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BACKGROUND: Metabolic reprogramming plays a pivotal role in cancer progression, contributing to substantial intratumour heterogeneity and influencing tumour behaviour. However, a systematic characterization of metabolic heterogeneity across multiple cancer types at the single-cell level remains limited. METHODS: We integrated 296 tumour and normal samples spanning six common cancer types to construct a single-cell compendium of metabolic gene expression profiles and identify cell type-specific metabolic properties and reprogramming patterns. A computational approach based on non-negative matrix factorization (NMF) was utilised to identify metabolic meta-programs (MMPs) showing intratumour heterogeneity. In-vitro cell experiments were conducted to confirm the associations between MMPs and chemotherapy resistance, as well as the function of key metabolic regulators. Survival analyses were performed to assess clinical relevance of cellular metabolic properties. FINDINGS: Our analysis revealed shared glycolysis upregulation and divergent regulation of citric acid cycle across different cell types. In malignant cells, we identified a colorectal cancer-specific MMP associated with resistance to the cuproptosis inducer elesclomol, validated through in-vitro cell experiments. Furthermore, our findings enabled the stratification of patients into distinct prognostic subtypes based on metabolic properties of specific cell types, such as myeloid cells. INTERPRETATION: This study presents a nuanced understanding of multilayered metabolic heterogeneity, offering valuable insights into potential personalized therapies targeting tumour metabolism. FUNDING: National Key Research and Development Program of China (2021YFA1300601). National Natural Science Foundation of China (key grants 82030081 and 81874235). The Shenzhen High-level Hospital Construction Fund and Shenzhen Basic Research Key Project (JCYJ20220818102811024). The Lam Chung Nin Foundation for Systems Biomedicine.
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Ergosterol peroxide (EP) isolated from the edible medicinal fungus Pleurotus ferulae has a wide range of anti-tumor activity, but poor water solubility and low bioavailability limit further application. In this study, EP was structurally modified using triphenylphosphine (TPP+), which combines mitochondrial targeting, amphiphilicity, and cytotoxicity. A series of TPP+-conjugated ergosterol peroxide derivatives (TEn) with different length linker arms were synthesized. The structure-activity relationship showed that the anticancer activity of TEn gradually decreased with the elongation of the linker arm. The compound TE3 has the optimal and broadest spectrum of antitumor effects. It mainly through targeting mitochondria, inducing ROS production, disrupting mitochondrial function, and activating mitochondria apoptosis pathway to exert anti-cervical cancer activity. Among them, TPP+ only acted as a mitochondrial targeting group, while EP containing peroxide bridge structure served as an active group to induce ROS. In vivo experiments have shown that TE3 has better anti-cervical cancer activity and safety than the first-line anticancer drug cisplatin, and can activate the immune response in mice. Although TE3 exhibits some acute toxicity, it is not significant at therapeutic doses. Therefore, TE3 has the potential for further development as an anti-cervical cancer drug.
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Antineoplásicos , Produtos Biológicos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Ergosterol , Mitocôndrias , Antineoplásicos/farmacologia , Antineoplásicos/química , Antineoplásicos/síntese química , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Humanos , Relação Estrutura-Atividade , Animais , Ergosterol/química , Ergosterol/farmacologia , Ergosterol/análogos & derivados , Camundongos , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Estrutura Molecular , Feminino , Apoptose/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/patologia , Linhagem Celular Tumoral , Pleurotus/química , Camundongos Endogâmicos BALB C , Compostos OrganofosforadosRESUMO
BACKGROUND: Interleukin (IL)-2 is a key cytokine capable of modulating the immune response by activating natural killer (NK) cells. This study was recruited to explore the therapeutic potential of IL-2-activated NK-92 cells in endometriosis in vitro. METHODS: Ectopic endometrial stromal cells (EESCs) were isolated and co-cultured with IL-2-activated NK-92 cells at varying effector-to-target (E:T) ratios (1:0 [Control], 1:1, 1:3, and 1:9). The viability, cytotoxicity, and cell surface antigen expression of IL-2-activated NK-92 cells were assessed. The viability, apoptosis, invasion, and migration ability of EESCs co-cultured with NK-92 cells at different ratios were evaluated. The apoptosis-related proteins, invasion and migration-related proteins as well as MEK/ERK pathway were examined via western blot. Each experiment was repeated three times. RESULTS: IL-2 activation enhanced NK-92 cytotoxicity in a concentration-dependent manner. Co-culturing EESCs with IL-2-activated NK-92 cells at E:T ratios of 1:1, 1:3, and 1:9 reduced EESC viability by 20%, 45%, and 70%, respectively, compared to the control group. Apoptosis rates in EESCs increased in correlation with the NK-92 cell proportion, with the highest rate observed at a 1:9 ratio. Moreover, EESC invasion and migration were significantly inhibited by IL-2-activated NK-92 cells, with a 60% reduction in invasion and a 50% decrease in migration at the 1:9 ratio. Besides, the MEK/ERK signalling pathway was down-regulated in EESCs by IL-2-activated NK-92 cells. CONCLUSION: IL-2-activated NK-92 cells exhibit potent cytotoxic effects against EESCs. They promote EESC apoptosis and inhibit viability, invasion, and migration through modulating the MEK/ERK signalling pathway.
Endometriosis is a common chronic systemic disease affecting approximately 190 million women worldwide. However, clinical treatments for endometriosis remain challenging due to the scarcity of high-quality scientific evidence and conflicting available guidelines. This research was designed to explore whether interleukin (IL)-2 affected the progression of endometriosis by modulating endometrial stromal cell apoptosis and natural killer (NK) cell-mediated cytotoxicity, thereby providing new therapeutic methods for endometriosis.
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Apoptose , Técnicas de Cocultura , Endometriose , Interleucina-2 , Células Matadoras Naturais , Humanos , Endometriose/patologia , Endometriose/imunologia , Feminino , Interleucina-2/farmacologia , Interleucina-2/metabolismo , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Apoptose/efeitos dos fármacos , Adulto , Endométrio/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Células Estromais/efeitos dos fármacos , Progressão da Doença , Sobrevivência Celular/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Células CultivadasRESUMO
Liver metastasis is common in patients with gallbladder cancer (GBC), imposing a significant challenge in clinical management and serving as a poor prognostic indicator. However, the mechanisms underlying liver metastasis remain largely unknown. Here, we report a crucial role of tyrosine aminotransferase (TAT) in liver metastasis of GBC. TAT is frequently up-regulated in GBC tissues. Increased TAT expression is associated with frequent liver metastasis and poor prognosis of GBC patients. Overexpression of TAT promotes GBC cell migration and invasion in vitro, as well as liver metastasis in vivo. TAT knockdown has the opposite effects. Intriguingly, TAT promotes liver metastasis of GBC by potentiating cardiolipin-dependent mitophagy. Mechanistically, TAT directly binds to cardiolipin and leads to cardiolipin externalization and subsequent mitophagy. Moreover, TRIM21 (Tripartite Motif Containing 21), an E3 ubiquitin ligase, interacts with TAT. The histine residues 336 and 338 at TRIM21 are essential for this binding. TRIM21 preferentially adds the lysine 63 (K63)-linked ubiquitin chains on TAT principally at K136. TRIM21-mediated TAT ubiquitination impairs its dimerization and mitochondrial location, subsequently inhibiting tumor invasion and migration of GBC cells. Therefore, our study identifies TAT as a novel driver of GBC liver metastasis, emphasizing its potential as a therapeutic target.
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Movimento Celular , Neoplasias da Vesícula Biliar , Neoplasias Hepáticas , Ribonucleoproteínas , Ubiquitinação , Animais , Humanos , Camundongos , Linhagem Celular Tumoral , Neoplasias da Vesícula Biliar/patologia , Neoplasias da Vesícula Biliar/genética , Neoplasias da Vesícula Biliar/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/secundário , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Camundongos Endogâmicos BALB C , Camundongos Nus , Mitofagia , Invasividade Neoplásica , Ribonucleoproteínas/metabolismo , Ribonucleoproteínas/genética , Tirosina TransaminaseRESUMO
The potential of human umbilical cord mesenchymal stromal cell-derived extracellular vesicles (hucMSC-EVs) in wound healing is promising, yet a comprehensive understanding of how fibroblasts and keratinocytes respond to this treatment remains limited. This study utilizes single-cell RNA sequencing (scRNA-seq) to investigate the impact of hucMSC-EVs on the cutaneous wound microenvironment in mice. Through rigorous single-cell analyses, we unveil the emergence of hucMSC-EV-induced hematopoietic fibroblasts and MMP13+ fibroblasts. Notably, MMP13+ fibroblasts exhibit fetal-like expressions of MMP13, MMP9, and HAS1, accompanied by heightened migrasome activity. Activation of MMP13+ fibroblasts is orchestrated by a distinctive PIEZO1-calcium-HIF1α-VEGF-MMP13 pathway, validated through murine models and dermal fibroblast assays. Organotypic culture assays further affirm that these activated fibroblasts induce keratinocyte migration via MMP13-LRP1 interactions. This study significantly contributes to our understanding of fibroblast heterogeneities as well as intercellular interactions in wound healing and identifies hucMSC-EV-induced hematopoietic fibroblasts as potential targets for reprogramming. The therapeutic targets presented by these fibroblasts offer exciting prospects for advancing wound healing strategies.
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Vesículas Extracelulares , Fibroblastos , Células-Tronco Mesenquimais , Análise de Célula Única , Cordão Umbilical , Cicatrização , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/química , Cordão Umbilical/citologia , Cordão Umbilical/metabolismo , Animais , Camundongos , Fibroblastos/metabolismo , Análise de Sequência de RNA , Células Cultivadas , Movimento Celular , Metaloproteinase 13 da Matriz/metabolismo , FetoRESUMO
OBJECTIVES: The purpose of this study is to determine the feasibility of polyetheretherketone-based dental implants, and analyze the stress and strain around different kinds of dental implants by finite element analysis. METHODS: The radiographic data was disposed to models in Mimics 19.0. 3D models of implants, crowns and jawbones were established and combined in SolidWorks 2018. Appling axial and oblique loads of 100 N, cloud pictures were exported in Ansys Workbench 18.0 to calculate and analyze the stress and strain in and around different implants. RESULTS: Oblique load tended to deliver more stress to bone tissue than axial load. The uniformity of stress distribution was the best for 30% short carbon fiber reinforced polyetheretherketone implants at axial and buccolingual directions. Stress shielding phenomenon occurred at the neck of 60% continuous carbon fiber reinforced polyetheretherketone and titanium implants. Stress concentration appeared in PEEK implants and the load of bone tissue would aggravate. CONCLUSIONS: 30% short carbon fiber reinforced polyetheretherketone implants demonstrate a more uniform stress distribution in bone-implant contact and surrounding bone than titanium. Stress shielding and stress concentration may be avoided in bone-implant interface and bone tissue. Bone disuse-atrophy may be inhibited in PEEK-based implants.
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Benzofenonas , Fibra de Carbono , Implantes Dentários , Prótese Dentária Fixada por Implante , Análise de Elementos Finitos , Cetonas , Polietilenoglicóis , Polímeros , Cetonas/química , Fibra de Carbono/química , Polímeros/química , Implantes Dentários/efeitos adversos , Polietilenoglicóis/química , Humanos , Prótese Dentária Fixada por Implante/efeitos adversos , Planejamento de Prótese Dentária , Análise do Estresse Dentário , Prótese Parcial Fixa , Carbono , Estresse Mecânico , Titânio/químicaRESUMO
OBJECTIVE: To explore the value of intravoxel incoherent motion (IVIM) and dynamic contrast enhanced MRI (DCE-MRI) for predicting phenotypic subtypes and Nottingham prognostic index (NPI) of breast cancer. STUDY DESIGN: Descriptive study. Place and Duration of the Study: Department of Radiology, Affiliated Hospital of Jining Medical University, Jining, Shandong, China, from March 2020 to January 2022. METHODOLOGY: One hundred and forty-one breast cancer patients with preoperative IVIM and DCE imaging were collected. IVIM parameters of D, D*, f, and DCE-MRI parameters of Ktrans, Kep, and Ve were measured. Receiver operating characteristic curves were conducted to assess the diagnostic efficacies. Additionally, 40 patients collected from February 2022 to July 2022 were enrolled as validation cohort. RESULTS: The D value in HER2-enriched (HER2-E) was lower than that in non-HER-E, while D*, Ktrans, and Ve values were higher than that in non-HER-E (p < 0.001, 0.046, < 0.001, and < 0.001, respectively). D + Ktrans + Ve showed an optimal diagnostic efficiency (AUC = 0.868). Meanwhile, D* and f values of triple-negative breast cancer (TNBC) were higher than those of non-TNBC, and Ve value of TNBC was lower than that of non-TNBC (p = 0.013, 0.006, and < 0.001, respectively). D* + f + Ve showed the best prediction performance (AUC = 0.849). Additionally, D and Kep were independent predictors of NPI (p < 0.001, and 0.002, respectively). D + Kep showed a good diagnostic efficiency (AUC = 0.818). CONCLUSION: The combined IVIM and DCE-MRI model showed enhanced diagnostic efficiency in predicting phenotypic subtypes and NPI of breast cancer, and might thus be considered efficient in therapy decision-making for patients. KEY WORDS: Breast neoplasms, Intravoxel incoherent motion, Dynamic contrast enhanced magnetic resonance imaging, Phenotypic subtypes, Nottingham prognostic index.
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Neoplasias da Mama , Neoplasias de Mama Triplo Negativas , Humanos , Feminino , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/patologia , Imagem de Difusão por Ressonância Magnética/métodos , Prognóstico , Meios de Contraste , Imageamento por Ressonância Magnética/métodosRESUMO
Marine permeable sediments are important sites for organic matter turnover in the coastal ocean. However, little is known about their role in trapping dissolved organic matter (DOM). Here, we examined DOM abundance and molecular compositions (9804 formulas identified) in subtidal permeable sediments along a near- to offshore gradient in the German North Sea. With the salinity increasing from 30.1 to 34.6 PSU, the DOM composition in bottom water shifts from relatively higher abundances of aromatic compounds to more highly unsaturated compounds. In the bulk sediment, DOM leached by ultrapure water (UPW) from the solid phase is 54 ± 20 times more abundant than DOM in porewater, with higher H/C ratios and a more terrigenous signature. With 0.5 M HCl, the amount of leached DOM (enriched in aromatic and oxygen-rich compounds) is doubled compared to UPW, mainly due to the dissolution of poorly crystalline Fe phases (e.g., ferrihydrite and Fe monosulfides). This suggests that poorly crystalline Fe phases promote DOM retention in permeable sediments, preferentially terrigenous, and aromatic fractions. Given the intense filtration of seawater through the permeable sediments, we posit that Fe can serve as an important intermediate storage for terrigenous organic matter and potentially accelerate organic matter burial in the coastal ocean.
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Matéria Orgânica Dissolvida , Ferro , Ferro/química , Água do Mar/química , Água , Compostos OrgânicosRESUMO
This study aims to explore the mechanism of Liujunzi Decoction in the treatment of 4-nitroquinoline-N-oxide(4NQO)-induced esophageal cancer in mice. One hundred mice of 35-45 days were randomized into blank, model, and low-, medium-, and high-concentration(18.2, 36.4, and 54.6 g·kg~(-1), respectively) Liujunzi Decoction groups. The mice in other groups except the blank group had free access to the water containing 100 µg·mL~(-1) 4NQO for 16 weeks for the modeling of esophageal cancer. The mice in the Liujunzi Decoction groups were fed with the diets supplemented with corresponding concentrations of Liujunzi Decoction. The body weight and organ weights were weighed for the calculation of organ indexes. The pathological changes of the esophageal tissue were observed by hematoxylin-eosin(HE) staining. Ultra performance liquid chromatography-mass spectrometry(UPLC-MS/MS) was employed to collect metabolites from mouse serum samples, screen out potential biomarkers, and predict related metabolic pathways. Compared with the blank group, the model group showed decreased spleen and stomach indexes and increased lung, esophagus, and kidney indexes. Compared with the model group, Liujunzi Decoction groups had no significant changes in the organ indexes. The HE staining results showed that Liujunzi Decoction inhibited the invasive growth and cancerization of the esophageal cancer cells. A total of 9 potential biomarkers of Liujunzi Decoction in treating esophageal cancer were screened out in this study, which were urocanic acid, 1-oleoylglycerophosphoserine, 11-deoxy prostaglandin E1, Leu-Glu-Lys-Glu,(±) 4-hydroxy-5E,7Z,10Z,13Z,16Z,19Z-docosahexaenoic acid, ureidosuccinic acid,(2R)-2,4-dihydroxy-3,3-dimethylbutanoic acid, kynurenic acid, and bicyclo prostaglandin E2, which were mainly involved in histidine, pyrimidine, alanine, aspartate, glutamate, pantothenate and tryptophan metabolism and coenzyme A biosynthesis. In summary, Liujunzi Decoction may exert the therapeutic effect on the 4NQO-induced esophageal cancer in mice by regu-lating the amino acid metabolism, inflammation, and immune function.
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Medicamentos de Ervas Chinesas , Neoplasias Esofágicas , Espectrometria de Massas em Tandem , Camundongos , Animais , Cromatografia Líquida , Metabolômica , Biomarcadores , Neoplasias Esofágicas/induzido quimicamente , Neoplasias Esofágicas/tratamento farmacológicoRESUMO
BACKGROUND: Microglia-mediated neuroinflammation in Alzheimer's disease (AD) is not only a response to pathophysiological events, but also plays a causative role in neurodegeneration. Cytoplasmic cysteinyl-tRNA synthetase (CARS) is considered to be a stimulant for immune responses to diseases; however, it remains unknown whether CARS is involved in the pathogenesis of AD. METHODS: Postmortem human temporal cortical tissues at different Braak stages and AD patient-derived serum samples were used to investigate the changes of CARS levels in AD by immunocytochemical staining, real-time PCR, western blotting and ELISA. After that, C57BL/6J and APP/PS1 transgenic mice and BV-2 cell line were used to explore the role of CARS protein in memory and neuroinflammation, as well as the underlying mechanisms. Finally, the associations of morphological features among CARS protein, microglia and dense-core plaques were examined by immunocytochemical staining. RESULTS: A positive correlation was found between aging and the intensity of CARS immunoreactivity in the temporal cortex. Both protein and mRNA levels of CARS were increased in the temporal cortex of AD patients. Immunocytochemical staining revealed increased CARS immunoreactivity in neurons of the temporal cortex in AD patients. Moreover, overexpression of CARS in hippocampal neurons induced and aggravated cognitive dysfunction in C57BL/6J and APP/PS1 mice, respectively, accompanied by activation of microglia and the TLR2/MyD88 signaling pathway as well as upregulation of proinflammatory cytokines. In vitro experiments showed that CARS treatment facilitated the production of proinflammatory cytokines and the activation of the TLR2/MyD88 signaling pathway of BV-2 cells. The accumulation of CARS protein occurred within dense-core Aß plaques accompanied by recruitment of ameboid microglia. Significant upregulation of TLR2/MyD88 proteins was also observed in the temporal cortex of AD. CONCLUSIONS: The findings suggest that the neuronal CARS drives neuroinflammation and induces memory deficits, which might be involved in the pathogenesis of AD.
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Doença de Alzheimer , Humanos , Animais , Camundongos , Camundongos Endogâmicos C57BL , Doença de Alzheimer/complicações , Doença de Alzheimer/genética , Fator 88 de Diferenciação Mieloide , Doenças Neuroinflamatórias , Receptor 2 Toll-Like , Proteínas Adaptadoras de Transdução de Sinal , CitocinasRESUMO
INTRODUCTION: Breast cancer (BC) is the most prevalent type of cancer and has the highest mortality among women worldwide. BC patients have a high risk of depression, which has been recognized as an independent factor in the progression of BC. However, the potential mechanism has not been clearly demonstrated. METHODS: To explore the correlation and mechanism between depression and BC progression, we induced depression and tumor in BC mouse models. Depression was induced via chronic unpredictable mild stress (CUMS) and chronic restraint stress (CRS). Amino acid (AA) neurotransmitter-targeted metabonomics and gut microbiota 16S rDNA gene sequencing were employed in the mouse model after evaluation with behavioral tests and pathological analysis. RESULTS: The tumors in cancer-depression (CD) mice grew faster than those in cancer (CA) mice, and lung metastasis was observed in CD mice. Metabonomics revealed that the neurotransmitters and plasma AAs in CD mice were dysregulated, namely the tyrosine and tryptophan pathways and monoamine neurotransmitters in the brain. Gut microbiota analysis displayed an increased ratio of Firmicutes/Bacteroides. In detail, the abundance of f_Lachnospiraceae and s_Lachnospiraceae increased, whereas the abundance of o_Bacteroidales and s_Bacteroides_caecimuris decreased. Moreover, the gut microbiota was more closely associated with AA neurotransmitters than with plasma AA. CONCLUSION: Depression promoted the progression of BC by modulating the abundance of s_Lachnospiraceae and s_Bacteroides_caecimuris, which affected the metabolism of monoamine neurotransmitters in the brain and AA in the blood.
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Aminoácidos , Neoplasias da Mama , Depressão , Progressão da Doença , Microbioma Gastrointestinal , Neurotransmissores , Animais , Microbioma Gastrointestinal/fisiologia , Feminino , Camundongos , Neurotransmissores/metabolismo , Aminoácidos/metabolismo , Depressão/metabolismo , Depressão/microbiologia , Neoplasias da Mama/patologia , Neoplasias da Mama/microbiologia , Neoplasias da Mama/metabolismo , Metabolômica , Modelos Animais de Doenças , Estresse Psicológico/microbiologia , Estresse Psicológico/metabolismo , Estresse Psicológico/complicaçõesRESUMO
PURPOSE: The detrimental effects of pathological angiogenesis on the visual function are indisputable. Within a prominent role in chromosome segregation and tumor progression, aurora kinase B (AURKB) assumes a prominent role. However, its role in pathological retinal angiogenesis remains unclear. This study explores this latent mechanism. METHODS: To inhibit AURKB expression, we designed specific small interfering RNAs targeting AURKB and transfected them into vascular endothelial cells. Barasertib was selected as the AURKB inhibitor. The anti-angiogenic effects of both AURKB siRNA and barasertib were assessed in vitro by cell proliferation, transwell migration, and tube formation. To evaluate the angiogentic effects of AURKB in vivo, neonatal mice were exposed to 75% oxygen followed by normoxic repositioning to establish an oxygen-induced retinopathy (OIR) model. Subsequently, phosphate-buffered saline and barasertib were administered into OIR mice via intravitreal injection. The effects of AURKB on cell cycle proteins were determined by western blot analysis. RESULTS: We found that AURKB was overexpressed during pathological angiogenesis. AURKB siRNA and barasertib significantly inhibited endothelial cell proliferation, migration, and tube formation in vitro. Furthermore, AURKB inhibition attenuated retinal angiogenesis in the OIR model. A possible mechanism is the disruption of cell cycle by AURKB inhibition. CONCLUSION: In conclusion, AURKB significantly influenced pathological retinal angiogenesis, thereby presenting a promising therapeutic target in ocular neovascular diseases.
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Organofosfatos , Quinazolinas , Doenças Retinianas , Neovascularização Retiniana , Animais , Camundongos , Angiogênese , Aurora Quinase B/antagonistas & inibidores , Aurora Quinase B/metabolismo , Divisão Celular , Proliferação de Células , Células Endoteliais/metabolismo , Camundongos Endogâmicos C57BL , Neovascularização Patológica , Oxigênio , Neovascularização Retiniana/metabolismo , RNA Interferente Pequeno/uso terapêuticoRESUMO
Hypertension is characterized by endothelial dysfunction and arterial stiffness, which contribute to the pathogenesis of atherosclerotic cardiovascular diseases. Nicotinamide adenine dinucleotide (NAD+) is an indispensable cofactor in all living cells that is involved in fundamental biological processes. However, in hypertensive patients, alterations in NAD+ levels and their relation with blood pressure (BP) elevation and vascular damage have not yet been studied. Here we reported that hypertensive patients exhibited lower NAD+ levels, as detected by high-performance liquid chromatography-mass spectrometry (HPLC-MS), in both peripheral blood mononuclear cells (PBMCs) and aortas, which was parallel to vascular dysfunction. NAD+ boosting therapy with nicotinamide mononucleotide (NMN) supplement reduced BP and ameliorated vascular dysfunction in hypertensive patients (NCT04903210) and AngII-induced hypertensive mice. Upregulation of CD38 in endothelial cells led to endothelial NAD+ exhaustion by reducing NMN bioavailability. Pro-inflammatory macrophages infiltration and increase in IL-1ß generation derived from pro-inflammatory macrophages resulted in higher CD38 expression by activating JAK1-STAT1 signaling pathway. CD38 KO, CD38 inhibitors treatment, or adeno-associated virus (AAV)-mediated endothelial CD38 knockdown lowered BP and improved vascular dysfunction in AngII-induced hypertensive mice. The present study demonstrated for the first time that endothelial CD38 activation and subsequently accelerated NAD+ degradation due to enhanced macrophage-derived IL-1ß production was responsible for BP elevation and vascular damage in hypertension. NAD+ boosting therapy can be used as a novel therapeutic strategy for the management of hypertensive patients.
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Hipertensão , NAD , Animais , Camundongos , Pressão Sanguínea , Células Endoteliais , Hipertensão/genética , Leucócitos Mononucleares , Regulação para Cima/genética , HumanosRESUMO
Autophagy and apoptosis are essential processes that participate in cell death and maintain cellular homeostasis. Dysregulation of these biological processes results in the development of diseases, including cancers. Therefore, targeting the interaction between apoptosis and autophagy offers a potential strategy for cancer therapy. Melanoma is the most lethal skin cancer. We previously found that tumor necrosis factor receptor-associated factor 6 (TRAF6) is overexpressed in melanoma and benefits the malignant phenotype of melanoma cells. Additionally, TRAF6 promotes the activation of cancer-associated fibroblasts in melanoma. However, the role of TRAF6 in autophagy and apoptosis remains unclear. In this study, we found that knockdown of TRAF6 induced both apoptosis and autophagy in melanoma cells. Transcriptomic data and real-time PCR analysis demonstrated reduced expression of autophagy related 16 like 2 (ATG16L2) in TRAF6-deficient melanoma cells. ATG16L2 knockdown resulted in increased autophagy and apoptosis. Mechanism studies confirmed that TRAF6 regulated ATG16L2 expression through c-Jun. Importantly, targeting TRAF6 with cinchonine, a TRAF6 inhibitor, effectively suppressed the growth of melanoma cells by inducing autophagy and apoptosis through the TRAF6/c-Jun/ATG16L2 signaling pathway. These findings highlight the pivotal role of TRAF6 in regulating autophagy and apoptosis in melanoma, emphasizing its significance as a novel therapeutic target for melanoma treatment.
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Laryngeal squamous cell carcinoma (LSCC) is the most common and prevalent malignant tumor in head and neck squamous cell carcinoma. Recent studies have shown that circular RNAs (circRNAs) play a vital role in cancer development, but their specific role in the tumorigenesis and development of LSCC remains unclear. We selected five pairs of LSCC tumor tissues and paracancerous tissues for RNA sequencing. Reverse transcription-quantitative PCR (RT-qPCR), Sanger sequencing, and fluorescence in situ hybridization were utilized to study the expression, localization, and clinical significance of circTRIO in LSCC tissues, and TU212 and TU686 cell lines. Furthermore, cell counting Kit-8, colony-forming assay, Transwell, and flow cytometry assays were assessed to illustrate the crucial role played by the circTRIO in proliferation, colony-forming ability, migration, and apoptosis in LSCC cells. Finally, the molecule's role as a microRNA (miRNA) sponge was analyzed. In the results, we screened out a promising upregulated novel circRNA-circTRIO in LSCC tumor tissues compared with paracancerous tissues using RNA sequencing. Then, we used qPCR to evaluate the expression of circTRIO in 20 additional paired LSCC tissues and two cells, and the results showed that circTRIO was highly expressed in LSCC and that this high expression was closely related to the malignant progression of LSCC. Furthermore, we examined circTRIO expression in the Gene Expression Omnibus data sets GSE142083 and GSE27020, and circTRIO expression was considerably higher in tumor tissues compared with the adjacent tissues. Kaplan-Meier survival analysis showed that the expression of circTRIO was associated with worse disease-free survival. The Gene Set Enrichment Analysis biological pathway evaluation results demonstrated that circTRIO was mainly enriched in cancer pathways. Moreover, we confirmed that silencing circTRIOs can help to significantly inhibit LSCC cell proliferation and migration while triggering apoptosis. Upregulated circTRIO expression levels may play an important role in the tumorigenesis and development of LSCC.
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Carcinoma de Células Escamosas , Neoplasias de Cabeça e Pescoço , Neoplasias Laríngeas , MicroRNAs , Humanos , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , RNA Circular/genética , Carcinoma de Células Escamosas/patologia , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/patologia , Hibridização in Situ Fluorescente , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias de Cabeça e Pescoço/genética , Carcinogênese/genética , Proliferação de Células/genética , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica/genéticaRESUMO
Circular RNAs (circRNAs) possess important functions in cervical carcinogenesis by operating as competing endogenous RNAs (ceRNAs). Our preliminary bioinformatics predicted the potential circ_0000212/microRNA (miR)-1236-3p/gremlin 1 (GREM1) ceRNA crosstalk. Thus, we further elucidated whether the novel ceRNA crosstalk can participate in cervical cancer development. Circ_0000212, miR-1236-3p and GREM1 were quantified by real-time quantitative polymerase chain reaction (qPCR) and immunoblotting. 5-ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry, and tube formation assay were performed to assess cell proliferation, apoptosis and tube formation, respectively. Transwell assay was used to detect cell migration and invasion. Mouse xenografts were established to evaluate the role of circ_0000212 in vivo. Dual-luciferase reporter assay was performed to verify the direct relationship between miR-1236-3p and circ_0000212 or GREM1. Circ_0000212 expression was elevated in human cervical cancer. Silencing of endogenous circ_0000212 hindered cancer cell proliferation, motility and invasion and induced apoptosis, as well as diminished the tube formation of human umbilical vein endothelial cells (HUVECs) in vitro. Circ_0000212 silencing also weakened tumor growth in vivo. Mechanistically, circ_0000212 directly bound to miR-1236-3p, and downregulation of miR-1236-3p reversed these effects of circ_0000212 silencing on cell malignant phenotypes and HUVEC tube formation. GREM1 was a direct miR-1236-3p target, and its expression was regulated by circ_0000212 through miR-1236-3p. Moreover, miR-1236-3p upregulation impeded cancer cell malignant phenotypes and HUVEC tube formation by targeting GREM1. Our findings identify a novel ceRNA regulatory network, circ_0000212/miR-1236-3p/GREM1 axis, in cervical carcinogenesis, and provide potential targets that can be explored for therapeutic interventions.
Assuntos
MicroRNAs , Neoplasias do Colo do Útero , Animais , Feminino , Humanos , Camundongos , Carcinogênese/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Células Endoteliais da Veia Umbilical Humana , Peptídeos e Proteínas de Sinalização Intercelular , MicroRNAs/genética , Neoplasias do Colo do Útero/genética , RNA Circular/genéticaRESUMO
TEAD3 acts as a transcription factor in many tumors to promote tumor occurrence and development. But in prostate cancer (PCa), it appears as a tumor suppressor gene. Recent studies have shown that this may be related to subcellular localization and posttranslational modification. We found that TEAD3 was down-expressed in PCa. Immunohistochemistry of clinical PCa specimens confirmed that TEAD3 expression was the highest in benign prostatic hyperplasia (BPH) tissues, followed by primary PCa tissues, and the lowest in metastatic PCa tissues, and its expression level was positively correlated with overall survival. MTT assay, clone formation assay, and scratch assay confirmed that overexpression of TEAD3 could significantly inhibit the proliferation and migration of PCa cells. Next-generation sequencing results indicated that Hedgehog (Hh) signaling pathway was significantly inhibited after overexpression of TEAD3. Rescue assays suggested that ADRBK2 could reverse the proliferation and migration ability caused by overexpression of TEAD3. TEAD3 is downregulated in PCa and associated with poor patient prognosis. Overexpression of TEAD3 inhibits the proliferation and migration ability of PCa cells via restraining the mRNA level of ADRBK2. These results indicate that TEAD3 was down-expressed in PCa patients and was positively correlated with a high Gleason score and poor prognosis. Mechanistically, we found that the upregulation of TEAD3 inhibits the proliferation and metastasis of prostate cancer by inhibiting the expression of ADRBK2.
Assuntos
Hiperplasia Prostática , Neoplasias da Próstata , Masculino , Humanos , Proteínas Hedgehog/metabolismo , Linhagem Celular Tumoral , Neoplasias da Próstata/patologia , Hiperplasia Prostática/genética , Hiperplasia Prostática/metabolismo , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Movimento Celular/genética , Fatores de Transcrição de Domínio TEARESUMO
Melanoma is the most aggressive cutaneous tumor. Neuropilin and tolloid-like 2 (NETO2) is closely related to tumorigenesis. However, the functional significance of NETO2 in melanoma progression remains unclear. Herein, we found that NETO2 expression was augmented in melanoma clinical tissues and associated with poor prognosis in melanoma patients. Disrupting NETO2 expression markedly inhibited melanoma proliferation, malignant growth, migration, and invasion by downregulating the levels of calcium ions (Ca2+) and the expression of key genes involved in the calcium signaling pathway. By contrast, NETO2 overexpression had the opposite effects. Importantly, pharmacological inhibition of CaMKII/CREB activity with the CaMKII inhibitor KN93 suppressed NETO2-induced proliferation and melanoma metastasis. Overall, this study uncovered the crucial role of NETO2-mediated regulation in melanoma progression, indicating that targeting NETO2 may effectively improve melanoma treatment.
Assuntos
Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina , Melanoma , Humanos , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Melanoma/genética , Proteínas de Membrana/genética , Fosforilação , Transdução de SinaisRESUMO
Permeable sandy sediments cover 50-60 % of the global continental shelf and are important bioreactors that regulate organic matter (OM) turnover and nutrient cycling in the coastal ocean. In sands, the dynamic porewater advection can cause rapid mass transfer and variable redox conditions, thus affecting OM remineralization pathways, as well as the recycling of iron and phosphorus. In this study, North Sea sands were incubated in flow-through reactors (FTRs) to investigate biogeochemical processes under porewater advection and changing redox conditions. We found that the average rate of anaerobic OM remineralization was 12 times lower than the aerobic pathway, and Fe(III) oxyhydroxides were found to be the major electron acceptors during 34 days of anoxic incubation. Reduced Fe accumulated in the solid phase (expressed as Fe(II)) before significant release of Fe2+ into the porewater, and most of the reduced Fe (~96 %) remained in the solid phase throughout the anoxic incubation. Fe(II) retained in the solid phase, either through the formation of authigenic Fe(II)-bearing minerals or adsorption, was easily re-oxidized upon exposure to O2. Excessive P release (apart from OM remineralization) started at the beginning of the anoxic incubation and accelerated after the release of Fe2+ with a constant P/Fe2+ ratio of 0.26. After 34 days of anoxic incubation, porewater was reoxygenated and > 99 % of released P was coprecipitated through Fe2+ oxidation (so-called "Fe curtain"). Our results demonstrate that Fe(III)/Fe(II) in the solid phase can serve as a relatively immobile and rechargeable "redox battery" under dynamic porewater advection. This Fe "redox battery" is characteristic for permeable sediments and environments with variable redox conditions, making Fe an important player in OM turnover. We also suggest that P liberated before Fe2+ release can escape the "Fe curtain" in surface sediments, thus potentially increasing net benthic P efflux from permeable sediments under variable redox conditions.
RESUMO
BACKGROUND: Non-invasive imaging technologies for assessing axillary lymph node (ALN) metastasis of breast cancer are needed in clinical practice. PURPOSE: To explore the clinical value of intravoxel incoherent motion (IVIM) and diffusional kurtosis imaging (DKI) for predicting ALN metastasis of breast cancer. MATERIAL AND METHODS: A total of 194 patients with pathologically confirmed breast cancer who underwent IVIM and DKI examination were reviewed retrospectively. The IVIM derived parameters of D, D*, and f and DKI-derived parameters of MD and MK were measured. The independent samples t-test was used to compare the parameters between the ALN metastasis and non-ALN metastasis groups. Receiver operating characteristic (ROC) curve analysis was also performed. RESULTS: The D and MD in the ALN metastasis group were significantly lower than those in the non-ALN metastasis group (P < 0.001, P < 0.001). The D*, f, and MK were higher in the ALN metastasis group than in the non-ALN metastasis group (P = 0.015, P = 0.014, and P = 0.001, respectively). The area under the ROC curve (AUC) of D (0.768) was highest. In addition, the diagnostic efficiency of both IVIM and DKI were higher than that of the conventional MRI (P = 0.002, P = 0.048). The diagnostic efficiency of IVIM + DKI were higher than that of the IVIM or DKI alone (P = 0.021, P = 0.004). CONCLUSION: IVIM and DKI can be used for predicting breast cancer ALN metastasis with D as the most meaningful parameter.