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Tumor-infiltrating immune cells engage in an extensive crosstalk with tumors and act as two-edged swords by inhibiting or promoting cancer growth. Therefore, identifying the density and prognostic values of tumor-infiltrating immune cells will provide valuable tips for cancer treatments. In this study, we identified the density of tumor inflammatory infiltrates and the number of tumor-infiltrating immune cells, including CD3+, CD4+, CD8+, FoxP3+ T cells and CD1a+ dendritic cells (DCs) in 153 tongue squamous cell carcinomas (TSCC). High inflammatory cell infiltration was associated with better overall survival (OS) and disease free survival (DFS). Moreover, the number of CD3+, CD4+, FoxP3+ and CD1a+ cells were associated with tumor differentiation (P<0.001) and the number of FoxP3+, CD1a+ cells and CD8+/FoxP3+ ratios were also associated with tumor stage (P<0.01, P<0.01, P<0.05). In addition, patients with higher CD1a+ DCs had better OS and increased CD8+/FoxP3+ ratios were associated with improved OS and DFS (P = 0.037; P = 0.047; P = 0.033). In conclusion, our results indicated that tumor-infiltrating CD1a+ DCs and CD8+/FoxP3+ ratios were associated with favorable clinical outcomes but not independent prognostic factors for TSCC patients.
Assuntos
Células Dendríticas/imunologia , Linfócitos do Interstício Tumoral/imunologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/imunologia , Subpopulações de Linfócitos T/imunologia , Neoplasias da Língua/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Neoplasias da Língua/patologia , Adulto JovemRESUMO
C-type lectin receptors (CLRs) are powerful pattern-recognition receptors that discern 'self' and 'non-self' in our body and protect us from invasive pathogens by mediating immune recognition and response. The gastrointestinal tract is very important for the maintenance of homeostasis; it is the largest shelter for the billions of microorganisms in the body and CLRs play a crucial regulatory role in this system. This study focuses on several CLRs, including Dectin-1, Dectin-2, Dectin-3 and Mincle. We summarize the roles of CLRs in maintaining gastrointestinal immune-system homeostasis, especially their functions in mediating immune recognition and responses in the gut, discuss their relationships to some diseases, highlight the significance of CLR-mediated sensing of microbial and non-microbial compounds in the gut immune system and identify new therapeutic targets.
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Sepsis is a life-threatening health condition that is initially characterized by uncontrolled inflammation, followed by the development of persistent immunosuppression. YCP is a novel α-glucan purified from the mycelium of the marine fungus Phoma herbarum YS4108, which has displayed strong antitumor activity via enhancing host immune responses. In this study, we investigated whether YCP could influence the development of sepsis in a mouse model. Caecal ligation and puncture (CLP)-induced sepsis was established in mice that were treated with YCP (20 mg/kg, ip or iv) 2 h before, 4 and 24 h after the CLP procedure, and then every other day. YCP administration greatly improved the survival rate (from 39% to 72% on d 10 post-CLP) and ameliorated disease symptoms in the septic mice. Furthermore, YCP administration significantly decreased the percentage of myeloid-derived suppressor cells (MDSCs) in the lungs and livers, which were dramatically elevated during sepsis. In cultured BM-derived cells, addition of YCP (30, 100 µg/mL) significantly decreased the expansion of MDSCs; YCP dose-dependently decreased the phosphorylation of STAT3 and increased the expression of interferon regulatory factor-8 (IRF-8). When BM-derived MDSCs were co-cultured with T cells, YCP dose-dependently increased the production of arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS), and activated the NF-κB pathway. In addition, the effects of YCP on MDSCs appeared to be dependent on toll-like receptor (TLR) 4. These results reveal that YCP inhibits the expansion of MDSCs via STAT3 while enhancing their immunosuppressive function, partially through NF-κB. Our findings suggest that YCP protects mice against sepsis by regulating MDSCs. Thus, YCP may be a potential therapeutic agent for sepsis.
Assuntos
Células Supressoras Mieloides/efeitos dos fármacos , Polissacarídeos/farmacologia , Choque Séptico/tratamento farmacológico , Animais , Ascomicetos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Células Supressoras Mieloides/metabolismo , Células Supressoras Mieloides/patologia , Polissacarídeos/química , Polissacarídeos/isolamento & purificação , Choque Séptico/metabolismo , Choque Séptico/patologia , Relação Estrutura-Atividade , Taxa de Sobrevida , SíndromeRESUMO
AIMS: Toll-like receptor (TLR)-7 agonists have been used in cancer immunotherapy, but tumour heterogeneity means that TLR-7 activity is variable in different components of the tumour microenvironment and the characteristics of TLR-7 in oral squamous cell carcinoma (OSCC) are unclear. METHODS AND RESULTS: Twenty healthy oral tissues, 50 oral leukoplakia tissues and 166 retrospective primary OSCC samples were collected for immunohistochemical staining of TLR-7 and showed up-regulated expression during carcinogenesis. Moreover, patients with high expression of TLR-7 in tumour cells (TCs) had poor differentiation and prognosis. Interestingly, patients with high expression of TLR-7 in stroma fibroblast-like cells (FLCs) had low tumour stage, no lymph node metastasis (LNM) and better prognosis. Furthermore, Ki-67, CD3, CD4, CD8 and forkhead box protein 3 (FoxP3)(+) tumour-infiltrated lymphocytes were assessed and we found that TLR-7(high) TCs were infiltrated by fewer CD3(+) CD4(+) but more FoxP3(+) lymphocytes. Importantly, patients with TLR-7(low) TCs and TLR-7(high) FLCs had less FoxP3(+) lymphocyte infiltration and longer survival time than those with TLR-7(high) TCs/TLR-7(low) FLCs, although TLR-7 was not an independent prognostic factor for OSCC. CONCLUSIONS: The low expression of TLR-7 in tumour and high expression of TLR-7 in stroma predict a good clinical outcome for OSCC patients, and stroma FLCs might be amenable to immunotherapy by a TLR-7 agonist.
Assuntos
Biomarcadores Tumorais/análise , Carcinoma de Células Escamosas/patologia , Neoplasias de Cabeça e Pescoço/patologia , Neoplasias Bucais/patologia , Receptor 7 Toll-Like/biossíntese , Idoso , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/mortalidade , Intervalo Livre de Doença , Feminino , Fibroblastos/metabolismo , Fibroblastos/patologia , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/mortalidade , Humanos , Imuno-Histoquímica , Estimativa de Kaplan-Meier , Linfócitos do Interstício Tumoral/patologia , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/metabolismo , Neoplasias Bucais/mortalidade , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Carcinoma de Células Escamosas de Cabeça e Pescoço , Células Estromais/metabolismo , Células Estromais/patologia , Receptor 7 Toll-Like/análise , Microambiente Tumoral/fisiologiaRESUMO
INTRODUCTION: Previous studies have produced controversial results regarding whether mesenchymal stem cells (MSCs) promote or inhibit tumor development. Given the dual role of MSCs in inflammation and cancer, in this study the colitis-associated colorectal cancer (CAC) model was used to examine whether umbilical cord tissue-derived MSCs could prevent neoplasm by inhibiting chronic inflammation. METHODS: MSCs were obtained and identified using flow cytometry. Colitis-associated colorectal cancer model was induced using azoxymethane (AOM) and dextran sulfate sodium (DSS) and MSCs were injected intravenously twice. Levels of immune cells in mesenteric lymph node including regulatory T (Treg) cells were detected using flow cytometry. Naïve T cells and Jurkat cells were co-cultured with MSCs and the effect of MSCs on Treg cells differentiation was evaluated. RESULTS: After injection through tail vein, MSCs could migrate to colon and suppress colitis-related neoplasm. This tumor suppressive effect was characterized by longer colon length, decreased tumor numbers and decreased expression of Ki-67. Moreover, MSCs alleviated the pathology of inflammation in the colitis stage of CAC model and inhibited inflammation cytokines both in colon and serum. Furthermore, Treg cells were accumulated in mesenteric lymph node of MSCs-treated mice while the percentage of T helper cells 2 (Th2) and Th17 were not changed. Of note, MSCs secreted transforming growth factor-ß (TGF-ß) enhanced the induction of Treg cells from naïve T cells. The conditioned medium of MSCs also activated Smad2 signaling, which has been reported to regulate Treg cells. CONCLUSIONS: These results proved that MSCs could migrate to colon tissues and induce the differentiation of Treg cells via Smad2 as so to inhibit the colitis and suppress the development of CAC.
Assuntos
Colite/patologia , Neoplasias Colorretais/prevenção & controle , Células-Tronco Mesenquimais/metabolismo , Proteína Smad2/metabolismo , Linfócitos T Reguladores/imunologia , Animais , Azoximetano , Contagem de Linfócito CD4 , Diferenciação Celular , Linhagem Celular , Movimento Celular , Terapia Baseada em Transplante de Células e Tecidos/métodos , Colite/induzido quimicamente , Colite/imunologia , Colo/citologia , Colo/imunologia , Colo/patologia , Neoplasias Colorretais/patologia , Citocinas/sangue , Sulfato de Dextrana , Modelos Animais de Doenças , Humanos , Inflamação/imunologia , Inflamação/patologia , Inflamação/terapia , Células Jurkat , Ativação Linfocitária/imunologia , Contagem de Linfócitos , Masculino , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos C57BL , Transdução de Sinais , Linfócitos T Reguladores/citologia , Células Th17/imunologia , Células Th2/imunologia , Cordão Umbilical/citologiaRESUMO
CD68 has been widely used as a pan-macrophage marker for tumor-associated macrophages (TAM) which always involve in carcinogenesis. But the correlations between CD68(+) TAMs and prognosis of patients show to be inconsistent, which might due to lack of specific markers of TAMs. We here found that the microlocalization of CD68(+) TAMs also played a unique role in prognosis of patients with oral squamous cell carcinoma (OSCC). CD68(+) TAMs were identified in paraffin-embedded OSCC specimens (n = 91) by using immunohistochemistry. The number of CD68(+) TAMs was remarkably increased from adjacent none-neoplasia tissues (NT) to tumor nest (TN), but tumor stroma (TS) was infiltrated with highest frequency of CD68(+) TAMs (P < 0.0001). Unexpectedly, more CD68(+) TAMs in TS, but not NT or TN, were associated with high tumor grade (P = 0.033), lymph node metastasis (P = 0.034), and shorter 10-year overall survival time, disease free survival. Considering TAMs was derived from monocytes in peripheral blood, we assessed the relationship between leukocytes in peripheral blood and CD68(+) TAMs in OSCC and found that more CD68(+) TAMs in TS were accompanied with decreased monocytes and lymphocytes in peripheral blood (P < 0.05). Although Cox regression analysis revealed that CD68(+) TAMs in TS were not an independent prognostic factor for OSCC patients, we raised a possibility that the microlocalization of CD68(+) TAMs was an indispensable factor for the advance of OSCC.
Assuntos
Antígenos CD/sangue , Antígenos de Diferenciação Mielomonocítica/sangue , Biomarcadores Tumorais/sangue , Carcinoma de Células Escamosas/sangue , Neoplasias Bucais/sangue , Adulto , Idoso , Carcinoma de Células Escamosas/patologia , Intervalo Livre de Doença , Feminino , Humanos , Metástase Linfática , Macrófagos/patologia , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Prognóstico , Modelos de Riscos Proporcionais , Células Estromais/patologia , Resultado do TratamentoRESUMO
The conversion from triene- to diene-typed ansamycins is clarified step by step in Streptomyces seoulensis IFB-A01. Such an intertype convertibility is adopted to establish for the first time the simultaneous mutasynthesis of both types of C17-benzene ansamycins (C17BAs). Three of the newly generated unnatural compounds showed potent cytotoxicity.
Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Rifabutina/análogos & derivados , Rifabutina/síntese química , Streptomyces/química , Antineoplásicos/química , Ensaios de Seleção de Medicamentos Antitumorais , Células Hep G2 , Humanos , Lactamas Macrocíclicas/química , Células MCF-7 , Estrutura Molecular , Rifabutina/química , Rifabutina/farmacologiaRESUMO
Although interleukin (IL) 17A can promote angiogenesis in several tumors, there are limited clinical evidences on cancer about the correlation between serum vascular endothelial growth factor (VEGF) and IL-17F, which is the most homologous to IL-17A. In this study, serum concentration of IL-17F and VEGF from healthy (n = 28), leukoplakia (n = 15), and oral squamous cell carcinoma (OSCC) groups (n = 85) were assessed and showed that IL-17F level was remarkably downregulated from healthy group (394.3 pg/ml) to OSCC group (82.96 pg/ml). Conversely, the OSCC group had a highest level of VEGF (P < 0.05) in whole groups, and there was a negative correlation between IL-17F and VEGF in serum or in the peripheral blood mononuclear cells (PBMCs) at mRNA level. Moreover, the lowest ratio of IL-17F/VEGF was found in OSCC patients (P < 0.05) and lower ratio of IL-17F/VEGF correlated to higher tumor stage and lymph node metastasis. Furthermore, the serum level of IL-17F and the ratio of IL-17F/VEGF were positively associated with the numbers of CD3(+) CD4(+) T cells, which indicated that serum IL-17F could originate from PBMCs during the development of OSCC, and could be used for the diagnosis by effectively distinguishing OSCC patients from healthy individuals.
Assuntos
Carcinoma de Células Escamosas/sangue , Interleucina-17/sangue , Neoplasias Bucais/sangue , Fator A de Crescimento do Endotélio Vascular/sangue , Adulto , Idoso , Biomarcadores Tumorais/sangue , Linfócitos T CD4-Positivos/metabolismo , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patologia , Feminino , Humanos , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/diagnóstico , Neoplasias Bucais/patologia , Neovascularização Patológica/sangue , Neovascularização Patológica/diagnóstico , Neovascularização Patológica/patologia , PrognósticoRESUMO
The endophytic fungus Guignardia mangiferae isolated from Ilex cornuta leaves was shown to produce a family of meroterpenes with toll-like receptor 3 regulating activity (1-9), of which 1-3 possessed new structures. The absolute stereochemistry of 1-3 was assigned through a combination of nuclear magnetic resonance experiments, chemical derivation, CD spectra, and single-crystal X-ray diffraction analyses (CuK α ). The precursor labeled cultivation suggests that these meroterpenes are most likely assembled through terpenoid-shikimate pathways. Moreover, meroterpenes 1-3, 5-7, and 9 selectively upregulate, but 4 and 8 downregulate the toll-like receptor 3 expression in mouse dendritic cells at 10.0 µM.
Assuntos
Ascomicetos/química , Ilex/microbiologia , Terpenos/farmacologia , Receptor 3 Toll-Like/metabolismo , Endófitos , Regulação da Expressão Gênica , Estrutura Molecular , Folhas de Planta/microbiologia , Terpenos/química , Terpenos/isolamento & purificação , Receptor 3 Toll-Like/químicaRESUMO
PURPOSE: To investigate the distribution and clinical outcomes of tumor-infiltrating CD208(+) mature dendritic cells (mDCs) in patients with oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: Using immunohistochemical analysis, the distribution of CD208(+) mDCs in adjacent non-neoplastic tissues (NTs) and tumor tissue, including the tumor stroma (TS) and tumor nest (TN), of 79 patients with OSCC was evaluated. The analysis was quantitative, and the number of positive cells was counted in 5 microscopic high-power fields (×400). At the gene expression level, CD208 expression also was assessed by quantitative polymerase chain reaction. Kaplan Meier analyses were used to analyze the prognostic value of tumor-infiltrating CD208(+) mDCs. RESULTS: The number of tumor-infiltrating CD208(+) mDCs was larger in the TS and TN than in the NTs (P < .0001) and the number of CD208(+) mDCs in the TS was larger in patients with OSCC and lymph node positivity (P < .05). At the gene expression level, CD208 also was upregulated in patients with lymph node positivity (P < .05). The number of infiltrating CD208(+) mDCs was not associated with a patient's survival time. CONCLUSIONS: The number of tumor-infiltrating CD208(+) mDCs in the TS was larger in patients with OSCC and lymph node positivity, but the accumulation of CD208(+) mDCs did not correlate with survival time in patients with OSCC.
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Carcinoma de Células Escamosas/patologia , Células Dendríticas/imunologia , Proteínas de Membrana Lisossomal/imunologia , Neoplasias Bucais/patologia , Proteínas de Neoplasias/imunologia , Análise de Sobrevida , Carcinoma de Células Escamosas/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/imunologia , Reação em Cadeia da PolimeraseRESUMO
Monocyte chemotactic protein-1 (MCP-1/CCL2) and macrophage inflammatory protein-1α (MIP-1α/CCL3) are small chemotactic proteins that have been found in several kinds of tumor tissue samples and function as key regulators of cancer progression. However, the expression of CCL2 and CCL3 in serum samples of oral squamous cell carcinoma (OSCC) patients remains unknown. This study aimed to investigate the prognostic meaning of serum CCL2 and CCL3 in OSCC. The concentration of CCL2 and CCL3 was assessed by ELISA in serum of OSCC patients (n = 98), leukoplakia patients (n = 14), and healthy donors (n = 27). The results showed that the concentration of CCL2 in the OSCC group was significantly lower compared to that in the healthy controls (67.81 vs. 108.1 pg/ml, P < 0.0001). The CCL3 concentration was higher in leukoplakia patients than in OSCC patients and healthy donors (201.9 vs. 153.9 or 118.3 pg/ml, P < 0.05). No significant difference in CCL3 concentration was observed between OSCC patients and healthy donors. However, the OSCC group clearly presented two subclusters, i.e., CCL3 (LOW) and CCL3 (HIGH) OSCC subclusters, in which the serum level of CCL3 was positively related to the tumor size. Interestingly, the ratio of CCL2/CCL3 in OSCC patients was correlated to TNM (tumor, node, metastasis), smoking habits, and differentiation. The receiver operating characteristic (ROC) curve suggests that serum CCL2 is a good diagnostic marker to discriminate OSCC patients from healthy people (cutoff value, 101.1 pg/ml) and the ratio of CCL2/CCL3 also is a good diagnostic marker to discriminate leukoplakia patients and CCL3 (HIGH) OSCC patients from healthy people (cutoff values, 1.080 and 0.424, respectively). These results indicate that CCL2 and CCL3 are associated with progression of OSCC and may be potential biomarkers.
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Biomarcadores Tumorais/sangue , Carcinoma de Células Escamosas/diagnóstico , Quimiocina CCL2/sangue , Quimiocina CCL3/sangue , Neoplasias Bucais/diagnóstico , Adulto , Idoso , Carcinoma de Células Escamosas/sangue , Progressão da Doença , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/sangueRESUMO
OBJECTIVE: We sought to investigate the role and diagnostic value of microRNA 155 (miR-155) in OSCC patients. STUDY DESIGN: Using real-time quantitative polymerase chain reaction analysis, miR-155 expression levels were assessed in OSCC cell lines and a cancerous HB cell line. The correlation between miR-155 expression level and clinical parameters was analyzed in 46 patients with OSCC. In addition, the effects of miR-155 on OSCC cell proliferation were evaluated by modulating its expression using an miR-155 mimic and antisense miR-155. RESULTS: Significant upregulation of miR-155 was found in OSCC cell lines and in tissues of patients with OSCC. The receiver operator characteristic analysis indicated fair-to-good predictability. Overexpression of miR-155 correlated with the histologic grade (P = .033), and the upregulation of miR-155 enhanced OSCC cell proliferation. CONCLUSIONS: In OSSC, upregulation of miR-155 correlated with the histologic grade and can be used as a potential prognostic biomarker.
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Carcinoma de Células Escamosas/metabolismo , MicroRNAs/metabolismo , Neoplasias Bucais/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/patologia , Gradação de Tumores , Estadiamento de Neoplasias , Valor Preditivo dos Testes , Reação em Cadeia da Polimerase em Tempo Real , Células Tumorais Cultivadas , Regulação para CimaRESUMO
BACKGROUND: Long non-coding RNAs play an important role in tumorigenesis, hence, identification of cancer-associated lncRNAs and investigation of their biological functions and molecular mechanisms are important for understanding the development and progression of cancer. Recently, the downregulation of lncRNA MEG3 has been observed in various human cancers. However, its role in non-small cell lung cancer (NSCLC) is unknown. The aim of this study was to examine the expression pattern of MEG3 in NSCLC and to evaluate its biological role and clinical significance in tumor progression. METHODS: Expression of MEG3 was analyzed in 44 NSCLC tissues and 7 NSCLC cell lines by qRT-PCR. Over-expression approaches were used to investigate the biological functions of MEG3 in NSCLC cells. Bisulfite sequencing was used to investigate DNA methylation on MEG3 expression. The effect of MEG3 on proliferation was evaluated by MTT and colony formation assays, and cell apoptosis was evaluated by Hoechst staining and Flow-cytometric analysis. NSCLC cells transfected with pCDNA-MEG3 were injection into nude mice to study the effect of MEG3 on tumorigenesis in vivo . Protein levels of MEG3 targets were determined by western blot analysis. Differences between groups were tested for significance using Student's t-test (two-tailed). RESULTS: MEG3 expression was decreased in non-small cell lung cancer (NSCLC) tumor tissues compared with normal tissues, and associated with advanced pathologic stage, and tumor size. Moreover, patients with lower levels of MEG3 expression had a relatively poor prognosis. Overexpression of MEG3 decreased NSCLC cells proliferation and induced apoptosis in vitro and impeded tumorigenesis in vivo. MDM2 and p53 protein levels were affected by MEG3 over-expression in vitro. CONCLUSIONS: Our findings indicate that MEG3 is significantly down-regulated in NSCLC tissues that could be affected by DNA methylation, and regulates NSCLC cell proliferation and apoptosis, partially via the activition of p53. Thus, MEG3 may represent a new marker of poor prognosis and is a potential therapeutic target for NSCLC intervention.
Assuntos
Apoptose/genética , Carcinoma Pulmonar de Células não Pequenas/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Pulmonares/genética , RNA Longo não Codificante/genética , Proteína Supressora de Tumor p53/genética , Animais , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Transformação Celular Neoplásica/genética , Metilação de DNA , Modelos Animais de Doenças , Feminino , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Metástase Linfática , Masculino , Camundongos , Gradação de Tumores , Estadiamento de Neoplasias , RNA Longo não Codificante/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
Vascular endothelial growth factor (VEGF) is a tumor angiogenesis factor that is important in immune regulation. In our previous study, we found that VEGF expression in the peripheral blood and neoplasm nest from patients with oral squamous cell carcinoma (OSCC) was positively correlated with the course of disease, while an inverse correlation between VEGF expression and dendritic cells (DCs) was identified in the peripheral blood. Therefore, in the present study, we investigated whether inhibition of human VEGF in the human tongue carcinoma cell line Tca8113 had effects on the activity of monocyte-derived DCs. We knocked down the expression of human VEGF in Tca8113 cells using the small interfering RNA (siRNA) technique. Tca8113 cells pre-transfected with siRNA targeting VEGF were co-cultured with monocytederived immature and mature DCs. Cell proliferation was evaluated by a WST-8 assay. Cell apoptosis, cell cycle and cell phenotypes were determined by flow cytometry. The data revealed that downregulation of the human VEGF significantly inhibited the proliferation of Tca8113 cells and increased apoptosis. Inhibition of human VEGF arrested the cell cycle of Tca8113 cells at the G0/G1 phase. Our results showed that the co-culture of DCs with Tca8113 cells markedly inhibited the expression of the mature markers of DCs including HLA-DR, CD80, CD86, CD40 and CD1a, as well as the immature marker CD83, while inhibition of human VEGF in Tca8113 cells significantly reversed these effects. Therefore, human VEGF in Tca8113 cells may not only regulate the cell proliferation and apoptosis of oral squamous cell carcinoma cells, but may also inhibit DC maturation.
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Bioactive natural products offer multiple opportunities for the discovery of novel chemical entities with potential pharmaceutical, nutraceutical and agrochemical applications. Many new organic compounds with novel scaffolds are isolated in small quantities and established methods often fail to determine the structure and bioactivity of such novel natural products. To meet this challenge, we present here a new methodology combining RDC (residual dipolar coupling)-based NMR spectroscopy in microtubes, with a motif-inspired biological assessment strategy. Using only one milligram (ca. 1.5 µmol) of sample, the new protocol established the bioactivity as well as the relative and absolute configuration of vatiparol obtained from Vatica parvifolia. Vatiparol is unique in its unprecedented carbon skeleton and selective inhibitory effect on the expression of monocyte chemo-attractant protein-1 (MCP-1, also known as CCL2). The plausible biosynthetic pathway of vatiparol is briefly discussed. The approach introduced here promises to be widely applicable to the determination of the structure and bioactivity of structurally unknown organic samples available in very limited amounts.
Assuntos
Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Produtos Biológicos/química , Produtos Biológicos/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/química , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Polifenóis/química , Polifenóis/farmacologia , Espectroscopia de Ressonância Magnética/métodos , Modelos Químicos , EstereoisomerismoRESUMO
Cytokine-induced killer (CIK) cells are immune effector cells characterized by co-expression of CD3 and CD56 molecules. We examined the quantities of CIK cells and the changes of these cell expressing NK cell receptors in HIV-1-positive children infected via mother-to-child transmission. The percentage of CIK cells was quantified and the changes in the surface cell receptor profiles in 18 HIV-1-infected children were examined. We found that CIK cell percentages were dramatically increased in HIV-1-infected children. Furthermore, the expressions of CD16, NKp30, NKp44, NKp46, NKp80 and CD244 on CIK cells were decreased, while the expressions of KIR3DL1 and NKG2D on CIK cells were increased in HIV-1-infected children. However, the expressions of KIR2D and NTB-A on CIK cells did not change in the HIV-1-infected children. CIK cells possessed the characteristics of promoting the maturation of dendritic cells and killing functions in HIV-1-infected children. Moreover, serum concentrations of IL-4 and IFN-γ were significantly increased in HIV-1-infected children compared with the HIV-negative controls. These changes likely occurred as a protective mechanism against transmission of maternal HIV-1 virus and thereby helped to limit viral spread, eliminate infected cells and help HIV-1-infected patients to slow the progression to AIDS.
Assuntos
Complexo CD3/imunologia , Antígeno CD56/imunologia , Células Matadoras Induzidas por Citocinas/imunologia , Infecções por HIV/imunologia , HIV-1 , Ativação Linfocitária/imunologia , Povo Asiático , Complexo CD3/biossíntese , Antígeno CD56/biossíntese , Criança , Pré-Escolar , Técnicas de Cocultura , Células Matadoras Induzidas por Citocinas/citologia , Citocinas/análise , Citocinas/biossíntese , Células Dendríticas/citologia , Células Dendríticas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Citometria de Fluxo , Humanos , Transmissão Vertical de Doenças Infecciosas , Masculino , Receptores de Células Matadoras Naturais/biossíntese , Receptores de Células Matadoras Naturais/imunologiaRESUMO
BACKGROUND: Stanniocalcin-1(STC-1) is up-regulated in several cancers including gastric cancer. Evidences suggest that STC-1 is associated with carcinogenesis and angiogenic process. However, it is unclear on the exact role for STC-1 in inducing angiogenesis and tumorigeneisis. METHOD: BGC/STC cells (high-expression of STC-1) and BGC/shSTC cells (low- expression of STC-1) were constructed to investigate the effect of STC-1 on the xenograft tumor growth and angiogenesis in vitro and in vivo. ELISA assay was used to detect the expression of vascular endothelial growth factor (VEGF) in the supernatants. Neutralizing antibody was used to inhibit VEGF expression in supernatants. The expression of phosphorylated -PKCßII, phosphorylated -ERK1/2 and phosphorylated -P38 in the BGC treated with STC-1protein was detected by western blot. RESULTS: STC-1 could promote angiogenesis in vitro and in vivo, and the angiogenesis was consistent with VEGF expression in vitro. Inhibition of VEGF expression in supernatants with neutralizing antibody markedly abolished angiogenesis induced by STC-1 in vitro. The process of STC-1-regulated VEGF expression was mediated via PKCßII and ERK1/2. CONCLUSIONS: STC-1 promotes the expression of VEGF depended on the activation of PKCßII and ERK1/2 pathways. VEGF subsequently enhances tumor angiogenesis which in turn promotes the gastric tumor growth.
Assuntos
Indutores da Angiogênese/metabolismo , Células Endoteliais/metabolismo , Glicoproteínas/metabolismo , Neovascularização Patológica/metabolismo , Neoplasias Gástricas/irrigação sanguínea , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/metabolismo , Indutores da Angiogênese/farmacologia , Western Blotting , Ensaio de Imunoadsorção Enzimática , Glicoproteínas/genética , Humanos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Neovascularização Patológica/patologia , Proteína Quinase C/metabolismo , Proteína Quinase C beta , Transdução de Sinais , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Células Tumorais Cultivadas , Fator A de Crescimento do Endotélio Vascular/farmacologiaRESUMO
Dendritic cells (DCs) play a pivotal role in the pathogenesis of human immunodeficiency virus-1 (HIV-1). Reduced numbers of blood DCs have been observed in individuals with chronic HIV-1 infection. In the present study, we analyzed the expression levels of monocytes, myeloid dendritic cell (mDC) precursors, mDCs, and plasmacytoid dendritic cells (pDCs), in HIV-1-infected patients in China who were infected via different routes of transmission, including heterosexual and homosexual sexual contact, and blood transmission through importation of blood or blood products, to further elucidate their role in HIV. Compared with HIV-negative individuals (n = 40), relative levels of CD11c+CD14â»mDCs, CD11c++CD123(low) mDCs, and CD11câ»CD123+ pDCs in total peripheral blood mononuclear cells (PBMCs) were significantly lower in all HIV patients (n = 93), and in those with blood transmission (n = 26) and heterosexual transmission (n = 43), while relative levels of CD11c+CD14â»mDCs were significantly lower in HIV patients infected via homosexual transmission (n = 24). The results of correlation analysis demonstrated a significant negative correlation between CD4+ T-cell counts and the relative levels of CD11c++CD123(low) mDCs in HIV-I patients infected via blood transmission. There was no significant correlation between CD4+ T-cell counts and the expression level of other DC subpopulations in PBMCs from HIV patients. The results of this study suggest that HIV-1 patients with different routes of transmission exhibit altered expression levels of blood DC subpopulations, which contributes to dysregulated immune responses and pathogenesis of HIV-1.
Assuntos
Células Dendríticas/imunologia , Infecções por HIV/imunologia , HIV-1/imunologia , Adulto , Povo Asiático , Sangue/imunologia , Antígeno CD11c/análise , Contagem de Linfócito CD4 , China , Células Dendríticas/química , Feminino , Humanos , Subunidade alfa de Receptor de Interleucina-3/análise , Receptores de Lipopolissacarídeos/análise , Masculino , Pessoa de Meia-IdadeRESUMO
Natural killer (NK) cells are believed to play a role in the progression of human immunodeficiency virus 1 (HIV-1) disease, and NK cell levels are reduced in individuals with chronic HIV-1 infection. To assess the effects on quantity of NK cells and the changes of NK cell receptors in HIV-1 infected children via mother-to-child transmission, the percentage of NK cells is quantified and the changes in the NK cell receptor profiles in 20 HIV-1 infected children who are not progressing into AIDS were examined. The results showed that NK cell percentage was decreased in the HIV-1 infected children. The expression of NKp30 on NK cells was increased, while the expressions of CD16, NKp44, NKp46, NKp80, NTB-A, CD244, KIR2D, KIR3DL1 and NKG2D on NK cells were decreased in the HIV-1 infected children. NK cell cytolytic activity was elevated in HIV-1 infected children. These results indicate that the acute changes in NK cell percentage and NK cell receptors in HIV-1 infected children are different from the HIV-1 infected adult individuals. Moreover, serum concentrations of IL-18 were elevated in HIV-infected children compared to HIV-uninfected controls. These differences probably play a role in protecting against transmission of maternal HIV-1 virus and guiding the therapeutic strategies for HIV-1 infected children.
Assuntos
Infecções por HIV/imunologia , HIV-1/imunologia , Células Matadoras Naturais/imunologia , Receptores de Células Matadoras Naturais/imunologia , Adolescente , Linfócitos T CD4-Positivos/imunologia , Criança , Pré-Escolar , China , Citotoxicidade Imunológica/imunologia , Feminino , Citometria de Fluxo , Infecções por HIV/transmissão , Humanos , Interleucina-12/sangue , Interleucina-18/sangue , Masculino , Carga Viral/imunologiaRESUMO
Natural killer (NK) cells are believed to play a role in human immunodeficiency virus 1 (HIV-1) disease progression, and NK cell levels are reduced in individuals with chronic HIV-1 infection. In the present study, we compared the frequency and phenotype of peripheral blood CD3-CD56+ NK cells in HIV-1 infected patients in China who were infected through different routes of transmission, including heterosexual and homosexual sexual contact, and blood transmission through injection drug use or importation of blood or blood products. The results showed significantly reduced numbers of CD3-CD56+ NK cells with no association with route of transmission. The expression of CD16 on CD3-CD56+ NK cells in HIV-1 infected patients was similar to that in healthy controls. Among the examined receptor (KIR3DL1, NKp80, NKp44, CD244, NKG2D, and NTBA) expressions, only KIR3DL1 and NKp80 expressions on CD3-CD56+ NK cells were suppressed in HIV-1-infected patients compared to healthy controls, and no significant difference was observed between patients upon comparison of different routes of transmission. A subset of CD3(dim)/CD56+ cells was dramatically increased in HIV-1-infected patients. This study suggests that changes in NK cell count and receptors are not related to the route of HIV-1 transmission. A new subset of CD3(dim)/CD56+ cells emerged only in HIV-1-infected patients, and may play a role in limiting viral spread, eliminating infected cells, and slowing the progression from HIV-1 infection to AIDS.