of data mining, bioinformatics and real-time quantitative PCR was used to pinpoint the key transcription factors participated in the progression of IVDD. The role of early growth response factor 1 (EGR1) in IVDD was determined through a series of loss- and gain-of-function experiments in vitro and in vivo. Mechanistically, bioinformatics, chromatin immunoprecipitation, and dual-luciferase reporter assays were applied to illustrate the interaction mechanism between microRNA-4306 (miR-4306) and Methionine adenosyltransferase 2A (MAT2A), or EGR1. Finally, rescue experiments were designed to assess the impact of the EGR1/miR-4306/MAT2A axis on nucleus pulposus cell function in vitro.

Results: EGR1 was highly expressed in degenerated nucleus pulposus tissues and lipopolysaccharide-induced nucleus pulposus cells, and expression levels of EGR1 were positively relevant with IVDD pathological grade. EGR1 overexpression aggravated lipopolysaccharide-induced pyroptosis and extracellular matrix degradation of nucleus pulposus cells, while EGR1 knockdown inhibited these effects in vitro and alleviated IVDD progression in mice in vivo. Mechanistically, EGR1 directly suppressed miR-4306 transcription by binding its promoter, and MAT2A was a target gene of miR-4306. Rescue experiments confirmed EGR1 knockdown inhibited lipopolysaccharide-induced nucleus pulposus cells damage by mediating the miR-4306/MAT2A axis.

Conclusion: This study suggested the EGR1/miR-4306/MAT2A axis played an important role in IVDD pathogenesis, which might be promising therapeutic targets for IVDD.

Neuroblastoma (NBL) is a pediatric malignancy with poor prognosis in high-risk cases. This study explores the function of albumin conformation factor 1 (ACF1) in NBL progression and delves into the underpinning mechanism. Exome and transcriptome sequencing were applied to analyze ACF1 mutations/expression in NBL tissues versus controls. ACF1 was knocked down in NBL cell lines (KELLY, BE2C, N2a) for in vitro assays (viability, proliferation, migration, apoptosis, therapy sensitivity) or in vivo xenograft/metastasis models with radiation/cisplatin. Mechanisms were probed via RNA-sequencing, chromatin immunoprecipitation, luciferase assays, co-immunoprecipitation, and immunofluorescence assays. Expression patterns and the correlations between ACF1, GCLM, and NAA20 were detected in human NBL tissue microarrays. ACF1 mutations and elevated expression correlated with advanced tumor staging, high-risk factors, and unfavorable prognosis in NBL datasets and TMAs. ACF1 knockdown suppressed NBL cell proliferation, mobility, and in vivo tumor growth/metastasis, while enhancing cisplatin/radiation sensitivity and apoptosis. Mechanistically, ACF1 knockdown reduced GCLM transcription via decreased H3K27ac/H3K4me3/Myc at its promoter, elevating lipid peroxidation and lowering glutathione (GSH) levels. Lactate induced ACF1 lactylation and nuclear translocation, promoted by NAA20 interaction (enhanced by lactate). NAA20 knockdown phenocopied ACF1 effects, rescued by GCLM overexpression. NAA20 and GCLM were upregulated in NBL datasets/TMAs. This study suggests that the NAA20-mediated ACF1 lactylation drives GCLM-dependent GSH synthesis, promoting NBL cell growth and metastasis. Targeting this axis may improve therapy response.