RESUMO
Ikaros transcription factors are essential for adaptive lymphocyte function, yet their role in innate lymphopoiesis is unknown. Using conditional genetic inactivation, we show that Ikzf1/Ikaros is essential for normal natural killer (NK) cell lymphopoiesis and IKZF1 directly represses Cish, a negative regulator of interleukin-15 receptor resulting in impaired interleukin-15 receptor signaling. Both Bcl2l11 and BIM levels, and intrinsic apoptosis were increased in Ikzf1-null NK cells, which in part accounts for NK lymphopenia as both were restored to normal levels when Ikzf1 and Bcl2l11 were co-deleted. Ikzf1-null NK cells presented extensive transcriptional alterations with reduced AP-1 transcriptional complex expression and increased expression of Ikzf2/Helios and Ikzf3/Aiolos. IKZF1 and IKZF3 directly bound AP-1 family members and deletion of both Ikzf1 and Ikzf3 in NK cells resulted in further reductions in Jun/Fos expression and complete loss of peripheral NK cells. Collectively, we show that Ikaros family members are important regulators of apoptosis, cytokine responsiveness and AP-1 transcriptional activity.
Assuntos
Células Matadoras Naturais , Fator de Transcrição AP-1 , Fator de Transcrição AP-1/genética , Células Matadoras Naturais/metabolismo , Receptores de Interleucina-15 , Fator de Transcrição Ikaros/genética , Fator de Transcrição Ikaros/metabolismoRESUMO
Although circumstantial evidence supports enhanced Toll-like receptor 7 (TLR7) signalling as a mechanism of human systemic autoimmune disease1-7, evidence of lupus-causing TLR7 gene variants is lacking. Here we describe human systemic lupus erythematosus caused by a TLR7 gain-of-function variant. TLR7 is a sensor of viral RNA8,9 and binds to guanosine10-12. We identified a de novo, previously undescribed missense TLR7Y264H variant in a child with severe lupus and additional variants in other patients with lupus. The TLR7Y264H variant selectively increased sensing of guanosine and 2',3'-cGMP10-12, and was sufficient to cause lupus when introduced into mice. We show that enhanced TLR7 signalling drives aberrant survival of B cell receptor (BCR)-activated B cells, and in a cell-intrinsic manner, accumulation of CD11c+ age-associated B cells and germinal centre B cells. Follicular and extrafollicular helper T cells were also increased but these phenotypes were cell-extrinsic. Deficiency of MyD88 (an adaptor protein downstream of TLR7) rescued autoimmunity, aberrant B cell survival, and all cellular and serological phenotypes. Despite prominent spontaneous germinal-centre formation in Tlr7Y264H mice, autoimmunity was not ameliorated by germinal-centre deficiency, suggesting an extrafollicular origin of pathogenic B cells. We establish the importance of TLR7 and guanosine-containing self-ligands for human lupus pathogenesis, which paves the way for therapeutic TLR7 or MyD88 inhibition.
Assuntos
Mutação com Ganho de Função , Lúpus Eritematoso Sistêmico , Receptor 7 Toll-Like , Animais , Autoimunidade/genética , Linfócitos B , GMP Cíclico/análogos & derivados , Guanosina , Humanos , Lúpus Eritematoso Sistêmico/genética , Camundongos , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/metabolismo , Receptor 7 Toll-Like/genética , Receptor 7 Toll-Like/metabolismoRESUMO
Human DEAD/H box RNA helicase DDX6 acts as an oncogene in several different types of cancer, where it participates in RNA processing. Nevertheless, the role of DDX6 in pancreatic cancer (PC), together with the underlying mechanism, has yet to be fully elucidated. In the present study, compared with adjacent tissues, the level of DDX6 was abnormally increased in human PC tissues, and this increased level of expression was associated with poor prognosis. Furthermore, the role of DDX6 in PC was investigated by overexpressing or silencing the DDX6 in the PC cell lines, SW1990 and PaTu-8988t. A xenograft model was established by injecting nude mice with either DDX6-overexpressing or DDX6-silenced SW1990 cells. DDX6 overexpression promoted the proliferation and cell cycle transition, inhibited the cell apoptosis of PC cells, and accelerated tumor formation, whereas DDX6 knockdown elicited the opposite effects. DDX6 exerted positive effects on PC. RNA immunoprecipitation assay showed that DDX6 bound to kinesin family member C1 (KIFC1) mRNA, which was further confirmed by RNA pull-down assay. These results suggested that DDX6 positively regulated the expression of KIFC1. KIFC1 overexpression enhanced the proliferative capability of PC cells with DDX6 knockdown and inhibited their apoptosis. By contrast, DDX6 overexpression reversed the inhibitory effect of KIFC1 silencing on tumor proliferation. Subsequently, the transcription factor Yin Yang 1 (YY1) was shown to negatively regulate DDX6 at both the mRNA and protein levels. Dual-luciferase reporter assay verified that YY1 targeted the promoter of DDX6 and inhibited its transcription. High expression levels of YY1 decreased the proliferation of PC cells and promoted cell apoptosis, although these effects were reversed by DDX6 overexpression. Taken together, YY1 may target the DDX6/KIFC1 axis, thereby negatively regulating its expression, leading to an inhibitory effect on pancreatic tumor.
Assuntos
RNA Helicases DEAD-box , MicroRNAs , Neoplasias Pancreáticas , Fator de Transcrição YY1 , Animais , Humanos , Camundongos , Linhagem Celular Tumoral , Proliferação de Células , RNA Helicases DEAD-box/genética , RNA Helicases DEAD-box/metabolismo , Regulação Neoplásica da Expressão Gênica , Camundongos Nus , MicroRNAs/genética , Neoplasias Pancreáticas/patologia , Proteínas Proto-Oncogênicas/metabolismo , RNA Mensageiro , Fator de Transcrição YY1/genética , Fator de Transcrição YY1/metabolismoRESUMO
BACKGROUND: Nanobodies, also known as VHH or single-domain antibodies, are unique antibody fragments derived solely from heavy chains. They offer advantages of small molecules and conventional antibodies, making them promising therapeutics. The paratope is the specific region on an antibody that binds to an antigen. Paratope prediction involves the identification and characterization of the antigen-binding site on an antibody. This process is crucial for understanding the specificity and affinity of antibody-antigen interactions. Various computational methods and experimental approaches have been developed to predict and analyze paratopes, contributing to advancements in antibody engineering, drug development, and immunotherapy. However, existing predictive models trained on traditional antibodies may not be suitable for nanobodies. Additionally, the limited availability of nanobody datasets poses challenges in constructing accurate models. METHODS: To address these challenges, we have developed a novel nanobody prediction model, named NanoBERTa-ASP (Antibody Specificity Prediction), which is specifically designed for predicting nanobody-antigen binding sites. The model adopts a training strategy more suitable for nanobodies, based on an advanced natural language processing (NLP) model called BERT (Bidirectional Encoder Representations from Transformers). To be more specific, the model utilizes a masked language modeling approach named RoBERTa (Robustly Optimized BERT Pretraining Approach) to learn the contextual information of the nanobody sequence and predict its binding site. RESULTS: NanoBERTa-ASP achieved exceptional performance in predicting nanobody binding sites, outperforming existing methods, indicating its proficiency in capturing sequence information specific to nanobodies and accurately identifying their binding sites. Furthermore, NanoBERTa-ASP provides insights into the interaction mechanisms between nanobodies and antigens, contributing to a better understanding of nanobodies and facilitating the design and development of nanobodies with therapeutic potential. CONCLUSION: NanoBERTa-ASP represents a significant advancement in nanobody paratope prediction. Its superior performance highlights the potential of deep learning approaches in nanobody research. By leveraging the increasing volume of nanobody data, NanoBERTa-ASP can further refine its predictions, enhance its performance, and contribute to the development of novel nanobody-based therapeutics. Github repository: https://github.com/WangLabforComputationalBiology/NanoBERTa-ASP.
Assuntos
Anticorpos de Domínio Único , Sítios de Ligação de Anticorpos , Anticorpos de Domínio Único/química , Anticorpos , Sítios de Ligação , Especificidade de AnticorposRESUMO
BACKGROUND: Helix-loop-helix transcription factor 4 (HTF4) as an anti-cancer target has been reported in many human cancers, but limited data exists regarding the effect of HTF4 in pancreatic cancer. In this study, we aimed to investigate the role of HTF4 in pancreatic cancer. METHODS: The expression levels of HTF4 in clinical pancreatic cancer samples were measured. HTF4 was knocked down or overexpressed in pancreatic cancer cells and was subsequently tested for bio-function using in vitro assays and in vivo. The regulation of HTF4 on GID2 was assessed via bioinformatic tools and dual-luciferase reporter assay. RESULTS: We found that HTF4 was highly expressed in pancreatic cancer tissues and correlated with poor patient prognosis. In addition, knocking down HTF4 expression inhibited cell proliferation, migration, and invasion, whereas HTF4 overexpression exerted the opposite effect. Moreover, HTF4 promoted tumor growth and metastasis in pancreatic cancer. Further, HTF4 bound to the GID2 promoter region and promoted transcriptional activation of GID2 in pancreatic cancer cells. GID2 knockdown suppressed HTF4-induced malignant behaviors of pancreatic cancer cells. CONCLUSIONS: Our findings suggest that the HTF4/GID2 axis accelerates the progression of pancreatic cancer, providing a potential therapeutic target and prognostic indicator for the treatment of pancreatic cancer patients.
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Dysregulation of deubiquitinase or ubiquitinase-mediated protein expression contributes to various diseases, including cancer. In the present study, we identified GID2, a subunit of the glucose-induced degradation-deficient (GID) complex that functions as an E3 ubiquitin ligase, as a potential key candidate gene in pancreatic cancer (PC) progression. The functional role and potential mechanism of GID2 in PC progression were investigated. Integrated bioinformatics analysis was performed to identify differentially expressed genes in PC based on the Gene Expression Profiling Interactive Analysis data sets. We found that GID2 was upregulated in PC tissues and that a high level of GID2 expression in clinical PC samples was positively associated with tumor stage and poor survival. Functional assays elucidated that GID2 expression promoted cell growth in vitro and accelerated tumor growth in vivo. GID2 knockdown effectively attenuated the malignant behaviors of PC cells and tumor formation. Furthermore, the protein network that interacted with the GID2 protein was constructed based on the GeneMANIA website. Cyclin-dependent kinase inhibitor 3 (CDKN3), a cell cycle regulator, was identified as a potential target of the GID2 protein. We revealed that GID2 positively regulated CDKN3 expression and inhibited CDKN3 ubiquitination. Furthermore, CDKN3 downregulation reversed the promoting effects of GID2 on PC progression. Therefore, the present study demonstrated that GID2 might regulate PC progression by maintaining the stability of the CDKN3 protein. These findings highlight the potential roles of the GID2/CDKN3 axis as a potential therapeutic target in PC.
Assuntos
Genes cdc , Neoplasias Pancreáticas , Humanos , Proliferação de Células/genética , Ciclo Celular , Neoplasias Pancreáticas/genética , Apoptose/genética , Linhagem Celular Tumoral , Regulação Neoplásica da Expressão Gênica , Proteínas Inibidoras de Quinase Dependente de Ciclina/genética , Fosfatases de Especificidade Dupla/genética , Neoplasias PancreáticasRESUMO
Electrocatalytic reactions based on electron transfer mediators provide a simple and effective route for the development of convenient and sensitive electrochemical assays. Here, we report a novel electrocatalytic assay for detection of EDTA-Fe(III), which is widely used as a supplement in iron-fortified foods to reduce prevalence of iron deficiency. Unlike conventional electrochemical methods to detect Fe(III) ion, signaling mechanism of our electrocatalytic assay relies on the previously unexplored thionine-mediated electrochemical reduction of EDTA-Fe(III). This electrocatalytic detection method is sensitive for EDTA-Fe(III) detection in the linear concentration range from 10 to 750 µM with a detection limit of 2.5 µM. It is also specific enough and applicable to detection of EDTA-Fe(III) in real soy sauce samples with satisfactory recovery. The one-step electrocatalytic reduction for signal generation enables the direct and sensitive electrochemical detection of EDTA-Fe(III). We believe that this electrocatalytic assay can serve as a general platform for quantification of EDTA-Fe(III) in many EDTA-Fe(III)-fortified foods. And because thionine is increasingly used as a signal reporter in electrochemical DNA/aptamer sensors, the engineered electrocatalytic reaction of thionine-mediated electrochemical reduction of EDTA-Fe(III) will also provide a simple signal amplification means for the development of highly sensitive electrochemical biosensors.
Assuntos
Compostos Férricos , Alimentos de Soja , Ácido Edético , FenotiazinasRESUMO
A highly fluorescent nitrogen and sulfur codoped carbon nanoparticles (N,S-CNP) sample was obtained by microwave-assisted pyrolysis of citric acid and L-cysteine. After being purified by dialysis, the complexity and chemical composition of N,S-CNP were evaluated by ultra-performance liquid chromatography coupled with mass spectrometry (UPLC-MS) as well as by UPLC coupled with ultraviolet (UV) absorption and fluorescence detection (UPLC-UV/FLD) methods. By using the high-resolution UPLC separation, the N,S-CNP were well fractionated into six fractions within 3.5 min. Based on high-accuracy MS and tandem (MS/MS) analyses, the N,S-CNP species were revealed to display various chemical formulas, including (C12H16N2O7S2) n , (C9H13NO8S) n , (C18H20N2O14S2) n , (C18H20N2O12S2) n , (C9H11NO5S) n , and (C9H11NO7S) n . More importantly, our study disclosed unambiguously for the first time that the N,S-CNP species exist as supramolecular clusters with their individual monomer units linked together through noncovalent bonding forces. By using UPLC-UV/FLD analysis, the spectral characteristics of each N,S-CNP species were revealed. Each individual CNP species possesses its unique absorption and PL properties with absorption bands that are redshifted, whereas its emission bands are blueshifted with its elution order. This work highlights the merit of UPLC-MS together with UPLC-UV/FLD to investigate the chemical composition and the spectral properties of each individual N,S-CNP species. It is anticipated that our proposed methodology will open up a new venue in optimizing experimental conditions for producing specific N,S-CNP species of desired composition. Graphical Abstract Carbon nanoparticles synthesized by microwave-assisted pyrolysis of citric acid and L-cysteine exist as supramolecular clusters with their individual monomer units linked together by noncovalent interactions.
RESUMO
2-Alkylcyclobutanones (2-ACBs) are uniquely formed when triglycerides-containing food products are exposed to ionizing radiation. Thus, 2-ACBs have been used as marker molecules to identify irradiated food. Most methods to determine 2-ACBs involve mass spectrometric detection after chromatographic separation. The spectrofluorometer is rarely used to determine 2-ACBs because these molecules do not fluoresce. In this study, we developed an ultra-performance liquid chromatography (UPLC) method to determine 2-ACBs. 2-ACBs were converted into fluorophores after reacting with 1-naphthalenyl hydrazine to facilitate their sensitive and selective detection using a fluorescence detector (FLD). Analysis of 2-ACBs using our developed UPLC-FLD method allows sensitive determination of 2-ACBs at a detection limit of 2 ng 2-ACBs per g of fat (30 pg/injection), which is significantly lower than that of existing analytical methods. After validation for trueness and precision, the method was applied to γ-irradiated chicken samples to determine their 2-ACB content. Comparative studies employing liquid chromatography-tandem mass spectrometric method revealed no systematic difference between the two methods, thereby demonstrating that the proposed UPLC-FLD method can be suitably used to determine 2-ACBs in irradiated foodstuffs. Graphical Abstract Determination of radiation-induced food-borne 2-dodecylcyclobutanone and 2-tetradecylcyclobutanone by combining 1-naphthalenyl hydrazine derivatization and ultra-performance liquid chromatography with fluorescence detection.
Assuntos
Butanonas/análise , Cromatografia Líquida/métodos , Hidrazinas/química , Espectrometria de Fluorescência/métodos , Animais , Galinhas , Raios gama , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Stromal cell-derived factor 1 (SDF-1) is a chemokine that is expressed in some cancer cells and is involved in tumor cell migration and metastasis. CXCR7, a novel receptor for SDF-1, has been identified recently. Research has demonstrated that SDF-1/CXCR7 interaction could play an important role in cancer progression. In this study, we aimed to investigate the expression of the SDF-1/CXCR7 ligand receptor system and the relationship between this expressions and clinicopathological characteristics in pancreatic adenocarcinoma. METHODS: Expressions of SDF-1 and CXCR7 in 64 cases of pancreatic adenocarcinoma tissue and 24 cases of normal pancreatic tissue were detected immunohistochemically. RESULTS: Expressions of SDF-1 and CXCR7 were negative in normal pancreatic tissues. Respectively, positive expression rates of SDF-1 and CXCR7 in pancreatic adenocarcinoma were 45.3% and 51.6%. The expression of SDF-1 correlated with histological grades; the expression rate in moderate to low differentiation was higher than in high differentiation (P<0.05). The expression of CXCR7 positively correlated with lymph node metastasis (P<0.05). A log-rank test showed that the expression of SDF-1+/CXCR7+ correlated with poor prognosis (P<0.05). CONCLUSIONS: The SDF-1/CXCR7 receptor ligand system may take part in invasive progression and metastasis of pancreatic adenocarcinoma, and might be useful as an index for evaluating invasiveness and prognosis.
Assuntos
Adenocarcinoma/metabolismo , Biomarcadores Tumorais/metabolismo , Quimiocina CXCL12/metabolismo , Neoplasias Pancreáticas/metabolismo , Receptores CXCR/metabolismo , Adenocarcinoma/mortalidade , Adenocarcinoma/secundário , Adulto , Idoso , Idoso de 80 Anos ou mais , Progressão da Doença , Feminino , Seguimentos , Humanos , Técnicas Imunoenzimáticas , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Neoplasias Pancreáticas/mortalidade , Neoplasias Pancreáticas/patologia , Prognóstico , Taxa de Sobrevida , Neoplasias PancreáticasRESUMO
Digital transformation, based on digital technologies, has triggered economic growth in many industries and brought about production and service transformation in the manufacturing sector. As an important source of innovation output and a driving force for national economic development, it is of great significance to study the impact of digital transformation on innovation output in manufacturing companies. This study analyzes the effects of digital transformation on the quality, quantity, and overall innovation output of manufacturing companies from both the macro provincial-level digital transformation and micro enterprise-level digital transformation perspectives. Additionally, using data from manufacturing companies listed on the Shanghai and Shenzhen stock exchanges from 2012 to 2022, this study empirically tests the mechanism through which digital transformation affects innovation output from the perspectives of internal transaction costs and external transaction costs. The results show that digital transformation promotes overall improvement in innovation output of manufacturing companies and leads to improvements in both the quality and quantity of innovation output. Furthermore, the study finds that the effect of digital transformation on innovation output has a nonlinear characteristic under different levels of market competitiveness and market freedom. The mediation analysis reveals that the influence of digital transformation on innovation output can be attributed to the reduction of internal transaction costs and the enhancement of external transaction efficiency. In terms of digital policy formulation, it is necessary to coordinate the development of diverse and innovative digital infrastructure at the macro level with the micro-level ecosystems of enterprises, in order to reduce transaction costs within and outside innovative entities. Ultimately, it is essential for the government to foster a conducive free market environment that enhances transaction efficiency and timely regulates the excessive competition resulting from oligopolistic monopolies, thus maximizing the potential of digital transformation in promoting innovation output.
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Comércio , Ecossistema , China , Indústrias , Tecnologia DigitalRESUMO
Inositol Polyphosphate-5-Phosphatase J (INPP5J), a 5-phosphatase, has been identified as a tumor suppressor in several types of cancer. However, its role in pancreatic cancer (PC) is unknown. We found that the INPP5J expression was markedly lower in PC tissues (n = 50) compared to paired adjacent non-tumor tissues, and the lower INPP5J expression was relevant to a worse prognosis of PC patients. We thus proposed that INPP5J might inhibit PC progression and conducted gain-of- and loss-of-function experiments to test our hypothesis. Our results showed that overexpression of INPP5J inhibited cell proliferation, invasion, migration, and xenografted tumor of PC cells. INPP5J silencing showed the opposite effect. Pellino E3 Ubiquitin Protein Ligase 1 (PELI1) is one of the ubiquitin ligases known to promote ubiquitination of its downstream targets. We found that PELI1 could interact with INPP5J and promote the ubiquitination and degradation of INPP5J. PELI1 overexpression enhanced malignant behaviors of PC cells. However, INPP5J overexpression restored the alterations caused by PELI1 overexpression. In conclusion, the results suggest that the decreased INPP5J expression, caused by PELI1 through ubiquitination, may promote PC progression. The PELI1-INPP5J axis represents a potential therapeutic targetable node for PC.
Assuntos
Movimento Celular , Proliferação de Células , Camundongos Nus , Neoplasias Pancreáticas , Monoéster Fosfórico Hidrolases , Ubiquitina-Proteína Ligases , Ubiquitinação , Animais , Feminino , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Linhagem Celular Tumoral , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Camundongos Endogâmicos BALB C , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/genética , Monoéster Fosfórico Hidrolases/metabolismo , Proteólise , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Regulação para CimaRESUMO
ABSTRACT: The 210 Pb burden in the skeleton is a measurement value suitable for the estimation of the cumulative exposure to radon, based on which the resultant risk of lung cancer can be derived. There have been a handful of studies that successfully measured 210 Pb activity in the bones of volunteers who had chronic exposure to high concentrations of radon occupationally or in their residences. However, the quantitative relationship between measured 210 Pb activity and radon exposure remains elusive. Herein, we investigate the origin of the skeletal burden by employing the biokinetic model recommended by the International Commission on Radiological Protection and modeling various routes of intake. First, the baseline 210 Pb burden for the general public regarding eating assorted foodstuffs and breathing normal air is obtained. It is found that this baseline burden ranges between 7.3 to 46.5 Bq for a 50-y-old (male) person, which characterizes a large variance due to the uncertainty of each route of intake. Next, we concentrate on radon exposure by referring to two experimental studies where the accounts of exposure and the measured 210 Pb burden for each volunteer are documented in detail. From comparing our prediction and measurements, it is found that exposure to higher concentration of radon is the most significant source of 210 Pb intake, and the quantitative differences can be reasonably explained by the uncertainty resulting from regular intake routes. This study establishes the theoretical foundation for assessing one's risk of lung cancer due to radon exposure by measuring the 210 Pb burden in bones.
Assuntos
Osso e Ossos , Chumbo , Radônio , Radônio/análise , Humanos , Chumbo/análise , Masculino , Osso e Ossos/efeitos da radiação , Osso e Ossos/química , Osso e Ossos/metabolismo , Pessoa de Meia-Idade , Exposição à Radiação/análise , Feminino , Carga Corporal (Radioterapia) , Poluentes Radioativos do Ar/análise , Exposição Ambiental/análiseRESUMO
The counting efficiency calibration for in vivo measurement is crucial to derive the activity of radionuclides residing inside a monitored subject. Recently, virtual calibration based on computational phantoms has become popular, yet some key questions remain unresolved. Here, we focus on the in vivo measurement of Pb-210 in the skull and systematically examine how virtual calibration compares to those using physical phantoms and how the variety of computational phantoms affects the derived counting efficiency. It is found that the virtually calibrated efficiency based on the MIDA phantom, which characterizes the highest anatomical fidelity, shows reasonable consistency with the experimental counterpart, with a relative bias of approximately 10%. However, in comparison to the case based on the MIDA phantom, those based on the BOMAB and MIRD phantoms show larger deviation, demonstrating underestimations on the counting efficiency by 51% and 42%, respectively. This finding underscores the critical role of computational phantoms in the virtual calibration. This study contributes to the development of techniques for assessing lung cancer risk resulting from chronic radon exposure through in vivo measurement of skeletal Pb-210 activity.
Assuntos
Chumbo , Contagem Corporal Total , Contagem Corporal Total/métodos , Simulação por Computador , Calibragem , Método de Monte Carlo , Radioisótopos de Chumbo , Crânio , Imagens de FantasmasRESUMO
BACKGROUND: Lung cancer is a serious threat to human health and is the first leading cause of cancer death. Ferroptosis, a newly discovered form of programmed cell death associated with redox homeostasis, is of particular interest in the lung cancer, given the high oxygen environment of lung cancer. NADPH has reducing properties and therefore holds the potential to resist ferroptosis. Resistance to ferroptosis exists in lung cancer, but the role of NADK in regulating ferroptosis in lung cancer has not been reported yet. METHODS: Immunohistochemistry (IHC) was used to analyse the expression of NADK in 86 cases of lung adenocarcinoma(LUAD) and adjacent tissues, and a IHC score was assigned to each sample. Chi-square and kaplan-meier curve was performed to analyse the differences in metastasis and five-year survival between the two groups with NADK high or low scores. Proliferation of NADK-knockdown LUAD cell lines was detected in vivo and vitro. Furthermore, leves of ROS, MDA and Fe2+ were measured to validate the effect and mechanism of NADK on ferroptosis in LUAD. RESULTS: The expression of NADK was significantly evaluated in LUAD tissues as compared to adjacent non-cancerous tissues. The proliferation of NADK-knockdown cells was inhibited both in vivo and vitro, and increasing levels of intracellular ROS, Fe2+ and lipid peroxide products (MDA) were observed. Furthermore, NADK-knockdown promoted the ferroptosis of LUAD cells induced by Erastin/RSL3 by regulating the level of NADPH and the expression of FSP1. Knockdown of NADK enhanced the sensitivities of LUAD cells to Erastin/RSL3-induced ferroptosis by regulating NADPH level and FSP1 expression. CONCLUSIONS: NADK is over-expressed in LUAD patients. Knockdown of NADK inhibited the proliferation of LUAD cells both in vitro and in vivo and promotes the Erastin/RSL3-induced ferroptosis of LUAD cells by down-regulating the NADPH/FSP1 axis.
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Adenocarcinoma de Pulmão , Ferroptose , Neoplasias Pulmonares , NADP , Animais , Feminino , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Adenocarcinoma de Pulmão/patologia , Adenocarcinoma de Pulmão/genética , Adenocarcinoma de Pulmão/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Ferroptose/genética , Ferroptose/fisiologia , Técnicas de Silenciamento de Genes , Neoplasias Pulmonares/patologia , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Camundongos Nus , NADP/metabolismoRESUMO
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease with a clear genetic component. While most SLE patients carry rare gene variants in lupus risk genes, little is known about their contribution to disease pathogenesis. Amongst them, SH2B3-a negative regulator of cytokine and growth factor receptor signaling-harbors rare coding variants in over 5% of SLE patients. Here, we show that unlike the variant found exclusively in healthy controls, SH2B3 rare variants found in lupus patients are predominantly hypomorphic alleles, failing to suppress IFNGR signaling via JAK2-STAT1. The generation of two mouse lines carrying patients' variants revealed that SH2B3 is important in limiting the number of immature and transitional B cells. Furthermore, hypomorphic SH2B3 was shown to impair the negative selection of immature/transitional self-reactive B cells and accelerate autoimmunity in sensitized mice, at least in part due to increased IL-4R signaling and BAFF-R expression. This work identifies a previously unappreciated role for SH2B3 in human B cell tolerance and lupus risk.
Assuntos
Autoimunidade , Lúpus Eritematoso Sistêmico , Animais , Humanos , Camundongos , Autoimunidade/genética , Fator Ativador de Células B/metabolismo , Linfócitos B , Lúpus Eritematoso Sistêmico/genética , Células Precursoras de Linfócitos BRESUMO
Pancreatic cancer is one of the most aggressive cancers. PELI1 has been reported to promote cell survival and proliferation in multiple cancers. As of now, the role of PELI1 in pancreatic cancer is largely unknown. Here, we found that the PELI1 mRNA was higher expressed in pancreatic tumor tissues than in adjacent normal tissues, and the high PELI1 level in pancreatic cancer patients had a short survival time compared with the low level. Moreover, the results showed that PELI1 promoted cell proliferation, migration, and invasion, and inhibited apoptosis in vitro. Xenograft tumor experiments were used to determine the biological function of PELI1, and the results showed that PELI1 promoted tumor growth in vivo. Additionally, we found that Jagged1 activated PELI1 transcription in pancreatic cancer cells. To sum up, our results show that PELI1 affects the malignant phenotype of pancreatic cancer.
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Neoplasias Pancreáticas , Humanos , Apoptose/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Ubiquitina-Proteína Ligases/metabolismo , Neoplasias PancreáticasRESUMO
Cu-9Ni-6Sn alloys have received widespread attention due to their good mechanical properties and resistance to stress relaxation in the electronic and electrical industries. The hot compression deformation behaviors of the Cu-9Ni-6Sn-0.3Mn-0.2Zn alloy were investigated using the Gleeble-3500 thermal simulator at a temperature range of 700-900 °C and a strain rate range of 0.001-1 s-1. The microstructural evolution of the Cu-9Ni-6Sn alloy during hot compression was studied by means of an optical microscope and a scanning electron microscope. The constitutive equation of hot compression of the alloy was constructed by peak flow stress, and the corresponding 3D hot processing maps were plotted. The results showed that the peak flow stress decreased with the increase in the compression temperature and the decrease in the strain rate. The hot deformation activation energy was calculated as 243.67 kJ/mol by the Arrhenius equation, and the optimum deformation parameters for the alloy were 740-760 °C and 840-900 °C with a strain rate of 0.001~0.01 s-1. According to Deform-3D finite element simulation results, the distribution of the equivalent strain field in the hot deformation samples was inhomogeneous. The alloy was more sensitive to the deformation rate than to the temperature. The simulation results can provide a guideline for the optimization of the microstructure and hot deformation parameters of the Cu-9Ni-6Sn-0.3Mn-0.2Zn alloy.
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Proteasome 26S subunit, non-ATPase 12 (PSMD12) genes have been implicated in several types of malignancies but the role of PSMD12 in pancreatic cancer (PC) remains elusive. Bioinformatics analysis showed that PSMD12 was highly expressed in PC patients and was associated with shorter overall survival. PSMD12 was also shown to be highly expressed in PC tissues and cell lines. Upregulated PSMD12 showed enhanced cell viability, increased colony formation rate and upregulated levels of PCNA and c-Myc, while the inhibition of PSMD12 abated these levels. PSMD12 knockdown promoted cell apoptosis. The results of xenografts in nude mice confirmed that PSMD12 promoted PC tumor growth in vivo. Proteinâprotein interaction network and functional enrichment analyses implied that PSMD12 may have a connection with cyclin-dependent kinase inhibitor 3 (CDKN3). Coimmunoprecipitation and western blot results confirmed that PSMD12 could interact with and abate the ubiquitination level of CDKN3, thus stabilizing the CDKN3 protein. Rescue assays showed that PSMD12 overexpression caused cell proliferation and that knockdown-induced cell apoptosis could be reversed by CDKN3 regulation. This work reveals the essential roles of PSMD12 in the proliferation and apoptosis of PC development. PSMD12 may regulate CDKN3 expression by interacting with and abating the ubiquitination level of CDKN3, thereby participating in the malignant behavior of PC.