Cell Biology and Toxicology最新文献

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.

Ischemic stroke (IS) constitutes a leading driver of mortality and morbidity worldwide, with neuronal apoptosis representing a key pathological process. Accordingly, our objective was to delve into the implications of a novel signaling axis of the m6A methyltransferase METTL7B, lncRNA-MIR22HG, and JARID2 in driving neuronal apoptosis throughout cerebral ischemia/reperfusion (I/R) injury (CIRI). Mice subjected to middle cerebral artery occlusion/reperfusion (MCAO/R) and N2a cells exposed to oxygen-glucose deprivation/reoxygenation (OGD/R) were, respectively, established as in vivo and in vitro I/R models. METTL7B was markedly up-regulated after I/R and enhanced the m6A methylation and stability of lncRNA-MIR22HG, significantly prolonging its transcript half-life and amplifying its biological effects on neuronal fate. Stabilized lncRNA-MIR22HG suppressed the ubiquitin-mediated degradation of JARID2, thereby increasing JARID2 abundance and activity. Elevated JARID2 promoted the assembly of the p53/p300/MDM2 transcriptional complex, which in turn up-regulated the levels of pro-apoptotic genes, ultimately exacerbating neuronal apoptosis alongside ischemic brain injury. Functionally, METTL7B overexpression aggravated neurological deficits, infarct volume, and neuronal apoptosis in vivo, whereas METTL7B knockdown alleviated ischemic damage and conferred robust neuroprotection. Collectively, these findings define a novel METTL7B/lncRNA-MIR22HG/JARID2 signaling axis that integrates RNA methylation, lncRNA stabilization, proteostasis, and transcriptional activation of apoptosis, and highlight METTL7B as a potential novel target for therapeutic strategies aimed at preventing and treating ischemic stroke.

Parkinson's disease (PD) is characterized by dopaminergic neuronal loss, often associated with mitochondrial dysfunction and impaired mitophagy. Here, we investigated the role of HUWE1, an E3 ubiquitin ligase, in regulating mitophagy and neuronal survival in a cellular PD model. HUWE1 promoted mitophagy, whereas its depletion sensitized SH-SY5Y cells to 6-hydroxydopamine (6-OHDA)- and 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity and mitochondrial dysfunction. Notably, both toxins downregulated HUWE1, suggesting that loss of HUWE1 contributes to dopaminergic vulnerability. Conversely, HUWE1 overexpression preserved mitochondrial integrity and enhanced mitophagy under neurotoxic stress. Importantly, BL-918, a ULK1 activator that promotes AMBRA1 recruitment, facilitated HUWE1-mediated mitophagy in SH-SY5Y cells. BL-918 treatment significantly attenuated 6-OHDA- and MPP⁺-induced neurotoxicity and protected mitochondrial function via HUWE1 activation. Collectively, these findings identify HUWE1 as a key mechanistic regulator of mitophagy linked to dopaminergic neuronal vulnerability, and provide a conceptual framework for future investigations examining its role in PD-relevant model systems.

Osteoporosis, characterized by excessive osteoclast activity and bone resorption, is closely linked to mitochondrial respiration. The long non-coding RNA Gm5532 (Gm5532) has been implicated in osteoclast differentiation, but its role in mitochondrial function remains unclear. This study aimed to elucidate the mechanism by which Gm5532 regulates bone resorption through iron metabolism and mitochondrial respiration, focusing on its interaction with iASPP and the NRF2 signaling pathway. Here, we show that Gm5532 KO alleviates bone loss in aged, ovariectomized, and iron-overloaded mice by reducing osteoclast formation and activity. Mechanistically, Gm5532 directly interacts with the RNA-binding protein iASPP. This interaction modulates the KEAP1/NRF2 axis, leading to the destabilization of NRF2. Gm5532 KO enhances iASPP-KEAP1 binding, thereby stabilizing NRF2 and upregulating its target genes: Ftl, Fth, and Fpn1. This cascade reduces the intracellular labile iron pool. Iron deficiency suppresses mitochondrial biogenesis and respiration, and ultimately, inhibites osteoclast differentiation. In summary, Gm5532 functions as a critical regulator of bone resorption through its modulation of iron homeostasis and mitochondrial respiration. Our study uncovers a novel Gm5532-iASPP-NRF2 signaling axis that links iron metabolism to mitochondrial respiration and osteoclast function, offering a promising potential therapeutic target for osteoporosis.

Ferritinophagy, a selective autophagic process mediated by nuclear receptor coactivator 4 (NCOA4), plays a central role in maintaining cellular iron homeostasis by degrading ferritin and releasing stored iron. Under physiological conditions, this process dynamically regulates iron storage and utilization, thereby preventing both iron deficiency and iron overload. However, under pathological conditions such as chronic inflammation, oxidative stress, and harmful environmental exposures, aberrant activation of ferritinophagy leads to excessive ferritin degradation and abnormal expansion of the cytosolic labile iron pool. This, in turn, drives the accumulation of iron-dependent reactive oxygen species and lipid peroxidation, lowering the threshold for ferroptosis initiation, exacerbating tissue injury, and promoting disease progression. Thus, the pathological significance of ferritinophagy lies not only in iron mobilization itself but also in its close coupling with ferroptosis. This "ferritinophagy-ferroptosis axis" has emerged as a key framework for understanding the link between iron dysregulation and disease pathogenesis. In recent years, increasing evidence has shown that this axis is repeatedly activated in various chronic respiratory diseases (CRDs), where it exerts a pivotal role in disease onset and progression. This review systematically summarizes the molecular mechanisms of NCOA4-mediated ferritinophagy and highlights the potential pathogenic role of the ferritinophagy-ferroptosis axis in different CRDs, aiming to provide a theoretical basis for identifying novel therapeutic strategies and intervention targets.

Objective: The GTPase RhoA is known as a regulator involved in cartilage degeneration and subchondral bone remodeling related to osteoarthritis (OA). However, its specific role in synovial macrophages, the key immune cells of OA related tissues, remains entirely unexplored.

Methods: Herein, the RhoA expression in human and mouse OA synovium was analyzed. A macrophage-specific RhoA conditional knockout (cKO) mouse model was generated. Histological staining, OARSI scoring, and micro-CT were used to assess cartilage damage, while Western blot, immunofluorescence staining, and ELISA assessed changes in cellular function. Transcriptome sequencing and validation of signaling pathways were conducted using tissues and cells from patients with OA and OA mice.

Results: The collected results indicate that RhoA expression was significantly upregulated in synovial macrophages from OA patients and mice, correlating with disease severity. Contrary to its reported role in chondrocytes or endothelial cells, macrophage-specific RhoA deletion exacerbated OA, demonstrating enhanced cartilage destruction, subchondral bone loss, and synovitis. RhoA-deficient macrophages exhibited a pro-inflammatory M1 polarization and secreted high levels of IL-17C. This cytokine was necessary and sufficient to induce chondrocyte senescence, as evidenced by increased p53/p21, ROS, mitochondrial dysfunction, and suppressed autophagy, via activation of the PI3K/AKT/mTOR pathway. Mechanistically, RhoA ablation in macrophages activated the Hippo pathway effectors YAP/CCN2, leading to IL-17C transcription, independently of the canonical ROCK pathway.

Conclusion: In conclusion, present study reveals a previously unrecognized, protective role for macrophage RhoA in OA. It functions as a critical brake on a novel YAP-IL-17C axis, thereby preserving chondrocyte. This study redefines RhoA's role in joint homeostasis and nominates IL-17C as a potential therapeutic target for OA.

Repeated exposure to ketamine leads to mental behavioral disorders and cognitive deficits in mice. As a neurotransmitter receptor, dopamine receptor 1 (DRD1) is involved in mental regulation and memory formation. However, the role of DRD1 in ketamine's behavioral disorder and neurotoxicity remains unclear. We found that seven-day ketamine exposure induced anxiety-like, depressive-like behavior and cognition dysfunction in mice. DRD1 activation can produce anxiety-like behavior similar to that induced by ketamine. Furthermore, DRD1 activation synergistically exacerbates this effect of ketamine, and DRD1 antagonism partially attenuates the anxiety-like behavior and further aggravated the depressive-like behavior induced by ketamine. Moreover, ketamine induced HT22 cell apoptosis by DRD1 dependent inhibition of Akt/Gsk3β phosphorylation. DRD1 agonist synergistically enhanced the apoptosis induced by ketamine, while DRD1 antagonist or the apoptosis inhibitor partially reversed this apoptosis in vitro. In vivo assay found that ketamine promotes neuronal apoptosis in the hippocampus and prefrontal cortex of mice, and antagonizing DRD1 partially attenuates ketamine-induced apoptosis. In contrast, cell-specific knockdown of DRD1 in neuronal cells exacerbated ketamine-induced neuronal apoptosis and anxiety-like behavior. In summary, ketamine regulates DRD1 to suppress Akt/Gsk3β phosphorylation, inducing neuronal apoptosis, ultimately leading to anxiety-like behaviors in mice.

Engineered nanoparticles (ENPs), defined as nanoscale materials with at least one dimension between 1 and 100 nm, exhibit multifunctional and tunable physicochemical properties, that are at the center of several innovative fields. However, ENPs may induce a variety of biochemical reactions upon entry into organisms that could be a threat to human health. Therefore, a systematic evaluation of the toxicity of ENPs is essential. Quantitative structure-activity relationship (QSAR) is a practical in vitro modeling approach used to evaluate the toxicity of nanoparticles. In this study, we established the nanometric QSAR (Nano-QSAR) modelling based on cell membrane damage of ENPs to HepaRG cells. The toxicity data of ENPs and related 2D descriptor information were collected from the NanoCommons Knowledge Base. Periodic table descriptors of the elements were calculated using the Elemental Descriptor Calculator software. A multiple linear regression (MLR) model was constructed, and subsequently combined with read-across (RA) descriptors to establish the Nano-quantitative read-across structure-activity relationship (Nano-q-RASAR) model. Furthermore, machine learning (ML) algorithms were applied to optimize the predictive performance of the models. All models were validated according to the stringent OECD QSAR validation guidelines. Finally, a series of true external ENPs without experimental values were autonomously designed, and predicted using the best GB-Nano-QSAR model. Overall, this study can provide efficient and reliable predictions for the cell membrane damage of ENPs and a detailed theoretical explanation of their toxicity mechanism, which is of practical value for the toxicity assessment of ENPs.