RESUMEN
Acute respiratory distress syndrome (ARDS) is a severe form of acute respiratory failure characterised by extensive inflammatory injury to the alveolocapillary barrier leading to alveolar oedema, impaired gas exchange and, ultimately, hypoxaemia necessitating the use of supplemental oxygen combined with some degree of positive airway pressure. Although much heterogeneity exists regarding the aetiology, localisation and endotypic characterisation of ARDS, what remains largely undisputed is the role of the innate immune system, and in particular of neutrophils, in precipitating and propagating lung injury. Activated neutrophils, recruited to the lung through chemokine gradients, promote injury by releasing oxidants, proteases and neutrophil extracellular traps, which ultimately cause platelet aggregation, microvascular thrombosis and cellular death. Among various neutrophilic chemoattractants, interleukin-8/C-X-C motif ligand 8 and related chemokines, collectively called ELR+ chemokines, acting on neutrophils through the G protein-coupled receptors CXCR1 and CXCR2, are pivotal in orchestrating the neutrophil activation status and chemotaxis in the inflamed lung. This allows efficient elimination of infectious agents while at the same time minimising collateral damage to host tissue. Therefore, understanding how CXCR1 and CXCR2 receptors are regulated is important if we hope to effectively target them for therapeutic use in ARDS. In the following narrative review, we provide an overview of the role of ELR+ chemokines in acute lung injury (ALI) and ARDS, we summarise the relevant regulatory pathways of their cognisant receptors CXCR1/2 and highlight current preclinical and clinical evidence on the therapeutic role of CXCR1 and CXCR2 inhibition in animal models of ALI, as well as in ARDS patients.
Asunto(s)
Pulmón , Neutrófilos , Receptores de Interleucina-8A , Receptores de Interleucina-8B , Síndrome de Dificultad Respiratoria , Transducción de Señal , Humanos , Síndrome de Dificultad Respiratoria/inmunología , Síndrome de Dificultad Respiratoria/metabolismo , Síndrome de Dificultad Respiratoria/terapia , Receptores de Interleucina-8B/metabolismo , Animales , Receptores de Interleucina-8A/metabolismo , Neutrófilos/metabolismo , Neutrófilos/inmunología , Pulmón/inmunología , Pulmón/fisiopatología , Pulmón/metabolismo , Activación Neutrófila , Infiltración NeutrófilaRESUMEN
Studies conducted on animal models have identified several therapeutic targets for myelofibrosis, the most severe of the myeloproliferative neoplasms. Unfortunately, many of the drugs which were effective in pre-clinical settings had modest efficacy when tested in the clinic. This discrepancy suggests that treatment for this disease requires combination therapies. To rationalize possible combinations, the efficacy in the Gata1low model of drugs currently used for these patients (the JAK1/2 inhibitor Ruxolitinib) was compared with that of drugs targeting other abnormalities, such as p27kip1 (Aplidin), TGF-ß (SB431542, inhibiting ALK5 downstream to transforming growth factor beta (TGF-ß) signaling and TGF-ß trap AVID200), P-selectin (RB40.34), and CXCL1 (Reparixin, inhibiting the CXCL1 receptors CXCR1/2). The comparison was carried out by expressing the endpoints, which had either already been published or had been retrospectively obtained for this study, as the fold change of the values in the corresponding vehicles. In this model, only Ruxolitinib was found to decrease spleen size, only Aplidin and SB431542/AVID200 increased platelet counts, and with the exception of AVID200, all the inhibitors reduced fibrosis and microvessel density. The greatest effects were exerted by Reparixin, which also reduced TGF-ß content. None of the drugs reduced osteopetrosis. These results suggest that future therapies for myelofibrosis should consider combining JAK1/2 inhibitors with drugs targeting hematopoietic stem cells (p27Kip1) or the pro-inflammatory milieu (TGF-ß or CXCL1).
Asunto(s)
Janus Quinasa 1 , Selectina-P , Mielofibrosis Primaria , Pirimidinas , Receptores de Interleucina-8B , Factor de Crecimiento Transformador beta , Mielofibrosis Primaria/tratamiento farmacológico , Mielofibrosis Primaria/metabolismo , Mielofibrosis Primaria/patología , Factor de Crecimiento Transformador beta/metabolismo , Animales , Janus Quinasa 1/antagonistas & inhibidores , Janus Quinasa 1/metabolismo , Selectina-P/metabolismo , Receptores de Interleucina-8B/antagonistas & inhibidores , Receptores de Interleucina-8B/metabolismo , Pirimidinas/farmacología , Pirimidinas/uso terapéutico , Receptores de Interleucina-8A/antagonistas & inhibidores , Receptores de Interleucina-8A/metabolismo , Ratones , Janus Quinasa 2/metabolismo , Janus Quinasa 2/antagonistas & inhibidores , Nitrilos/uso terapéutico , Nitrilos/farmacología , Modelos Animales de Enfermedad , Quimioterapia Combinada , Factor de Transcripción GATA1/metabolismo , Factor de Transcripción GATA1/genética , Pirazoles/farmacología , Pirazoles/uso terapéutico , HumanosRESUMEN
The level of tumor and circulating CXCR1/2-expressing neutrophils and CXCR1/2 ligands correlate with poor patient outcomes, inversely correlate with tumoral lymphocyte content, and predict immune checkpoint inhibitor (ICI) treatment failure. Accordingly, CXCR2-selective and CXCR1/2 dual inhibitors exhibit activity both as single agents and in combination with ICI treatment in mouse tumor models. Based on such reports, clinical trials combining CXCR1/2 axis antagonists with ICI treatment for cancer patients are underway. It has been assumed that CXCR1/2 blockade impacts tumors by blocking neutrophil chemotaxis and reducing neutrophil content in tumors. Here, we show that while CXCR2 antagonism does slow tumor growth, it does not preclude neutrophil recruitment into tumor. Instead, CXCR1/2 inhibition alters neutrophil function by blocking the polarization of transcriptional programs toward immune suppressive phenotypes and rendering neutrophils incapable of suppressing lymphocyte proliferation. This is associated with decreased release of reactive oxygen species and Arginase-1 into the extracellular milieu. Remarkably, these therapeutics do not impact the ability of neutrophils to phagocytose and kill ingested bacteria. Taken together, these results mechanistically explain why CXCR1/2 inhibition has been active in cancer but without infectious complications.
Asunto(s)
Neutrófilos , Receptores de Interleucina-8A , Receptores de Interleucina-8B , Receptores de Interleucina-8B/antagonistas & inhibidores , Receptores de Interleucina-8B/metabolismo , Receptores de Interleucina-8A/antagonistas & inhibidores , Receptores de Interleucina-8A/metabolismo , Neutrófilos/efectos de los fármacos , Neutrófilos/inmunología , Neutrófilos/metabolismo , Animales , Ratones , Humanos , Infiltración Neutrófila/efectos de los fármacos , Neoplasias/tratamiento farmacológico , Neoplasias/inmunología , Neoplasias/patología , Neoplasias/metabolismo , Inhibidores de Puntos de Control Inmunológico/farmacología , Inhibidores de Puntos de Control Inmunológico/uso terapéutico , Línea Celular Tumoral , Ratones Endogámicos C57BL , FemeninoRESUMEN
Pulmonary adenocarcinoma (PADC) treatment limited efficacy in preventing tumor progression, often resulting in malignant pleural effusion (MPE). MPE is filled with various mediators, especially interleukin-8 (IL-8). However, the role of IL-8 and its signaling mechanism within the fluid microenvironment (FME) implicated in tumor progression warrants further investigation. Primary cultured cells from samples of patients with MPE from PADC, along with a commonly utilized lung cancer cell line, were employed to examine the role of IL-8 and its receptor, CXCR1, through comparative analysis. Our study primarily assessed migration and invasion capabilities, epithelial-mesenchymal transition (EMT), and cancer stem cell (CSC) properties. Additionally, IL-8 levels in MPE fluid versus serum, along with immunohistochemical expression of IL-8/CXCR1 signaling in tumor tissue and cell blocks were analyzed. IL-8/CXCR1 overexpression enhanced EMT and CSC properties. Furthermore, the immunocytochemical examination of 17 cell blocks from patients with PADC and MPE corroborated the significant correlation between upregulated IL-8 and CXCR1 expression and the co-expression of IL-8 and CXCR1 in MPE with distant metastasis. In summary, the IL-8/ CXCR1 axis in FME is pivotal to tumor promotion via paracrine and autocrine signaling. Our study provides a therapeutic avenue for improving the prognosis of PADC patients with MPE.
Asunto(s)
Adenocarcinoma del Pulmón , Transición Epitelial-Mesenquimal , Interleucina-8 , Neoplasias Pulmonares , Derrame Pleural Maligno , Receptores de Interleucina-8A , Transducción de Señal , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adenocarcinoma del Pulmón/patología , Adenocarcinoma del Pulmón/metabolismo , Adenocarcinoma del Pulmón/genética , Adenocarcinoma del Pulmón/complicaciones , Línea Celular Tumoral , Movimiento Celular , Progresión de la Enfermedad , Interleucina-8/metabolismo , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/genética , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Derrame Pleural Maligno/patología , Derrame Pleural Maligno/metabolismo , Receptores de Interleucina-8A/metabolismo , Receptores de Interleucina-8A/genética , Microambiente TumoralRESUMEN
BACKGROUND: The prevalence of type 2 diabetic nephropathy (T2DN) ranges from 20 % to 40 % among individuals with type 2 diabetes. Multiple immune pathways play a pivotal role in the pathogenesis of T2DN. This study aimed to investigate the immunomodulatory effects of active ingredients derived from 14 traditional Chinese medicines (TCMs) on T2DN. METHODS: By removing batch effect on the GSE30528 and GSE96804 datasets, we employed a combination of weighted gene co-expression network analysis, least absolute shrinkage and selection operator analysis, protein-protein interaction network analysis, and the CIBERSORT algorithm to identify the active ingredients of TCMs as well as potential hub biomarkers associated with immune cells. Functional analysis was conducted using Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and gene set variation analysis (GSVA). Additionally, molecular docking was employed to evaluate interactions between active ingredients and potential immunotherapy targets. RESULTS: A total of 638 differentially expressed genes (DEGs) were identified in this study, comprising 5 hub genes along with 4 potential biomarkers. Notably, CXCR1, CXCR2, and FOS exhibit significant associations with immune cells while displaying robust or favorable affinities towards the active ingredients kaempferol, quercetin, and luteolin. Furthermore, functional analysis unveiled intricate involvement of DEGs, hub genes and potential biomarkers in pathways closely linked to immunity and diabetes. CONCLUSION: The potential hub biomarkers and immunotherapy targets associated with immune cells of T2DN comprise CXCR1, CXCR2, and FOS. Furthermore, kaempferol, quercetin, and luteolin demonstrate potential immunomodulatory effects in modulating T2DN through the regulation of CXCR1, CXCR2, and FOS expression.
Asunto(s)
Biología Computacional , Diabetes Mellitus Tipo 2 , Nefropatías Diabéticas , Medicamentos Herbarios Chinos , Medicina Tradicional China , Simulación del Acoplamiento Molecular , Farmacología en Red , Mapas de Interacción de Proteínas , Receptores de Interleucina-8B , Nefropatías Diabéticas/tratamiento farmacológico , Nefropatías Diabéticas/genética , Nefropatías Diabéticas/inmunología , Humanos , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/inmunología , Diabetes Mellitus Tipo 2/genética , Medicamentos Herbarios Chinos/uso terapéutico , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , Receptores de Interleucina-8B/genética , Receptores de Interleucina-8B/metabolismo , Receptores de Interleucina-8A/genética , Receptores de Interleucina-8A/metabolismo , Redes Reguladoras de Genes/efectos de los fármacosRESUMEN
BCR::ABL1 negative myeloproliferative neoplasms (MPN) form a distinct group of hematologic malignancies characterized by sustained proliferation of cells from multiple myeloid lineages. With a median survival of 16-35 months in patients with high-risk disease, primary myelofibrosis (PMF) is considered the most aggressive entity amongst all BCR::ABL1 MPN. Additionally, for a significant subset of patients, MPN evolve into secondary acute myeloid leukemia (AML), which has an even poorer prognosis compared to de novo AML. As the exact mechanisms of disease development and progression remain to be elucidated, current therapeutic approaches fail to prevent disease progression or transformation into secondary AML. As each MPN entity is characterized by sustained activation of various immune cells and raised cytokine concentrations within bone marrow (BM) and peripheral blood (PB), MPN may be considered to be typical inflammation-related malignancies. However, the exact role and consequences of increased cytokine concentrations within BM and PB plasma has still not been completely established. Up-regulated cytokines can stimulate cellular proliferation, or contribute to the development of an inflammation-related BM niche resulting in genotoxicity and thereby supporting mutagenesis. The neutrophil chemoattractant CXCL8 is of specific interest as its concentration is increased within PB and BM plasma of patients with PMF. Increased concentration of CXCL8 negatively correlates with overall survival. Furthermore, blockage of the CXCR1/2 axis appears to be able to reduce BM fibrosis and megakaryocyte dysmorphia in murine models. In this review, we summarize available evidence on the role of the CXCL8-CXCR1/2 axis within the pathogenesis of PMF, and discuss potential therapeutic modalities targeting either CXCL8 or its cognate receptors CXCR1/2.
Asunto(s)
Interleucina-8 , Mielofibrosis Primaria , Receptores de Interleucina-8A , Receptores de Interleucina-8B , Animales , Humanos , Interleucina-8/metabolismo , Mielofibrosis Primaria/metabolismo , Mielofibrosis Primaria/patología , Mielofibrosis Primaria/genética , Mielofibrosis Primaria/diagnóstico , Mielofibrosis Primaria/etiología , Mielofibrosis Primaria/mortalidad , Receptores de Interleucina-8A/metabolismo , Receptores de Interleucina-8A/genética , Receptores de Interleucina-8B/metabolismo , Receptores de Interleucina-8B/genética , Transducción de SeñalRESUMEN
OBJECTIVE@#To investigate and analyze the effect of CXC chemokine receptor 1/2 (CXCR1/2) targeting inhibitor Reparixin combined with cytarabine (Ara-C) on the malignant biological behaviors of acute myeloid leukemia cells and its effect on the expression of the CXCR family, while exploring the accompanying molecular mechanism, providing scientific basis and reference for new molecular markers and targeted therapy for AML.@*METHODS@#Acute myeloid leukemia U937 cells were treated with different concentrations of Reparixin, Ara-C alone or in combination, and the cell morphology was observed under an inverted microscope; Wright-Giemsa staining was used to detect cell morphological changes; CCK-8 method was used to detect cell proliferation; the ability of cell invasion was detected by Transwell chamber method; the ability of colony formation was detected by colony formation assay; cell apoptosis was detected by Hoechst 33258 fluorescent staining and Annexin V/PI double-staining flow cytometry; monodansylcadaverine(MDC) staining was used to detect cell autophagy; the expression of apoptosis, autophagy and related signaling pathway proteins was detected by Western blot and the expression changes of CXCR family were detected by real-time quantitative polymerase chain reaction (qRT-PCR).@*RESULTS@#Reparixin could inhibit the proliferation, invasion, migration and clone formation ability of U937 cells. Compared with the single drug group, when U937 cells were intervened by Reparixin combined with Ara-C, the malignant biological behaviors such as proliferation, invasion and colony formation were significantly decreased, and the levels of apoptosis and autophagy were significantly increased (P<0.01). After Reparixin combined with Ara-C intervenes in U937 cells, it can up-regulate the expression of the pro-apoptotic protein Bax and significantly down-regulate the expression of the anti-apoptotic protein Bcl-2, and also hydrolyze and activate Caspase-3, thereby inducing cell apoptosis. Reparixin combined with Ara-C could up-regulate the expressions of LC3Ⅱ and Beclin-1 proteins in U937 cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (P<0.01). MDC result showed that the green granules of vesicles increased significantly, and a large number of broken cells were seen (P<0.01). Reparixin combined with Ara-C can significantly inhibit the phosphorylation level of PI3K, AKT and NF-κB signaling molecule, inhibit the malignant biological behavior of cells by inhibiting the activation of PI3K/AKT/NF-κB pathway, and induce programmed cell death. Ara-C intervention in U937 cells had no effect on the expression of CXCR family (P>0.05). The expression of CXCR1, CXCR2, and CXCR4 mRNA could be down-regulated by Reparixin single-agent intervention in U937 cells (P<0.05), and the expression of CXCR2 was more significantly down-regulated than the control group and other CXCRs (P<0.01). When Reparixin and Ara-C intervened in combination, the down-regulated levels of CXCR1 and CXCR2 were more significant than those in the single-drug group (P<0.01), while the relative expressions of CXCR4 and CXCR7 mRNA had no significant difference compared with the single-drug group (P>0.05).@*CONCLUSION@#Reparixin combined with Ara-C can synergistically inhibit the malignant biological behaviors of U937 cells such as proliferation, invasion, migration and clone formation, and induce autophagy and apoptosis. The mechanism may be related to affecting the proteins expression of Bcl-2 family and down-regulating the proteins expression of CXCR family, while inhibiting the PI3K/AKT/NF-κB signaling pathway.
Asunto(s)
Humanos , Células U937 , Citarabina/uso terapéutico , Receptores de Interleucina-8A , FN-kappa B , Proteínas Proto-Oncogénicas c-akt , Fosfatidilinositol 3-Quinasas , Leucemia Mieloide Aguda/genética , Apoptosis , Proliferación Celular , Proteínas Reguladoras de la Apoptosis , Proteínas Proto-Oncogénicas c-bcl-2 , ARN Mensajero , Línea Celular TumoralRESUMEN
Abstract The current study aimed to assess whether the A122V causal polymorphism promotes alterations in the functional and structural proprieties of the CXC chemokine receptor type 1 protein (CXCR1) of cattle Bos taurus by in silico analyses. Two amino acid sequences of bovine CXCR1 was selected from database UniProtKB/Swiss-Prot: a) non-polymorphic sequence (A7KWG0) with alanine (A) at position 122, and b) polymorphic sequence harboring the A122V polymorphism, substituting alanine by valine (V) at same position. CXCR1 sequences were submitted as input to different Bioinformatics' tools to examine the effects of this polymorphism on functional and structural stabilities, to predict eventual alterations in the 3-D structural modeling, and to estimate the quality and accuracy of the predictive models. The A122V polymorphism exerted tolerable and non-deleterious effects on the polymorphic CXCR1, and the predictive structural model for polymorphic CXCR1 revealed an alpha helix spatial structure typical of a receptor transmembrane polypeptide. Although higher variations in the distances between pairs of amino acid residues at target-positions are detected in the polymorphic CXCR1 protein, more than 97% of the amino acid residues in both models were located in favored and allowed conformational regions in Ramachandran plots. Evidences has supported that the A122V polymorphism in the CXCR1 protein is associated with increased clinical mastitis incidence in dairy cows. Thus, the findings described herein prove that the replacement of the alanine by valine amino acids provokes local conformational changes in the A122V-harboring CXCR1 protein, which could directly affect its post-translational folding mechanisms and biological functionality.
Resumo O presente estudo objetivou avaliar se o polimorfismo causal A122V promove alterações nas propriedades funcionais e estruturais da proteína receptora de quimiocina CXC do tipo 1 (CXCR1) de bovino Bos taurus por análises in silico. Duas sequências de aminoácidos da CXCR1 bovina foram selecionadas a partir do banco de dados UniProtKB/Swiss-Prot: a) sequência não-polimórfica (A7KWG0) contendo alanina (A) na posição 122, e b) sequência polimórfica carreando o polimorfismo A122V, causando a substituição de alanina por valina (V) na mesma posição. As sequências CXCR1 foram analisadas por diferentes ferramentas de Bioinformática para examinar o efeito desse polimorfismo sobre sua estabilidade, função e estrutura, predizer eventuais alterações na sua modelagem estrutural 3-D, bem como estimar a qualidade dos modelos preditos. O polimorfismo A122V exerceu efeitos toleráveis e não-deletérios sobre a CXCR1 polimórfica, apresentando um modelo estrutural de alfa-hélice típico de uma proteína receptora transmembranar para ambas as proteínas. Embora maiores variações nas distâncias entre os pares de aminoácidos nas posições-alvo tenham sido detectadas na proteína polimórfica, mais do que 97% dos aminoácidos em ambos os modelos foram situados em regiões ditas favoráveis e permitidas nos diagramas de Ramachandran. Evidências sustentam que o polimorfismo de nucleotídeo único A122V na proteína receptora CXCR1 está associado à aumentada incidência de mastite clínica em vacas leiteiras. Assim, as descobertas descritas aqui comprovam que a substituição do aminoácido alanina por valina provoca mudanças conformacionais locais na proteína CXCR1 polimórfica, que podem estar diretamente afetando seus mecanismos de enovelamento pós-traducionais e sua função biológica.
Asunto(s)
Animales , Femenino , Polimorfismo de Nucleótido Simple , Receptores de Interleucina-8A , Bovinos , Secuencia de AminoácidosRESUMEN
Abstract Brain tumors are one of the most common causes of cancer-related deaths around the world. Angiogenesis is critical in high-grade malignant gliomas, such as glioblastoma multiforme. Objective: The aim of this study is to comparatively analyze the angiogenesis-related genes, namely VEGFA, VEGFB, KDR, CXCL8, CXCR1 and CXCR2 in LGG vs. GBM to identify molecular distinctions using datasets available on The Cancer Genome Atlas (TCGA). Methods: DNA sequencing and mRNA expression data for 514 brain lower grade glioma (LGG) and 592 glioblastoma multiforme (GBM) patients were acquired from The Cancer Genome Atlas (TCGA), and the genetic alterations and expression levels of the selected genes were analyzed. Results: We identified six distinct KDR mutations in the LGG patients and 18 distinct KDR mutations in the GBM patients, including missense and nonsense mutations, frame shift deletion and altered splice region. Furthermore, VEGFA and CXCL8 were significantly overexpressed within GBM patients. Conclusions: VEGFA and CXCL8 are important factors for angiogenesis, which are suggested to have significant roles during tumorigenesis. Our results provide further evidence that VEGFA and CXCL8 could induce angiogenesis and promote LGG to progress into GBM. These findings could be useful in developing novel targeted therapeutics approaches in the future.
Resumo Os tumores cerebrais são uma das causas mais comuns de mortes relacionadas ao câncer em todo o mundo. A angiogênese tem caráter crítico em gliomas malignos de alto grau, como o glioblastoma multiforme. Objetivo: O objetivo deste estudo foi analisar comparativamente os genes relacionados à angiogênese, VEGFA, VEGFB, KDR, CXCL8, CXCR1 e CXCR2 em GBG vs. GBM para identificar distinções moleculares usando conjuntos de dados disponíveis no The Cancer Genome Atlas (TCGA). Métodos: Os dados de sequenciamento de DNA e expressão de mRNA para 514 pacientes com glioma cerebral de baixo grau (GBG) e 592 pacientes com glioblastoma multiforme (GBM) foram adquiridos do TCGA e as alterações genéticas e os níveis de expressão dos genes selecionados foram analisados. Resultados: Identificamos seis mutações KDR distintas nos pacientes GBG e 18 mutações KDR distintas nos pacientes GBM, incluindo mutações missense e nonsense, exclusão de mudança de quadro e região de emenda alterada. Além disso, VEGFA e CXCL8 foram significativamente super-expressos nos pacientes com GBM. Conclusões: VEGFA e CXCL8 são fatores importantes para a angiogênese, os quais parecem ter um papel significativo durante a tumorigênese. Nossos resultados fornecem evidências adicionais de que o VEGFA e o CXCL8 podem induzir a angiogênese e promover o GBG a progredir no GBM. Esses achados podem ser úteis no desenvolvimento de novas abordagens terapêuticas direcionadas no futuro.
Asunto(s)
Humanos , Neoplasias Encefálicas/genética , Glioblastoma/genética , Carcinogénesis/genética , Glioma/genética , Neovascularización Patológica/genética , Valores de Referencia , Expresión Génica , Interleucina-8/análisis , Mutación Puntual/genética , Glioblastoma/patología , Receptores de Interleucina-8A/análisis , Receptores de Interleucina-8B/análisis , Receptor 2 de Factores de Crecimiento Endotelial Vascular/análisis , Factor A de Crecimiento Endotelial Vascular/análisis , Factor B de Crecimiento Endotelial Vascular/análisis , Glioma/patologíaRESUMEN
Introduction: Melanoma is the most dangerous skin cancer with high metastasis rate and mortality. Although the emergence of immunotherapy has brought hope for treatment, the mortality rate of melanoma is still increasing year by year. The underlying mechanism of melanoma tumor progression and metastasis is urgently needed to be clarified. Recently chemokines have been found to play an important role in tumor progression in addition to their immunocytochemical chemotaxis. Methods: In this study, human melanoma cell lines A375 and M14 were treated with SCH-527123, a small molecule antagonist of CXCR1 and CXCR2. The effects of treatment with SCH-527123 on melanoma cell proliferation, migration and invasion were evaluated in vitro by CCK-8, colony formation and transwell assays. Apoptosis was also detected by flow cytometry staining with annexin V and propidium iodide (PI). The molecular mechanisms of antagonist mediated were detected by western blot. Results: The results showed that SCH-527123 inhibited the proliferation, migration and invasion of melanoma cell lines and promoted apoptosis. The expression of CXCR1 and CXCR2 was downregulated after treatment with SCH-527123. PI3K/AKT pathway and downstream signaling were also inhibited at molecular level owing to treated with SCH-527123. Conclusion: In conclusion, our study demonstrated that SCH-527123, a small-molecule antagonist for CXCR1 and CXCR2 inhibited cell proliferation, migration and invasion in melanoma via PI3K/AKT pathway
Introducción: El melanoma es el cáncer de piel más peligroso, con una alta tasa de metástasis y mortalidad. Aunque la inmunoterapia ha traído esperanza para el tratamiento, la tasa de mortalidad del melanoma sigue aumentando año tras año. Es de crucial importancia aclarar el mecanismo subyacente de la evolución y la metástasis del melanoma. Recientemente se ha descubierto que las quimiocinas juegan un importante papel en la evolución tumoral. Métodos: En el presente estudio, las líneas celulares de melanoma humano A375 y M14 se trataron con SCH-527123, un inhibidor de molécula pequeña de CXCR1 y CXCR2. Los efectos del tratamiento con SCH-527123 sobre la proliferación, migración e invasión de células de melanoma se evaluaron in vitro mediante CCK-8, formación de colonias y ensayos Transwell. También se detectó apoptosis mediante citometría de flujo con tinción con anexina V y yoduro de propidio (PI). Los mecanismos moleculares del antagonista fueron detectados por Western blot. Resultados: Los resultados mostraron que SCH-527123 inhibió la proliferación, migración e invasión de líneas celulares de melanoma y promovió la apoptosis. La expresión de CXCR1 y CXCR2 disminuyó después del tratamiento con SCH-527123. La vía de señalización de la PI3K/AKT también se inhibió a nivel molecular debido a que se trataron con SCH-527123. Conclusión: Nuestro estudio demostró que SCH-527123, un inhibidor de molécula pequeña para CXCR1 y CXCR2 inhibió la proliferación celular, la migración y la invasión del melanoma a través de la vía PI3K/AKT
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Humanos , Receptores de Interleucina-8A/agonistas , Receptores de Interleucina-8B/agonistas , Invasividad Neoplásica , Melanoma/patología , Apoptosis/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/efectos de los fármacos , Antineoplásicos/farmacología , Proliferación Celular , Movimiento Celular , Células Tumorales Cultivadas , Transducción de SeñalRESUMEN
BACKGROUND@#Spinal cord injury (SCI) is a worldwide medical concern. This study aimed to elucidate the mechanism underlying the protective effect of hyperbaric oxygen (HBO) against SCI-induced neurologic defects in rats via exploring the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis and expression of brain-derived neurotrophic factor (BDNF).@*METHODS@#An acute SCI rat model was established in Sprague-Dawley rats using the Allen method. Sixty rats were divided into four groups (n = 15 in each group): sham-operated, SCI, SCI treated with HBO (SCI + HBO), and SCI treated with both HBO and AMD3100 (an antagonist of CXCR4; SCI + HBO + AMD) groups. The rats were treated with HBO twice a day for 3 days and thereafter once a day after the surgery for up to 28 days. Following the surgery, neurologic assessments were performed with the Basso-Bettie-Bresnahan (BBB) scoring system on postoperative day (POD) 7, 14, 21, and 28. Spinal cord tissues were harvested to assess the expression of SDF-1, CXCR4, and BDNF at mRNA and protein levels, using quantitative real-time polymerase chain reaction, Western blot analysis, and histopathologic analysis.@*RESULTS@#HBO treatment recovered SCI-induced descent of BBB scores on POD 14, (1.25 ± 0.75 vs. 1.03 ± 0.66, P < 0.05), 21 (5.27 ± 0.89 vs. 2.56 ± 1.24, P < 0.05), and 28 (11.35 ± 0.56 vs. 4.23 ± 1.20, P < 0.05) compared with the SCI group. Significant differences were found in the mRNA levels of SDF-1 (mRNA: day 21, SCI + HBO vs. SCI + HBO + AMD, 2.89 ± 1.60 vs. 1.56 ± 0.98, P < 0.05), CXCR4 (mRNA: day 7, SCI + HBO vs. SCI, 2.99 ± 1.60 vs.1.31 ± 0.98, P < 0.05; day 14, SCI + HBO vs. SCI + HBO + AMD, 4.18 ± 1.60 vs. 0.80 ± 0.34, P < 0.05; day 21, SCI + HBO vs. SCI, 2.10 ± 1.01 vs.1.15 ± 0.03, P < 0.05), and BDNF (mRNA: day 7, SCI + HBO vs. SCI, 3.04 ± 0.41 vs. 2.75 ± 0.31, P < 0.05; day 14, SCI + HBO vs. SCI, 3.88 ± 1.59 vs. 1.11 ± 0.40, P < 0.05), indicating the involvement of SDF-1/CXCR4 axis in the protective effect of HBO.@*CONCLUSIONS@#HBO might promote the recovery of neurologic function after SCI in rats via activating the SDF-1/CXCR4 axis and promoting BDNF expression.
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Animales , Masculino , Ratas , Western Blotting , Factor Neurotrófico Derivado del Encéfalo , Metabolismo , Modelos Animales de Enfermedad , Oxigenoterapia Hiperbárica , Métodos , Ratas Sprague-Dawley , Receptores CXCR4 , Metabolismo , Receptores de Interleucina-8A , Metabolismo , Traumatismos de la Médula Espinal , Metabolismo , TerapéuticaRESUMEN
To observe the effect of acupuncture on CXCL8 receptors (CXCR1 and CXCR2) in rat endometrium experiencing embryo implantation failure, 72 pregnant rats were randomly divided into four groups: normal group (N), embryo implantation failure group (M), acupuncture treatment group (A), and progestin treatment group (W). Then the rats in each group were equally randomized into a day-6 (D6) group, a day-8 (D8) group, and a day-10 (D10) group. The rats in group M, group A, and group W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. Meanwhile, "Housanli" and "Sanyinjiao" were selected for acupuncture. From day 1 to the time of death, the rats in group A were fastened up and then acupuncture was administered while the rats in group N and group M were only fixed, and the rats in group W were given progestin. The number of implanted embryos was calculated. The expression of CXCR1 and CXCR2 in rat endometrium was detected by immunohistochemistry, Western blotting and real-time PCR. Compared to group N, the average number of implanted embryos, the protein and mRNA expression of CXCR1 (D6, D8 and D10), and the protein and mRNA expression of CXCR2 (D8 and D10) in rat endometrium were significantly decreased in group M. Compared to group M, there was significant elevation in the average number of implanted embryos, the protein expression (D6, D8 and D10) and mRNA expression (D8) of CXCR1 in rat endometrium of group A, and the protein expression (D8 and D10) and mRNA expression (D8) of CXCR2 in rat endometrium of group W. These findings indicated that acupuncture can increase the number of implanted embryos in rats of embryo implantation failure, which may be relevant with up-regulation the expression of CXCR1 and CXCR2 at maternal-fetal interface of rats with embryo implantation failure.
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Animales , Femenino , Embarazo , Ratas , Terapia por Acupuntura , Métodos , Western Blotting , Implantación del Embrión , Genética , Endometrio , Metabolismo , Regulación del Desarrollo de la Expresión Génica , Antagonistas de Hormonas , Farmacología , Inmunohistoquímica , Mifepristona , Farmacología , Progestinas , Farmacología , Distribución Aleatoria , Ratas Wistar , Receptores de Interleucina-8A , Genética , Metabolismo , Receptores de Interleucina-8B , Genética , Metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Resultado del Tratamiento , Regulación hacia ArribaRESUMEN
CXCR1 and CXCR2 are important receptors in regulating vascular endothelial cell activities. In order to elucidate the role of CXCR1/2 in shear stress-induced endothelial cell migration, we have investigated the expression levels of CXCR1 and CXCR2 in the endothelial cells exposed to shear stress. In the experiment, anti-IL8RA and anti-IL8RB were used to antagonize CXCR1 and CXCR2. Different shear stresses were generated in a flow chamber; scratch test was carried out to compare endothelial cell migration in the control group and the receptor-antagonized groups. The results indicated that the migration of endothelial cells was restrained effectively after CXCR1 and CXCR2 were antagonized by anti-IL8RA and anti-IL8RB. And anti-IL8RA showed a stronger inhibitive effect than did anti-IL8RB (P<0.05). In the group with both receptor antagonisms, the migration was further inhibited. These results suggest that both CXCR1 and CXCR2 are important factors in mediating the migration of endothelial cells induced by shear stress, and CXCR1 fulfills a more important role.
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Humanos , Movimiento Celular , Fisiología , Células Endoteliales , Biología Celular , Metabolismo , Mecanotransducción Celular , Receptores de Interleucina-8A , Fisiología , Receptores de Interleucina-8B , Fisiología , Resistencia al Corte , Estrés Mecánico , Venas Umbilicales , Biología Celular , MetabolismoRESUMEN
Recovery from hepatitis B virus (HBV) infection depends on the cellular immune responses. Chemokines and their receptors play significant roles in immune defense. This study was undertaken to investigate the association between HBV infection and single nucleotide polymorphisms (SNPs) of genes for the chemokines and their receptors. Between March 2002 and February 2004, a total of 957 single ethnic Korean patients were enrolled into two different groups; "HBV clearance group" (n=350), who have recovered from HBV infection, and "HBV persistence group" (n=607), who were repeatedly HBsAg-positive. The HBV persistence group was subdivided into "inactive carrier" and "HBV progression group (chronic hepatitis and cirrhosis)". We assessed polymorphisms in regulated and normal T-cell expressed and secreted (RANTES) at position -403, monocyte chemoattractant protein-1 (MCP-1) at position -2518, CCR2 V64I, CCR5 -2459, CXCR1 S276T and CXCR4 I138I using single primer extension assay. Genotype distributions of the "HBV clearance versus persistence group" and "inactive carrier versus HBV progression group" were compared. On the basis of unconditional logistic regression analysis with adjustment for age and sex, no statistically significant association with susceptibility to persistent HBV infection was observed with RANTES -403, MCP-1 -2518, CCR2 V64I, CCR5 -2459, CXCR1 S276T, and CXCR4 I138I polymorphisms. In addition, no association of analyzed SNPs with HBV disease progression was found.
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Humanos , Quimiocina CCL2/genética , Quimiocina CCL5/genética , Progresión de la Enfermedad , Genotipo , Hepatitis B/etnología , Virus de la Hepatitis B/metabolismo , Corea (Geográfico) , Polimorfismo Genético , Receptores CCR2 , Receptores CCR5/genética , Receptores CXCR4/genética , Receptores de Quimiocina/genética , Receptores de Interleucina-8A/genética , Análisis de Regresión , Resultado del TratamientoRESUMEN
This study was conducted to elucidate the effects of different fluid shear stress on the expression of IL-8 receptors CXCR1 in endothelial cells. The HUVEC cell lines, EA. Hy926 cells, were cultured in vitro, exposed to 5.56, 10.02, and 15.27 dyn/cm2 laminar shear stress, respectively, and then the shear stress grous were each investigated at the time-points of 0h, 1h, 2h, 4h and 8h. Western blotting was used for detecting the expression of IL-8 receptor CXCR1 at the time-points. The results showed that, under 5.56 dyn/cm2 shear stress, the expression of CXCR1 increased with time significantly (P < 0.01). The maximum expression of CXCR1 appeared at 4 h and was 2.2 times that of the control. When exposed to 10.02 dyn/cm2, the expression of CXCR1 increased gradually with time and finally remained at a constant higher level. When exposed to 15.27 dyn/cm2, CXCR1 expression decreased significantly with time (P < 0.01). The minimum CXCR1 expression appeared at 8 h and was 62. 59 percent of that of the control. These results indicate that the expression of CXCR1 in endothelial cell is regulated by laminar shear stress.
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Humanos , Línea Celular , Células Endoteliales , Biología Celular , Metabolismo , ARN Mensajero , Genética , Receptores de Interleucina-8A , Genética , Metabolismo , Resistencia al Corte , Estrés Mecánico , Venas Umbilicales , Biología CelularRESUMEN
This study was conducted to elucidate the effects of different fluid shear stress on the expression of IL-8 receptors CXCR1 and CXCR2 mRNA in endothelial cells, EA. Hy926 cells. The HUVEC cell lines were cultured in vitro and then exposed to 5.56, 10.02, and 15.27 dyn/cm2 fluid shear stress respectively for 5 min, 10 min, 15 min, 20 min, 25 min, 30 min, 1 h, 2 h, 4 h and 8 h. Semi-quantitative reversal transcription-polymerase chain reaction (RT-PCR) was used for detecting IL-8 receptors mRNA expression at different times. The results showed that, under 5. 56 dyn/cm2 shear stress, both the expression of CXCR1 and CXCR2 mRNA increased significantly with time (P<0.05). When exposed to 10. 02 dyn/cm2, the expression of CXCR1 mRNA was down-regulated with time on every occasion. CXCR2 mRNA increased temporally at 30 min, then it decreased gradually with time and finally went on at a constant lower level. When exposed to 15.27 dyn/cm2, both CXCR1 and CXCR2 mRNA expression decreased significantly with time (P<0.01). These data indicate that the expression of CXCR1 and CXCR2 mRNA of endothelial cell is regulated by fluid shear stress.