RESUMO
Connexin26 (Cx26) plays an important role in ionizing radiation-induced damage, and CC chemokine ligand 27 (CCL27) regulates the skin immune response. However, the relationship between Cx26 and CCL27 in radiation-induced skin damage is unclear. After X-ray irradiation, clonogenic survival and micronucleus formation were assessed in immortalized human keratinocytes (HaCaT). Proteins in the mitogen activated protein kinase (MAPK) signaling pathway and CCL27-related proteins were detected by immunoblotting. HaCaTCx26-/- cells were constructed to verify the effects of Cx26 on CCL27 secretion. A mouse model was established to examine the expression of CCL27 and skin inflammation in vivo. The degree of skin injury induced by 6 MV of X rays was closely related to CCL27. The phosphorylation of ERK, p38 and NF-κB was significantly increased in irradiated cells. The secretion of CCL27 was significantly decreased in HaCaT wild-type cells relative to HaCaTCx26-/- cells. Whereas cell survival fractions decreased, and the micronuclei formation rate increased as a function of increasing X-ray dose in HaCaT cells, the opposite trend occurred in HaCaTCx26-/- cells. Our findings show that Cx26 likely plays a role in the activation of the MAPK and NF-κB/COX-2 signaling pathways and regulates the secretion of CCL27 in keratinocytes after X-ray radiation-induced skin damage.
Assuntos
Quimiocina CCL27 , Radiodermite , Animais , Humanos , Camundongos , Quimiocina CCL27/metabolismo , Quimiocina CCL27/farmacologia , Quimiocinas/metabolismo , Quimiocinas CC/metabolismo , Quimiocinas CC/farmacologia , Queratinócitos/metabolismo , Ligantes , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Proteínas Quinases Ativadas por Mitógeno/farmacologia , NF-kappa B/metabolismo , Radiodermite/etiologia , Transdução de SinaisRESUMO
Lung cancer is characterized by its high mortality and morbidity. A deep understanding of the molecular mechanisms of lung cancer tumorigenesis helps to develop novel lung cancer diagnostic and therapeutic strategies. However, the picture of the associated molecular landscape is not yet complete. As understood, chemokine-receptor interactions contribute much to lung cancer tumorigenesis, in which CCR10 also plays an important role. This study aimed to expand the knowledge of CCR10 in lung squamous cell carcinoma (LUSC) in the manner of molecular mechanism and biological functions. Using GEPIA database, the survival analysis between LUSC patients with high and low CCR10 expressions was performed, showing that CCR10 could be regarded as a risk factor for LUSC patients. Subsequently, CCR10 protein and mRNA expressions in LUSC were examined by qRT-PCR and western blot respectively. The results indicated that CCR10 was highly expressed in LUSC cells. The results of CCK-8, colony formation, and Transwell assays presented that CCL27, the ligand of CCR10, promoted proliferative, migratory, and invasive abilities of LUSC cells by activating CCR10. Also, the PI3K/AKT signaling pathway was verified as the involved pathway by western blot. Overall, it could be concluded that the CCL27-CCR10 regulatory axis can activate the PI3K/AKT pathway fostering the malignant features of LUSC cells.
Assuntos
Carcinoma Pulmonar de Células não Pequenas , Carcinoma de Células Escamosas , Neoplasias Pulmonares , Humanos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Carcinoma de Células Escamosas/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/genética , Carcinogênese/genética , Proliferação de Células , Pulmão/metabolismo , Regulação Neoplásica da Expressão Gênica , Receptores CCR10/genética , Receptores CCR10/metabolismo , Quimiocina CCL27/genética , Quimiocina CCL27/metabolismoRESUMO
BACKGROUND: In previous human skin single-cell data, inflammatory cells constituted only a small fraction of the overall cell population, such that functional subsets were difficult to ascertain. OBJECTIVE: Our aims were to overcome the aforesaid limitation by applying single-cell transcriptomics to emigrating cells from skin and elucidate ex vivo gene expression profiles of pathogenic versus regulatory immune cell subsets in the skin of individuals with psoriasis. METHODS: We harvested emigrating cells from human psoriasis skin after incubation in culture medium without enzyme digestion or cell sorting and analyzed cells with single-cell RNA sequencing and flow cytometry simultaneously. RESULTS: Unsupervised clustering of harvested cells from psoriasis skin and control skin identified natural killer cells, T-cell subsets, dendritic cell subsets, melanocytes, and keratinocytes in different layers. Comparison between psoriasis cells and control cells within each cluster revealed that (1) cutaneous type 17 T cells display highly differing transcriptome profiles depending on IL-17A versus IL-17F expression and IFN-γ versus IL-10 expression; (2) semimature dendritic cells are regulatory dendritic cells with high IL-10 expression, but a subset of semimature dendritic cells expresses IL-23A and IL-36G in psoriasis; and (3) CCL27-CCR10 interaction is potentially impaired in psoriasis because of decreased CCL27 expression in basal keratinocytes. CONCLUSION: We propose that single-cell transcriptomics applied to emigrating cells from human skin provides an innovative study platform to compare gene expression profiles of heterogenous immune cells in various inflammatory skin diseases.
Assuntos
Células Dendríticas/imunologia , Psoríase/imunologia , Pele/imunologia , Células Th17/imunologia , Movimento Celular , Separação Celular , Células Cultivadas , Quimiocina CCL27/metabolismo , Citocinas/metabolismo , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Receptores CCR10/metabolismo , Análise de Sequência de RNA , Análise de Célula Única , TranscriptomaRESUMO
OBJECTIVE: To investigate the molecular mechanism of Progranulin (PGRN) in promoting osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) in inflammatory environment. BACKGROUND: Progranulin is an antagonist of tumor necrosis factor (TNF) receptors (TNFRs) and is known to promote inflammatory periodontal bone defect regeneration. METHODS: TNFR1- and TNFR2-silenced hPDLSCs designed as hPDLSCs-sh-TNFR1 and hPDLSCs-sh-TNFR2 were cultured with osteoinductive medium containing TNF-α and (or) PGRN. Immunofluorescence, quantitative real-time PCR, and western blot were used to, respectively, detect expressions of TNFR1\TNFR2 and osteogenic differentiation markers as well as phosphorylation level in NF-κB\MAPK-related pathways. RESULTS: Immunofluorescence and real-time PCR showed that TNFR1 and TNFR2 positively expressed in hPDLSCs. TNF-α stimulation could significantly decrease the expressions of ALP and RUNX2 in hPDLSCs, whereas PGRN treatment could significantly enhance their expressions, and reverse TNF-α-mediated expression suppression of ALP and RUNX2 in hPDLSCs. In hPDLSCs-sh-TNFR1, TNF-α mediated osteogenic inhibition decreased, but both TNF-α + PGRN and alone PGRN significantly promoted expression of ALP and RUNX2. PGRN significantly enhanced expression of P-ERK1/2 and P-JNK, while corresponding inhibitors eliminated PGRN-stimulated osteogenic differentiation. In hPDLSCs-sh-TNFR2, no significant difference existed in osteogenic markers and P-JNK expression between the PGRN group and the control group. However, PGRN still activated P-ERK1/2 expression. Besides, PGRN antagonized TNF-α-enhanced NF-κB P65 expression. CONCLUSION: Progranulin promotes osteogenic differentiation of hPDLSCs via TNFR1 to inhibit TNF-α-sensitized NF-κB and via TNFR2 to activate JNK signaling. The mechanism by which PGRN activates ERK signaling remains to be explored.
Assuntos
Osteogênese , Ligamento Periodontal/citologia , Progranulinas/farmacologia , Células-Tronco/citologia , Diferenciação Celular , Células Cultivadas , Quimiocina CCL27/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases , NF-kappa B/metabolismo , Receptores do Fator de Necrose Tumoral/metabolismo , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
We compared outer and inner foreskin tissue from adolescent males undergoing medical male circumcision to better understand signals that increase HIV target cell availability in the foreskin. We measured chemokine gene expression and the impact of sexually transmitted infections (STIs) on the density and location of T and Langerhans cells. Chemokine C-C ligand 27 (CCL27) was expressed 6.94-fold higher in the inner foreskin when compared with the outer foreskin. We show that the density of CD4+CCR5+ cells/mm2 was higher in the epithelium of the inner foreskin, regardless of STI status, in parallel with higher CCL27 gene expression. In the presence of STIs, there were higher numbers of CD4+CCR5+ cells/mm2 cells in the sub-stratum of the outer and inner foreskin with concurrently higher number of CD207+ Langerhans cells (LC) in both tissues, with the latter cells being closer to the keratin surface of the outer FS in the presence of an STI. When we tested the ability of exogenous CCL27 to induce T-cell migration in foreskin tissue, CD4 + T cells were able to relocate to the inner foreskin epithelium in response. We provide novel insight into the impact CCL27 and STIs on immune and HIV-1 target cell changes in the foreskin.
Assuntos
Infecções Bacterianas/imunologia , Linfócitos T CD4-Positivos/imunologia , Quimiocina CCL27/metabolismo , Prepúcio do Pênis/metabolismo , Infecções por HIV/imunologia , HIV-1/fisiologia , Células de Langerhans/imunologia , Adolescente , Adulto , Infecções Bacterianas/terapia , Movimento Celular , Quimiocina CCL27/genética , Circuncisão Masculina , Prepúcio do Pênis/patologia , Regulação da Expressão Gênica , Infecções por HIV/terapia , Humanos , Masculino , Infecções Sexualmente Transmissíveis , África do Sul , Adulto JovemRESUMO
OBJECTIVE: To elucidate whether miRNA-27a-3p can promote osteogenic differentiation of hMSCs by targeting ATF3, thus alleviating osteoporosis symptoms. PATIENTS AND METHODS: The serum levels of miRNA-27a-3p in osteoporosis patients (n=20) and normal controls (n=20) were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Human bone marrow mesenchymal stem cells (hMSCs) were subjected to osteogenic differentiation for 1, 3 and 7 days. Subsequently, mRNA levels of miRNA-27a-3p, ALP, and Bglap in hMSCs were determined by qRT-PCR. The regulatory effects of miRNA-27a-3p levels and the mRNA levels of ALP, Bglap, and Runx2 were detected. After the overexpression or knockdown of miRNA-27a-3p, we evaluated the changes in the osteogenic differentiation by alizarin red staining and ALP staining. Through Dual-Luciferase Reporter Gene Assay, we verified the binding relationship between miRNA-27a-3p and ATF3. Rescue experiments were finally conducted to prove whether miRNA-27a-3p regulated the osteogenic differentiation by targeting ATF3. RESULTS: The serum level of miRNA-27a-3p remained lower in osteoporosis patients relative to controls. With the prolongation of osteogenic differentiation, the mRNA levels of miRNA-27a-3p, ALP, and Bglap gradually increased. The overexpression of miRNA-27a-3p upregulated mRNA and the protein levels of osteogenesis-related genes, increased ALP activity, and enhanced mineralization capacity. The knockdown of miRNA-27a-3p obtained the opposite trends. MiRNA-27a-3p could target ATF3, and the overexpression of ATF3 reversed the promotive effects of miRNA-27a-3p on osteogenic differentiation. CONCLUSIONS: MiRNA-27a-3p promotes the differentiation of hMSCs into osteoblasts by targeting ATF3, thus alleviating osteoporosis symptoms.
Assuntos
Fator 3 Ativador da Transcrição/genética , Células-Tronco Mesenquimais/citologia , MicroRNAs/genética , Osteoporose/genética , Regiões 3' não Traduzidas , Fator 3 Ativador da Transcrição/metabolismo , Estudos de Casos e Controles , Diferenciação Celular , Células Cultivadas , Quimiocina CCL27/metabolismo , Regulação para Baixo , Regulação da Expressão Gênica , Humanos , Células-Tronco Mesenquimais/metabolismo , Osteogênese , Osteoporose/metabolismoRESUMO
The appearance of mammalian-specific skin features was a key evolutionary event contributing for the elaboration of physiological processes such as thermoregulation, adequate hydration, locomotion, and inflammation. Skin inflammatory and autoimmune processes engage a population of skin-infiltrating T cells expressing a specific C-C chemokine receptor (CCR10) which interacts with an epidermal CC chemokine, the skin-specific C-C motif chemokine ligand 27 (CCL27). CCL27 is selectively produced in the skin by keratinocytes, particularly upon inflammation, mediating the adhesion and homing of skin-infiltrating T cells. Here, we examined the evolution and coding condition of Ccl27 in 112 placental mammalian species. Our findings reveal that a number of open reading frame inactivation events such as insertions, deletions, and start and stop codon mutations independently occurred in Cetacea, Pholidota, Sirenia, Chiroptera, and Rodentia, totalizing 18 species. The diverse habitat settings and lifestyles of Ccl27-eroded lineages probably implied distinct evolutionary triggers rendering this gene unessential. For example, in Cetacea, the rapid renewal of skin layers minimizes the need for an elaborate inflammatory mechanism, mirrored by the absence of epidermal scabs. Our findings suggest that the convergent and independent loss of Ccl27 in mammalian evolution concurred with unique adaptive roads for skin physiology.
Assuntos
Quimiocina CCL27/genética , Evolução Molecular , Inativação Gênica , Pele/metabolismo , Sequência de Aminoácidos , Animais , Cetáceos/genética , Quimiocina CCL27/química , Quimiocina CCL27/metabolismo , Quimiocinas CC/química , Quimiocinas CC/genética , Quimiocinas CC/metabolismo , Éxons , Feminino , Loci Gênicos , Predisposição Genética para Doença , Humanos , Mamíferos , Modelos Moleculares , Mutação , Splicing de RNA , Análise de Sequência de DNA , Relação Estrutura-Atividade , Transcrição Gênica , TranscriptomaRESUMO
INTRODUCTION: Multiple sclerosis (MS) is a neuroinflammatory autoimmune disease which involves the central nervous -system. Although the primary cause of MS is obscure, effects of some cytokine and chemokine patterns in both innate and adaptive immune systems have been described. -Objectives: Since limited studies have examined the role of interleukin (IL)-11 and chemokine CCL27 in MS, we aimed to identify changes in IL-11 and CCL27 gene expression and serum levels in relapsing-remitting MS (RRMS) patients, treated with interferon (IFN)-ß and glatiramer acetate (GA). METHODS: The serum level and gene expression of IL-11 and CCL27 were measured and compared between treatment-naïve MS patients and RRMS patients who were treated with high-dose IFN-ß1a, low-dose IFN-ß1a, IFN-ß1b, and GA via enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction. RESULTS: A significant decrease was observed in the serum level of CCL27 in treatment-naïve patients and IFN-ß1b-treated patients compared to the healthy controls. On the other hand, a significant increase was found in the protein level of CCL27 in low-dose and high-dose IFN-ß1a groups compared to the treatment-naïve group. In addition, CCL27 gene expression was higher in patients treated with GA than in the treatment-naïve group. There were no significant changes in the gene expression or protein level of IL-11 in all experimental groups. Additionally, a positive correlation was found between IL-11 and CCL-27. CONCLUSION: Our results suggest the inflammatory role of CCL27 in MS patients, while IFN-ß1a seems to play a compensatory role for this chemokine.
Assuntos
Quimiocina CCL27/metabolismo , Acetato de Glatiramer/uso terapêutico , Imunossupressores/uso terapêutico , Interferon beta-1b/uso terapêutico , Interleucina-11/metabolismo , Esclerose Múltipla Recidivante-Remitente/imunologia , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológicoRESUMO
Keloid scars are often described as having an actively growing peripheral margin with a regressing centre. The aim of this study was to examine the possible heterogeneity within keloids and the involvement of different regions within and around keloid scars in the pathogenesis, using an in vitro keloid scar model. In vitro skin models were constructed from keratinocytes and fibroblasts from normal skin and different regions within and around keloid scars: periphery, centre, and (adjacent) surrounding-normal-skin regions. Additionally, fibroblasts were isolated from the superficial-central and deep-central regions of the keloid and combined with central keratinocytes. All keloid regions showed increased contraction compared to normal skin models, particularly in central regions. Myofibroblasts were present in all keloid regions but were more abundant in models containing central-deep keloid fibroblasts. Secretion of anti-fibrotic HGF and extracellular matrix collagen IV gene expression was reduced in the central deep keloid compared to normal skin. No significant differences between peripheral and central regions within keloids were observed for inflammatory cytokine CCL20, CCL27, CXCL8, IL-6 and IL-18 secretion. Parameters for surrounding-normal-skin showed similarities to both non-lesional normal skin and keloids. In conclusion, a simple but elegant method of culturing keloid-derived keratinocytes and fibroblasts in an organotypic 3D scar model was developed, for the dual purpose of studying the underlying pathology and ultimately testing new therapeutics. In this study, these tissue engineered scar models show that the central keloid region shows a more aggressive keloid scar phenotype than the periphery and that the surrounding-normal-skin also shares certain abnormalities characteristic for keloids.
Assuntos
Proliferação de Células/fisiologia , Cicatriz Hipertrófica/patologia , Fibroblastos/metabolismo , Queloide/patologia , Queratinócitos/metabolismo , Pele/patologia , Quimiocina CCL20/metabolismo , Quimiocina CCL27/metabolismo , Criança , Pré-Escolar , Colágeno/metabolismo , Feminino , Fator de Crescimento de Hepatócito/metabolismo , Humanos , Lactente , Interleucina-18/metabolismo , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Masculino , Miofibroblastos/metabolismoRESUMO
Diabetic wound care is a major health care concern. The major cause of non-healing of wounds in patients with diabetes mellitus (DM) patients mainly involves poor glycemic control, which hinders the migration of progenitor cells including mesenchymal stem cells to the wound site. In this study, we introduced adipose-derived stromal cells (ADSCs) into wound sites and demonstrated that the local transplantation of ADSCs accelerated DM-related wound healing. Furthermore, the migration ability of ADSCs, which diminishes in a high-glucose environment, was partially restored by the exogenous replenishment of the cutaneous T-cell attracting chemokine (CTACK/CCL27). Our findings suggest that CTACK is a potential novel therapeutic target in DM-related wound healing.
Assuntos
Tecido Adiposo/citologia , Glicemia/metabolismo , Movimento Celular/fisiologia , Quimiocina CCL27/metabolismo , Diabetes Mellitus/sangue , Diabetes Mellitus/metabolismo , Células Estromais/citologia , Cicatrização/fisiologia , Animais , Células Cultivadas , Humanos , Masculino , Ratos , Ratos WistarRESUMO
The hybrid training system (HTS) is a special and compact system for effective skeletal muscle training by a combined application of volitional and electrical muscle contraction. Lower limbs' muscle training using HTS has been reported to increase not only muscle strength but also plasma interleukin-6 levels; however, little is known in other cytokines. In this study, we measured 52 cytokines and creatine phosphokinase-MM in the serum of 16 healthy men before and after lower limbs' muscle training by the knee flexion and extension using HTS. Skeletal muscle volume-corrected serum concentrations of cutaneous T-cell-attracting chemokine, erythropoietin, and tumor necrosis factor-related apoptosis-inducing ligand increased immediately after the training. These increased cytokines have been reported to play important roles in wound healing, neuroprotection, and cardiovascular protection.
Assuntos
Citocinas/metabolismo , Músculo Esquelético/metabolismo , Adulto , Apoptose/fisiologia , Quimiocina CCL27/metabolismo , Eritropoetina/metabolismo , Humanos , Joelho/fisiologia , Extremidade Inferior/fisiologia , Masculino , Contração Muscular/fisiologia , Força Muscular/fisiologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The existing therapies of IgA nephropathy are unsatisfying. Acteoside, the main component of Rehmannia glutinosa with anti-inflammatory and anti-immune effects, can improve urinary protein excretion and immune disorder. Th22 cell is involved in IgA nephropathy progression. This study was determined to explore the effect of acteoside on mesangial injury underlying Th22 cell disorder in IgA nephropathy. Serum Th22 cells and urine total protein of patients with IgA nephropathy were measured before and after six months treatment of Rehmannia glutinosa acteoside or valsartan. Chemotactic assay and co-culture assay were performed to investigate the effect of acteoside on Th22 cell chemotaxis and differentiation. The expression of CCL20, CCL22 and CCL27 were analyzed. To explore the effect of acteoside on mesangial cell injury induced by inflammation, IL-1, IL-6, TNF-α and TGF-ß1 were tested. Results showed that the proteinuria and Th22 lymphocytosis of patients with IgA nephropathy significantly improved after combination treatment of Rehmannia glutinosa acteoside and valsartan, compared with valsartan monotherapy. In vitro study further demonstrated that acteoside inhibit Th22 cell chemotaxis by suppressing the production of Th22 cell attractive chemokines, i.e., CCL20, CCL22 and CCL27. In addition, acteoside inhibited the Th22 cell proliferation. Co-culture assay proved that acteoside could relieve the overexpression of pro-inflammatory cytokines, and prevent the synthesis of TGF-ß1. TGF-ß1 level in mesangial cells was positively correlated with the Th22 cell. This research demonstrated that acteoside can alleviate mesangial cell inflammatory injury by modulating Th22 lymphocytes chemotaxis and proliferation.
Assuntos
Glomerulonefrite por IGA/tratamento farmacológico , Glucosídeos/farmacologia , Imunossupressores/farmacologia , Células Mesangiais/efeitos dos fármacos , Fenóis/farmacologia , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos , Adulto , Biópsia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Quimiocina CCL20/imunologia , Quimiocina CCL20/metabolismo , Quimiocina CCL22/imunologia , Quimiocina CCL22/metabolismo , Quimiocina CCL27/imunologia , Quimiocina CCL27/metabolismo , Quimiotaxia/efeitos dos fármacos , Quimiotaxia/imunologia , Técnicas de Cocultura , Progressão da Doença , Quimioterapia Combinada/métodos , Feminino , Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Glucosídeos/uso terapêutico , Humanos , Imunossupressores/uso terapêutico , Masculino , Células Mesangiais/imunologia , Células Mesangiais/patologia , Pessoa de Meia-Idade , Fenóis/uso terapêutico , Proteinúria/tratamento farmacológico , Rehmannia/química , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Fator de Crescimento Transformador beta1/imunologia , Fator de Crescimento Transformador beta1/metabolismo , Resultado do Tratamento , Valsartana/uso terapêutico , Adulto JovemRESUMO
The skin-associated chemokine CTACK plays a key role in many inflammatory conditions and could be instrumental in the pathophysiology of tissue-specific immunological diseases such as oral lichen planus (OLP). In this study, we investigated, by RT-PCR, ELISA, chemotaxis assays, and fluorescence-activated cell sorting (FACS), the production of CTACK in oral keratinocytes, its expression in tissues from normal and OLP patients, and its role in T-cell recruitment.CTACK was produced by the oral epithelium, and it affects chemotaxis of memory CLA+ cells to the oral epithelium. CTACK mRNA was expressed constitutively in primary oral epithelium and was increased during pro-inflammatory IFN-γ treatment. We found a constitutive production of CTACK at a protein level in oral primary cells that increased after IFN-γ treatment. Moreover, we confirmed that CTACK attracts memory T cells and those T cells that express CLA above the level of basal migration.
Assuntos
Quimiocina CCL27/metabolismo , Queratinócitos/citologia , Mucosa Bucal/metabolismo , Linhagem Celular , Movimento Celular , Separação Celular , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Inflamação , Interferon gama/metabolismo , Líquen Plano Bucal/metabolismo , RNA Mensageiro/metabolismo , Proteínas Recombinantes/metabolismoRESUMO
CCR10, a member of the chemokine receptor subfamily, is overexpressed in several tumors and play a crucial role in cancer development and progression. However, the functions of CCR10 in breast cancer are unknown. Here, we detected the protein levels of CCR10 in breast cancer cells by western blotting, and examined CCR10 expression in breast cancer tissues via immunohistochemical assay. The results showed that CCR10 expression was elevated in breast cancer MCF7, BT-474 and MDA-MB-231 cells. Further, 63 of 89 cases (70.8%) had positive CCR10 staining, and the CCR10 level was closely related to capsular invasion, lymph node metastasis and tumor stage. Moreover, CCL27, the ligand of CCR10, dose-dependently stimulated the invasion and migration of breast cancer cells, and promoted MMP-7 expression and ERK1/2 activation. CCR10 knockdown in breast cancer cells through siRNA transfection attenuated CCL27-induced cell invasiveness, and suppressed MMP-7 expression and ERK1/2 activation. Additionally, blocking the ERK1/2 pathway inhibited the CCL27/CCR10-promoted cell invasion of breast cancer cells. Together, these data suggest that CCL27/CCR10 interaction induces the ERK1/2 pathway, which then increases MMP-7 expresion and subsequently promotes breast cancer cell invasion and migration. Thus, CCR10 may be a key regulator in breast cancer cell invasion and migration.
Assuntos
Neoplasias da Mama/imunologia , Metaloproteinase 7 da Matriz/metabolismo , Receptores CCR10/metabolismo , Neoplasias da Mama/patologia , Carcinogênese , Movimento Celular , Quimiocina CCL27/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Sistema de Sinalização das MAP Quinases , Células MCF-7 , Pessoa de Meia-Idade , Estadiamento de Neoplasias , RNA Interferente Pequeno/genética , Receptores CCR10/genéticaRESUMO
Angiotensin II is considered a major profibrotic factor that is involved in tissue remodeling processes, as the inhibition of Angiotensin II can halt renal inflammatory processes. Dexamethasone, an important anti-inflammatory and immunosuppressive agent, has been widely used to treat renal disease for decades. In this study, we explored the frequency of Th22 cells in a mouse model of IgA nephropathy and compared the possible effects of Losartan and Dexamethasone on Th22 cells. The experiments were performed using 6-week-old BALB/c female mice in an established IgA nephropathy model. The mice were randomly separated into 4 groups, which were administered Losartan (30mg/kg/d) or Dexamethasone (10mg/kg/d) and subjected to IgA nephropathy or the normal control treatment for 1month. The frequency of Th22 cells was measured via flow cytometry, and the relative pathological changes in renal morphology were measured with different pathological staining methods. Immunohistochemistry was performed to verify the expression of CCR10 and CCL27, which is specialized receptor on Th22 cells and its corresponding chemokine, respectively. The concentrations of CCL27 and IL-22 in renal tissue homogenates and sera were detected using ELISAs. Losartan and Dexamethasone differentially decreased the frequency of Th22 cells after 1month, and mesangial cell proliferation was also improved. Moreover, the expression of CCR10, CCL27 and IL-22 was reduced by treatment with either drug. However, significant differences between Losartan and Dexamethasone were not observed. Based on these findings, Losartan and Dexamethasone may suppress inflammatory responses by inhibiting the chemotaxis of Th22 cells in IgA nephropathy.
Assuntos
Quimiocina CCL27/metabolismo , Dexametasona/uso terapêutico , Glomerulonefrite por IGA/tratamento farmacológico , Imunossupressores/uso terapêutico , Interleucinas/metabolismo , Losartan/uso terapêutico , Receptores CCR10/metabolismo , Linfócitos T Auxiliares-Indutores/efeitos dos fármacos , Animais , Células Cultivadas , Quimiotaxia/efeitos dos fármacos , Feminino , Glomerulonefrite por IGA/imunologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Linfócitos T Auxiliares-Indutores/imunologia , Interleucina 22RESUMO
The skin-associated chemokine CCL27 and its receptor CCR10 mediate the immune response of skin-homing T cells. The CCL27 secreted from keratinocytes was reportedly involved in inflammatory skin diseases such as atopic dermatitis, contact dermatitis, and psoriasis. However, whether ionizing radiation increases the levels of CCL27 secretion still remains unclear. In HaCaT cells, a human keratinocyte cell line, CCL27 secretion was markedly increased after X-ray irradiation. We further found that irradiation boosted the generation of reactive oxygen species (ROS), which was concomitant with the release of tumor necrosis factor-alpha (TNF-α). Moreover, alteration of ROS in irradiated HaCaT cells correlated with TNF-α secretion, indicating a positive loop of TNF-α secretion and ROS generation. This positive loop regulated the secretion of CCL27 from irradiated cells. We therefore concluded that the cross talk between TNF-α and ROS after keratinocytes was exposed to radiation, triggered CCL27 secretion for subsequent inflammation response.
Assuntos
Quimiocina CCL27/metabolismo , Imunidade Celular/efeitos dos fármacos , Inflamação/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Dermatite de Contato/imunologia , Dermatite de Contato/metabolismo , Dermatite de Contato/patologia , Humanos , Inflamação/imunologia , Inflamação/patologia , Queratinócitos/metabolismo , Queratinócitos/efeitos da radiação , NF-kappa B/metabolismo , Psoríase/imunologia , Psoríase/metabolismo , Psoríase/patologia , Radiação Ionizante , Receptores CCR10/metabolismo , Linfócitos T/imunologia , Linfócitos T/efeitos da radiaçãoRESUMO
AIMS: It has been proposed that the expression of chemokines and chemokine receptors by melanoma cells may have a role in tumour immune escape. Chemokine CCL27 is reported to be expressed specifically on the epidermal keratinocytes. The implication of CCL27 in cutaneous melanomas is currently unresolved. It has been suggested that CCL27 expression in melanomas can induce antitumoral immunity, and that CCL27 may suppress tumour growth probably due to the local lymphocyte recruitment. METHODS: We studied CCL27 chemokine expression in three different concentric epidermal areas covering the primary cutaneous melanoma in patients with a long clinical follow-up. Our study included 91 cases of primary melanomas of the skin diagnosed during the 10-year period 1992-2002, and a minimum clinical follow-up of 10â years. RESULTS: We evaluated three different concentric and easily reproducible areas in the epidermis: the area covering melanoma (which we called 'supratumoral'), the area adjacent to the tumour ('peritumoral') and the most peripheral epidermal area ('peripheral'). Only CCL27 expression in supratumoral epidermis correlated with clinical outcome. CONCLUSIONS: Our study showed that a higher immunostaining of CCL27 in supratumoral epidermis is associated with longer progression-free interval and melanoma-specific survival.
Assuntos
Quimiocina CCL27/metabolismo , Epiderme/metabolismo , Melanoma/metabolismo , Neoplasias Cutâneas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Intervalo Livre de Doença , Epiderme/patologia , Feminino , Humanos , Imuno-Histoquímica , Queratinócitos/metabolismo , Queratinócitos/patologia , Masculino , Melanoma/mortalidade , Melanoma/patologia , Pessoa de Meia-Idade , Prognóstico , Neoplasias Cutâneas/mortalidade , Neoplasias Cutâneas/patologia , Taxa de Sobrevida , Adulto Jovem , Melanoma Maligno CutâneoRESUMO
The bone marrow microenvironment plays a decisive role in multiple myeloma progression and drug resistance. Chemokines are soluble mediators of cell migration, proliferation and survival and essentially modulate tumor progression and drug resistance. Here we investigated bone marrow-derived chemokines of naive and therapy-refractory myeloma patients and discovered that high levels of the chemokine CCL27, known so far for its role in skin inflammatory processes, correlated with worse overall survival of the patients. In addition, chemokine levels were significantly higher in samples from patients who became refractory to bortezomib at first line treatment compared to resistance at later treatment lines.In vitro as well as in an in vivo model we could show that CCL27 triggers bortezomib-resistance of myeloma cells. This effect was strictly dependent on the expression of the respective receptor, CCR10, on stroma cells and involved the modulation of IL-10 expression, activation of myeloma survival pathways, and modulation of proteasomal activity. Drug resistance could be totally reversed by blocking CCR10 by siRNA as well as blocking IL-10 and its receptor.From our data we suggest that blocking the CCR10/CCL27/IL-10 myeloma-stroma crosstalk is a novel therapeutic target that could be especially relevant in early refractory myeloma patients.
Assuntos
Bortezomib/farmacologia , Quimiocina CCL27/metabolismo , Resistencia a Medicamentos Antineoplásicos , Mieloma Múltiplo/tratamento farmacológico , Complexo de Endopeptidases do Proteassoma/metabolismo , Inibidores de Proteassoma/farmacologia , Receptor Cross-Talk/efeitos dos fármacos , Receptores CCR10/metabolismo , Transdução de Sinais/efeitos dos fármacos , Idoso , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Feminino , Humanos , Interleucina-10/genética , Interleucina-10/metabolismo , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/enzimologia , Mieloma Múltiplo/genética , Mieloma Múltiplo/patologia , Interferência de RNA , Receptores CCR10/genética , Receptores de Interleucina-10/genética , Receptores de Interleucina-10/metabolismo , Transfecção , Microambiente Tumoral , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Novel specific therapies for psoriasis and eczema have been developed, and they mark a new era in the treatment of these complex inflammatory skin diseases. However, within their broad clinical spectrum, psoriasis and eczema phenotypes overlap making an accurate diagnosis impossible in special cases, not to speak about predicting the clinical outcome of an individual patient. Here, we present a novel robust molecular classifier (MC) consisting of NOS2 and CCL27 gene that diagnosed psoriasis and eczema with a sensitivity and specificity of >95% in a cohort of 129 patients suffering from (i) classical forms; (ii) subtypes; and (iii) clinically and histologically indistinct variants of psoriasis and eczema. NOS2 and CCL27 correlated with clinical and histological hallmarks of psoriasis and eczema in a mutually antagonistic way, thus highlighting their biological relevance. In line with this, the MC could be transferred to the level of immunofluorescence stainings for iNOS and CCL27 protein on paraffin-embedded sections, where patients were diagnosed with sensitivity and specificity >88%. Our MC proved superiority over current gold standard methods to distinguish psoriasis and eczema and may therefore build the basis for molecular diagnosis of chronic inflammatory skin diseases required to establish personalized medicine in the field.
Assuntos
Quimiocina CCL27/metabolismo , Eczema/diagnóstico , Óxido Nítrico Sintase Tipo II/metabolismo , Psoríase/diagnóstico , Adulto , Idoso , Estudos de Coortes , Eczema/classificação , Eczema/metabolismo , Feminino , Imunofluorescência , Humanos , Masculino , Pessoa de Meia-Idade , Psoríase/classificação , Psoríase/metabolismoRESUMO
Atopic dermatitis (AD) is a chronic relapsing eczematous skin disease. Certain populations of patients are resistant to standard therapies with topical steroids and/or calcineurin inhibitors, and require systemic medication, such as immunosuppressants. Recently, several reports have shed light on the anti-allergic effects of carotenoids. Therefore, we investigated the effect of p.o. administration of ß-carotene or lycopene on AD-like symptoms of HR-1 hairless mice fed with a low zinc/magnesium diet. Mice were divided into four groups: (i) fed with a standard diet (Co group); (ii) low zinc/magnesium diet (HR group); (iii) low zinc/magnesium and ß-carotene diet (HR-C group); and (iv) low zinc/magnesium and lycopene diet (HR-L group). They were then fed these diets for 8 weeks. Severities of dermatitis were assessed by their appearance, and histopathological and hematological observations. Mice in the HR group developed AD-like dermatitis both clinically and histologically. HR-C and HR-L group mice also developed xerosis and wrinkle-like skin changes, but they were milder than those of HR group mice. Histological analysis revealed that epidermis thickening and inflammatory cell infiltration in the skin of the HR-C and HR-L groups were both statistically less than those of the HR group. The concentration of thymus and activation regulated chemokine in the skin of the HR-L group and the concentration of CCL27 in the skin of the HR-C group were significantly lower than those of the HR group, respectively. In conclusion, p.o. administration of ß-carotene or lycopene prevents AD-like symptoms in association with a suppression of T-helper 2 chemokines in a murine model. Ingestion of carotenoids may be beneficial for patients with AD.