interleukin-8 and Nasopharyngeal-Neoplasms

Nasopharyngeal carcinoma (NPC) and Hodgkin's disease (HD) are characterized by their association with Epstein-Barr virus (EBV) and the presence of an intense lymphoid stroma, consisting of T lymphocytes and other reactive cells. In both entities, the tumour cells express viral proteins known to provide target epitopes for cytotoxic T-cells (CTLs), yet in vivo, the tumour cells appear to escape CTL recognition. A comparative in situ hybridization study of cytokine and chemokine gene expression in NPC and HD has been undertaken, focusing on cytokines which are known to be inducible by EBV in vitro. Hodgkin and Reed-Sternberg (HRS) cells expressed interleukin (IL)-6, IL-8, and IL-10, and the thymus and activation regulated chemokine (TARC) in 15/22, 0/22, 5/22, and 16/21 cases, respectively. In NPC, the epithelial tumour cells showed expression of IL-6 in 3/43 cases and of IL-8 in 2/40 cases. There was no detectable expression of IL-10 and TARC in these cases. These data confirm that HRS cells frequently express cytokine and chemokine genes and suggest that this may enable HRS cells to modulate the immune response in their microenvironment and to escape CTL detection. In contrast, NPC tumour cells show only rare expression of IL-6 and IL-8 and no detectable expression of IL-10 and TARC. Thus, the results suggest that the mechanisms employed by the EBV-positive tumour cells to escape immune recognition and destruction differ between HD and NPC.

Topics: Carcinoma; Carcinoma, Squamous Cell; Chemokine CCL17; Chemokines, CC; Cytokines; Epstein-Barr Virus Infections; Gene Expression; Hodgkin Disease; Humans; Immunohistochemistry; In Situ Hybridization; Interleukin-10; Interleukin-6; Interleukin-8; Nasopharyngeal Neoplasms; Reed-Sternberg Cells; RNA, Messenger

Predicting tumor recurrence and death in patients with nasopharyngeal carcinoma (NPC) remains to date challenging. We here analyzed the plasmatic secretomes of NPC untreated and relapsing patients, and explored possible correlations with the clinical and pathological features and survival characteristics of the corresponding patient cohorts, with the aim of identifying novel prognostic biomarkers. This study included 27 controls, 45 untreated NPC and 11 relapsed patients. A set of 14 plasma cytokines were analyzed using Millipore multiplex assay. Nitrites were assessed by Griess method. A comparative analysis of each groups' secretome showed upregulation of IL-8, IL-12p70, IL-10 and IP-10 in untreated patients, and of IL-6, IL-10, MCP-1 and IP-10 in relapsing patients. Nitrites significantly correlated with IL-8 during relapse. Secretomes' network analyses revealed prevalence of high correlations between IL8/IL-17A and IFN-γ/IL12p70 in the control group, between TNF-α/IL-8/IL-6, TNF-α/VEGF/IFN-γ and IL-10/MCP-1 in the untreated group, and between IL-8/IL-6/IL-10, TNF-α/IL-8/IL-6, IL12-p70/VEGF/IL-10/IFN-γ, IL-6/IL-10/IFN-γ and IL-8/IP-10 in the relapse group. IL-12p70, IP-10 and MCP-1 levels respectively associated with gender, age and node metastasis respectively. Recurrence-free survival (RFS) analysis showed that patients presenting High IL-8/Low NO immunological scores presented a combined 80% probability of relapse/death after 53 months (combined log-rank test p = 0.0034; individual p = 0.012 and p = 0.016). Multivariate Cox hazard regression analysis revealed that IL-8 (HR = 7.451; 95% CI [2.398-23.152]; p = 0.001) and treatment type (HR = 0.232; 95% CI 0.072-0.749; p = 0.015) were independent prognostic factors. C&RT decision tree analysis showed that High IL-8/Low NO immunological scores predicted treatment failure in 50% cases starting the 36th month of follow-up (AUC = 1) for all of the studied cases and in 57% cases for patients receiving chemotherapy alone (AUC = 1). Altogether, our results showed that NPC development is accompanied with cytokines deregulation to form specific interaction networks at time of diagnosis and relapse, and demonstrate that High IL-8/Low NO signature may constitute a predictor of poor prognosis which may be useful to improve risk stratification and therapy failure management.

Topics: Chemokine CXCL10; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Nitrites; Prognosis; Recurrence; Secretome; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Non-keratinizing nasopharyngeal carcinoma (NPC) is a malignancy with a poor prognosis for relapsing patients and those with metastatic disease. Here, we identify a novel disease mechanism of NPC which may be its Achilles' heel that makes it susceptible to immunotherapy. CD137 is a potent costimulatory receptor on activated T cells, and CD137 agonists strongly enhance anti-tumor immune responses. A negative feedback mechanism prevents overstimulation by transferring CD137 from T cells to CD137 ligand (CD137L)-expressing antigen presenting cells (APC) during cognate interaction, upon which the CD137-CD137L complex is internalized and degraded. We found ectopic expression of CD137 on 42 of 122 (34.4%) NPC cases, and that CD137 is induced by the Epstein-Barr virus latent membrane protein (LMP) 1. CD137 expression enables NPC to hijack the inbuilt negative feedback mechanism to downregulate the costimulatory CD137L on APC, facilitating its escape from immune surveillance. Further, the ectopically expressed CD137 signals into NPC cells via the p38-MAPK pathway, and induces the expression of IL-6, IL-8 and Laminin γ2. As much as ectopic CD137 expression may support the growth and spread of NPC, it may be a target for its immunotherapeutic elimination. Natural killer cells that express a CD137-specific chimeric antigen receptor induce death in CD137

Topics: 4-1BB Ligand; Animals; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Humans; Interleukin-6; Interleukin-8; Laminin; Mice; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Neoplasm Recurrence, Local; Receptors, Chimeric Antigen; Tumor Necrosis Factor Receptor Superfamily, Member 9

Studies are trying to add immunotherapy to gemcitabine and cisplatin (GP) induction chemotherapy, the standard therapy, in nasopharyngeal carcinoma (NPC) patients with locoregionally advanced disease. However, how the immune system responds to GP remains unknown.. We examined the dynamic changes of circulating immune cells and plasma cytokines in NPC patients administered with GP.. After GP administration, immunosuppressive myeloid cells, including CD11b+CD14+ monocytes, CD33+ myeloid cells, CD33+CD11+ myeloid cells, total MDSCs (CD33+CD11+HLA-DR-/low), monocytic MDSCs, and granulocytic MDSCs decreased significantly. The regulatory T cells and B cells, two important suppressive lymphocyte subpopulations, also decreased. On the other hand, the levels of CD3+ T cells, total B cells, central memory CD4+ T cells, and pro-inflammatory cytokines (including Interleukin [IL]-1β, IL-6, IL-2, IL-5, and IL-8) increased significantly after GP administration. Besides, GP chemotherapy did not weaken the cytotoxic activity and proliferative capacity of T cells.. Our results showed the immune modulation effect of GP induction chemotherapy in locoregionally advanced NPC, providing a solid basis for its combination with immunotherapy.

Topics: Cisplatin; Deoxycytidine; Gemcitabine; HLA-DR Antigens; Humans; Induction Chemotherapy; Interleukin-2; Interleukin-5; Interleukin-6; Interleukin-8; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms

Objective To investigate whether tripartite motif containing 59 (TRIM59) influences the biological behavior of nasopharyngeal carcinoma cells by regulating the nuclear factor κB(NF-κB) signaling pathway. Methods TCGA database was used to predict the expression of TRIM59 in nasopharyngeal carcinoma and adjacent tissues. Reverse transcription PCR and Western blot were used to detect the relative expressions of TRIM59 mRNA and protein in nasopharyngeal carcinoma and paracancerous tissues. With human normal nasal mucosal epithelial cells (HNEpCs) as a control, reverse transcription PCR and Western blot analysis were used to detect the relative expression of TRIM59 mRNA and protein in HNE1 and CNE-2Z nasopharyngeal carcinoma cells. Small interference RNA technology was used to down-regulate the level of TRIM59 in HNE1 cells, while a control group, small interference RNA negative control (si-NC) group and TRIM59 small interference RNA (si-TRIM59) group were set up. MTT assay was used to detect the cell proliferation inhibition rate of each transfection group. Transwell

Topics: Cell Line, Tumor; Cell Proliferation; Down-Regulation; Humans; Interleukin-6; Interleukin-8; Intracellular Signaling Peptides and Proteins; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; NF-kappa B; RNA; RNA, Messenger; Signal Transduction; Survivin; Tripartite Motif Proteins; Tumor Necrosis Factor-alpha; Vimentin

The Epstein-Barr virus M81 strain, isolated from a nasopharyngeal carcinoma, induces potent spontaneous virus production in infected B cells. We found that the M81 non-coding Epstein-Barr-encoded RNA EBER2, which carries polymorphisms that are mainly restricted to viruses found in endemic nasopharyngeal carcinomas, markedly stimulated this process. M81 EBER2 increased CXCL8 expression, and this chemokine enhanced spontaneous lytic replication levels in M81-infected B cells. Both events resulted from the endocytosis of extracellular vesicles containing EBER2 that were generated by neighbouring M81-infected B cells, thereby generating a paracrine loop. These effects were strictly dependent on a functional Toll-like receptor 7 (TLR7), a sensor of single-stranded RNA located in the endosome of these cells. These unique properties of M81 EBER2 could be ascribed to its unusually high expression level and to the ability of its single-stranded region to activate TLR7; both of these properties were dependent on M81-specific polymorphisms. Thus, M81 induced chronic inflammation in its target cells and this resulted in increased virus production. These observations provide a mechanistic molecular link between M81 virus replication-a central viral function and a cancer risk factor-and the production of a chemokine involved in inflammation and carcinogenesis.

Topics: B-Lymphocytes; Chemokines; Epstein-Barr Virus Infections; HEK293 Cells; Herpesvirus 4, Human; Host-Pathogen Interactions; Humans; Interleukin-8; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Oncogenic Viruses; RNA, Untranslated; RNA, Viral; Toll-Like Receptor 7; Virus Cultivation; Virus Replication

The incidence rate of nasopharyngeal cancer (nasopharyngeal carcinoma [NPC]) is much higher in Southeast Asia than in western countries. Interleukin-8 (IL-8), a chemokine produced by macrophages, epithelial cells, airway smooth muscle cells, and endothelial cells, is an important immuno-mediator in the development and progression of many types of cancer. Genetic variations in IL-8 have been associated with the risks of NPC and other cancers. In the current study, we evaluated the role of IL-8 in NPC at the levels of DNA, RNA, and protein in a Taiwanese population. First, in a case-control study, 176 NPC patients and 352 cancer-free controls were genotyped, and the associations of IL-8 T - 251A, C + 781T, C + 1633T, and A + 2767T polymorphisms with NPC risk were evaluated. Second, the NPC tissue samples were assessed for their IL-8 mRNA and protein expression by real-time quantitative reverse transcription polymerase chain reaction (PCR) and Western blotting, respectively. Regarding the IL-8 promoter T - 251A, the TA and AA genotypes were associated with significantly decreased risks of NPC compared with the wild-type TT genotype (adjusted odds ratio = 0.61 and 0.52, 95% confidence interval = 0.47-0.93 and 0.37-0.91, P = .0415 and .0289, respectively). The mRNA and protein expression levels for NPC tissues revealed no significant associations among the 20 NPC samples with different genotypes. These findings suggest that IL-8 may play an important role in the carcinogenesis of NPC in Taiwan.

Topics: Asian People; Carcinoma; Case-Control Studies; Female; Gene-Environment Interaction; Genetic Association Studies; Genetic Predisposition to Disease; Humans; Interleukin-8; Male; Middle Aged; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Promoter Regions, Genetic; RNA, Messenger; Taiwan

Nasopharyngeal carcinoma (NPC) has the highest metastasis potential among head and neck cancers. Distant metastasis is the major cause of treatment failure. Recent studies from our laboratory have revealed that IL-8 promotes NPC metastasis via activation of AKT signaling and induction of epithelial-mesenchymal transition (EMT) in the cells. In the present study, we found that IL-8 treatment for NPC cells resulted in an accumulation of DNMT1 protein through activating AKT1 pathway and consequent DNMT1 protein stabilization. Then DNMT1 suppressed E-cadherin expression by increasing the methylation of its promoter region. LY-294002 blocked IL-8-induced p-AKT1 activation resulting in reduction of DNMT1 and increase of E-cadherin expression, whereas forced demethylation using 5-aza-2'-deoxycytidine restored E-cadherin expression. In conclusion, our study, for the first time, shows that the IL-8/AKT1 signaling pathway stabilizes DNMT1 protein, consequently enhancing hypermethylation of E-cadherin promoter regions and downregulating E-cadherin protein level in NPC cells. Upon blockage of the IL-8/AKT pathway and inhibition of DNMT1, E-cadherin expression can be reversed. These data suggest that targeting the IL-8/AKT1 signaling pathway and DNMT1 may provide a potential therapeutic approach for blocking NPC metastasis.

Topics: Cadherins; Carcinoma; Cell Line, Tumor; CpG Islands; DNA (Cytosine-5-)-Methyltransferase 1; DNA (Cytosine-5-)-Methyltransferases; DNA Methylation; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Promoter Regions, Genetic; Proto-Oncogene Proteins c-akt; Signal Transduction

Interleukin-8 (IL-8) is angiogeneic chemokine that plays a potential role in both development and progression of many human malignancies including nasopharyngeal carcinoma (NPC). Epstein- Barr virus (EBV) is recognized to be an important etiologic agent of NPC as the viral gene products are frequently detected in NPC tissue along with the elevation of antibody titre to the viral protein (VCA-p18+ EBNA1) of IgA in the majority of patients. Elevated plasma of Viral Load is regarded as an important marker for the presence of the disease and for the monitoring of disease progression. However, other serum /plasma parameters such as the level of certain interleukins (IL-8 and IL-10) has also been implicated in NPC progression. The study aimed to investigate the correlations between plasma Viral Load and the level of interleukin (IL-8) and Interleukin (IL-10) in relating these parameters to the stages of NPC. In addition of Viral Load (VCA-p18+EBNA1) IgA, Interleukin-8 and Interleukin-10 before and after therapy will be investigated to seek the possible marker for disease progression. A total of 39 NPC patients and 29 healthy control individuals enrolled in this study. Plasma Viral Load was quantified using real-time quantitative PCR. The Level of plasma interleukins both IL-8 and IL-10 were analyzed using ELISA methods. Results indicated there was a significant decrease in viral load was detected in plasma of NPC patients following therapy. Plasma of viral load was shown to be a good prognosticator for disease progression. There were positive correlation between plasma of viral load and IL-8. These non invasive parameters expressed in blood, could be substitutes of viral load using brushing method, which is invasive. In conclusion that: Viral Load, (VCA-p18+EBNA1) IgA and IL-8 levels are promising markers for the presence of NPC and progression of the disease.

Topics: Adult; Biomarkers; Carcinoma; DNA, Viral; Enzyme-Linked Immunosorbent Assay; Epstein-Barr Virus Nuclear Antigens; Female; Herpesvirus 4, Human; Humans; Interleukin-10; Interleukin-8; Male; Middle Aged; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Prognosis; Viral Load

Radioresistance poses a major challenge in nasopharyngeal carcinoma (NPC) treatment, but little is known about how miRNA (miR) regulates this phenomenon. In this study, we investigated the function and mechanism of miR-203 in NPC radioresistance, one of downregulated miRs in the radioresistant NPC cells identified by our previous microarray analysis. We observed that miR-203 was frequently downregulated in the radioresistant NPC tissues compared with radiosensitive NPC tissues, and its decrement significantly correlated with NPC radioresistance and poor patient survival, and was an independent predictor for reduced patient survival. In vitro radioresponse assays showed that miR-203 mimic markedly decreased NPC cell radioresistance. In a mouse model, therapeutic administration of miR-203 agomir dramatically sensitized NPC xenografts to irradiation. Mechanistically, we confirmed that IL8 was a direct target of miR-203, and found that reduced miR-203 promoted NPC cell radioresistance by activating IL8/AKT signaling. Moreover, the levels of IL8 and phospho-AKT were significantly increased in the radioresistant NPC tissues compared with radiosensitive NPC tissues, and negatively associated with miR-203 level. Our data demonstrate that miR-203 is a critical determinant of NPC radioresponse, and its decrement enhances NPC radioresistance through targeting IL8/AKT signaling, highlighting the therapeutic potential of the miR-203/IL8/AKT signaling axis in NPC radiosensitization.

Topics: 3' Untranslated Regions; Animals; Apoptosis; Blotting, Western; Carcinoma; Cell Line, Tumor; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Interleukin-8; Male; Mice, Nude; MicroRNAs; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Phosphorylation; Proto-Oncogene Proteins c-akt; Radiation Tolerance; Radiotherapy; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Survival Analysis; Tumor Burden; Xenograft Model Antitumor Assays

Radioresistance poses a major challenge in nasopharyngeal carcinoma (NPC) treatment, but little is known about how miRNA regulates this phenomenon. In this study, we investigated the function and mechanism of miR-23a in NPC radioresistance, one of downregulated miRNAs in the radioresistant NPC cells identified by our previous microarray analysis. We observed that miR-23a was frequently downregulated in the radioresistant NPC tissues, and its decrement correlated with NPC radioresistance and poor patient survival, and was an independent predictor for reduced patient survival. In vitro radioresponse assays showed that restoration of miR-23a expression markedly increased NPC cell radiosensitivity. In a mouse model, therapeutic administration of miR-23a agomir dramatically sensitized NPC xenografts to irradiation. Mechanistically, we found that reduced miR-23a promoted NPC cell radioresistance by activating IL-8/Stat3 signaling. Moreover, the levels of IL-8 and phospho-Stat3 were increased in the radioresistance NPC tissues, and negatively associated with miR-23a level. Our data demonstrate that miR-23a is a critical determinant of NPC radioresponse and prognostic predictor for NPC patients, and its decrement enhances NPC radioresistance through activating IL-8/Stat3 signaling, highlighting the therapeutic potential of miR-23a/IL-8/Stat3 signaling axis in NPC radiosensitization.

Topics: Animals; Apoptosis; Blotting, Western; Carcinoma; Cell Line, Tumor; Cell Survival; Female; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Kaplan-Meier Estimate; Male; Mice, Nude; MicroRNAs; Middle Aged; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Prognosis; Radiation Tolerance; Radiation, Ionizing; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Signal Transduction; STAT3 Transcription Factor; Xenograft Model Antitumor Assays

Nasopharyngeal carcinoma (NPC) is one of the leading causes of cancer related death in China. One of the reasons is the absence of tumor specific prognostic markers. The aim of this study was to examine the prognostic values of interleukin-8 (IL-8) and matrix metalloproteinase-9 (MMP-9) in NPC patients. A total of 99 consecutive NPC patients and 40 healthy controls were recruited for this study. Serum levels of IL-8 and MMP-9 were evaluated in NPC patients who were followed up for 5 years. The serum levels of IL-8 and MMP-9 in NPC patients were significantly higher than those in healthy controls (IL-8 26.3 [2.9-68.0] ng/ml vs. 20.3 [2.3-38.6] ng/ml, P < 0.001; MMP-9 23.5 [3.6-52.1] ng/ml vs. 17.3 [2.6-36.9] ng/ml, P = 0.002), respectively. The serum levels of IL-8 and MMP-9 were positively correlated with the N classification (IL-8, P = 0.041, and MMP-9, P < 0.001, respectively) and clinical stage (IL-8, P = 0.022, and MMP-9, P < 0.001, respectively) in NPC patients. Analysis using the Kaplan-Meier method indicated that patients with high levels of IL-8 or MMP-9 had significantly shorter overall survival (OS) (IL-8, P = 0.012; MMP-9, P < 0.001) and disease-free survival (DFS) (IL-8, P = 0.021; MMP-9, P = 0.003) time than those with low levels of MMP-9 or IL-8. Univariate and multivariate analysis revealed elevated MMP-9 level was an independent predictor of shorter OS and DFS. Both MMP-9 and IL-8 are involved in NPC progression. MMP-9 in serum may be the clinically useful indicator for prognostic evaluation in NPC patients.

Topics: Adult; Aged; Biomarkers, Tumor; Carcinoma; Case-Control Studies; Disease-Free Survival; Female; Humans; Interleukin-8; Kaplan-Meier Estimate; Male; Matrix Metalloproteinase 9; Middle Aged; Nasopharyngeal Carcinoma; Nasopharyngeal Neoplasms; Neoplasm Staging; Prognosis

Macrophage migration inhibitory factor (MIF) and CXCL8 (also named IL-8) are strongly expressed in the tissues of nasopharyngeal carcinoma (NPC). However, their role in the growth of NPC has not been fully examined. This study aims to evaluate the functions of MIF and CXCL8 on the growth of NPC tumor spheres. The elevated expression of CXCL8 in tumor over normal tissues was confirmed in 37 pairs of biopsies from NPC patients. In the in vitro study, all the poorly differentiated NPC cell lines, including the EBV-positive C666-1, and the EBV-negative CNE-1, CNE-2, SUNE-1, HNE-1 and HONE-1 cells, were found to express CXCL8 and MIF. Therefore, the EBV-positive C666-1 cell was selected to examine for the role of MIF and CXCL8 in the growth of the NPC tumor spheres. Functional study showed that the growth of C666-1 tumor spheres, under the nutrient poor or growth factor supplemented culture conditions, could be inhibited by the CXCL8 specific peptide inhibitor. The growth of the tumor spheres could also be reduced by the CXCR2 specific inhibitor SB225002 or the PI3K/AKT inhibitor LY294002, indicating that the endogenously produced CXCL8 plays an autocrine role in the growth of the tumor spheres. Further mechanistic studies revealed that the gene expression of CXCL8 could be reduced by the MIF specific small interfering RNA (siRNA) or NF-κB inhibitor parthenolide, and the growth of tumor spheres was also reduced after MIF siRNA transfection. Taken together, the present study highlights the role of MIF/CXCL8/CXCR2 axis in the growth of NPC tumor spheres. Chemotherapeutic interference of this signaling pathway may help to control the growth of the NPC tumor.

Topics: Carcinoma; Cell Line, Tumor; Cell Movement; Cell Proliferation; Chromones; Gene Expression; Gene Knockdown Techniques; Humans; Interleukin-8; Intramolecular Oxidoreductases; Macrophage Migration-Inhibitory Factors; Morpholines; Nasopharyngeal Neoplasms; NF-kappa B; Phenylurea Compounds; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Receptors, Interleukin-8B; RNA, Small Interfering; Signal Transduction; Snail Family Transcription Factors; Spheroids, Cellular; Transcription Factors

Nasopharyngeal carcinoma (NPC) has the highest metastatic potential among head and neck cancers. Distant metastasis is the major cause of treatment failure. The role of interleukin-8 (IL-8) in NPC progression remains unknown. Our multivariate survival analyses of 255 patients with NPC revealed that higher IL-8 expression in primary NPC tissue was an independent prognostic factor for overall survival, disease-free survival, and distant metastasis-free survival of the patients. In vitro study revealed that IL-8 was highly expressed in the established high-metastasis NPC clone S18 relative to the low-metastasis cells. Suppression of IL-8 by short-hairpin RNA reduced the expression of IL-8 in S18 cells and subsequently inhibited migration, invasion, and hepatic metastasis of the cells without influencing cellular growth. Overexpression of IL-8 in S26 cells resulted in increased migration, invasion, and metastasis capabilities of the cells without affecting cellular growth. Exogenous IL-8 enhanced the migration and invasion of low-metastasis CNE-2 cells in a dose-dependent manner. An epithelial-mesenchymal transition (EMT) could be induced by IL-8 in various NPC cell lines. The high level of phosphorylated AKT in S18 cells could be suppressed by knocking down IL-8 expression. Further, IL-8-promoted migration and invasion could be abolished by either the application of the phosphoinositide-3-kinase inhibitor LY294002 or the knock down of AKT expression by using small-interfering RNA. In summary, IL-8 serves as an independent prognostic indicator of overall survival, disease-free survival, and metastasis-free survival for patients with NPC. IL-8 promotes NPC metastasis via autocrine and paracrine means, involving activation of AKT signaling and inducing EMT in NPC cells.

Topics: Base Sequence; DNA Primers; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Epithelial-Mesenchymal Transition; Humans; Interleukin-8; Nasopharyngeal Neoplasms; Neoplasm Metastasis; Prognosis; Proto-Oncogene Proteins c-akt; Real-Time Polymerase Chain Reaction; Signal Transduction

Hepatocyte growth factor (HGF) related tumor angiogenesis and prognosis of patients with nasopharyngeal carcinoma (NPC) has not been identified. The expressions of HGF and IL-8, as well as microvessels density were evaluated in 127 NPC biopsies by immunohistochemical staining. The correlation between these parameters and patient's clinicopathological features was analyzed statistically. In vitro, IL-8 concentration was evaluated in exogenous HGF-treated NPC cell lines by ELISA assay. The presence of EBV was also detected in NPC cells by PCR for Bam HI-W fragment. Both 54.3% (69/127) cases of HGF high-expression in tumor cells and 80.3% (102/127) of HGF high-expression in stromal cells were significantly associated with increased microvessels density, advanced clinical stage, lymph node metastasis and high-expression of IL-8. Angiogenesis exhibited in relation to overall survival of NPC patients (P=0.001), and the patients with HGF and IL-8 dual high-expression tumors had a significantly worse prognosis than those with single protein high-expression and dual low expression tumors (P=0.011 and P=0.026, respectively). Exogenous HGF was observed to promote induction of IL-8 in NPC cells without EBV infection. Co-operating with IL-8, HGF might contribute to a poor prognosis of NPC by inducing angiogenesis through both autocrine and paracrine EBV-independent pathways.

Topics: Adult; Aged; Carcinoma; Cell Line, Tumor; Female; Gene Expression Regulation, Neoplastic; Hepatocyte Growth Factor; Humans; Interleukin-8; Male; Middle Aged; Nasopharyngeal Neoplasms; Neovascularization, Pathologic; Prognosis; Treatment Outcome

We aim at the association of macrophage migration inhibitory factor (MIF) with neovascularization and survival of nasopharyngeal carcinoma (NPC), and determine whether MIF is a valuable prognostic predictor for NPC patients.. One hundred and forty one cases of NPC and 25 normal tissues of nasopharynx were collected. The expression of MIF and interleukin 8 (IL-8) was evaluated in tissues microarray by immunostaining. Intratumoral microvessel density (IMD) in relation to immunostainings and clinicopathological factors were analyzed statistically as well as the follow-up data of patients.. High-expression of both MIF (69.5%) and IL-8 (56.0%) were significantly associated with increased microvessels and lymph node metastasis. High-expression of MIF, IL-8 and higher level of IMD were correlated with either patients' overall survival or disease-specific survival in univariate analysis, but only angiogenesis and lymph node status exhibited in relation to survival of patients as independent prognostic factor of NPC by multivariate analysis. In addition, high-expression of MIF and higher level of IMD were closely associated with locoregional failure of NPC patients.. MIF may contribute to lymph node metastasis in NPC by inducing angiogenesis through the way of upregulation of IL-8 expression in an autocrine EBV-independent pathway.

Topics: Adult; Aged; Case-Control Studies; Female; Follow-Up Studies; Humans; Interleukin-8; Intramolecular Oxidoreductases; Lymphatic Metastasis; Macrophage Migration-Inhibitory Factors; Male; Middle Aged; Nasopharyngeal Neoplasms; Nasopharynx; Neoplasm Staging; Neovascularization, Pathologic; Prognosis; Survival Rate; Tissue Array Analysis

The Epstein-Barr virus (EBV)-encoded EBNA1 protein is expressed in all virus-associated tumours, including nasopharyngeal carcinoma (NPC), where it plays an essential role in EBV genome maintenance, replication and transcription. Previous studies suggest that EBNA1 may have additional effects relevant to oncogenesis, including enhancement of cell survival, raising the possibility that EBNA1 may influence cellular gene expression. We have recently demonstrated by gene expression microarray profiling in an NPC cell model that EBNA1 influences the expression of a range of cellular genes, including those involved in transcription, translation and cell signalling. Here, we report for the first time that EBNA1 enhances activity of the AP-1 transcription factor in NPC cells and demonstrate that this is achieved by EBNA1 binding to the promoters of c-Jun and ATF2, enhancing their expression. In addition, we demonstrate elevated expression of the AP-1 targets interleukin 8, vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1alpha in response to EBNA1 expression, which enhances microtubule formation in an in vitro angiogenesis assay. Furthermore, we confirm elevation of VEGF and the phosphorylated isoforms of c-Jun and ATF2 in NPC biopsies. These findings implicate EBNA1 in the angiogenic process and suggest that this viral protein might directly contribute to the development and aggressively metastatic nature of NPC.

Topics: Adenocarcinoma; Cell Line; Cell Line, Tumor; Epstein-Barr Virus Nuclear Antigens; Gene Expression Regulation, Neoplastic; Genome, Viral; Herpesvirus 4, Human; Humans; Immunohistochemistry; Interleukin-8; Kidney; Microtubules; Nasopharyngeal Neoplasms; Neovascularization, Pathologic; Transcription Factor AP-1; Vascular Endothelial Growth Factor A

The cytokine interleukin-8 (IL-8) may play a role in the pathogenesis of nasopharyngeal carcinoma (NPC) through the modulation of tumor immune response or enhanced angiogenesis. Polymorphism of IL-8 gene, which may affect the production level of cytokine, has been inversely associated with a number of cancers. To test this hypothesis, we investigated the relationship of IL-8 gene polymorphisms and NPC in a Chinese population. We analyzed single nucleotide polymorphisms (SNPs) of IL-8 gene -845 T/C, -738 T/A, -353 A/T, -251 A/T and +678 T/C in 280 patients with NPC and 290 age and sex matched controls, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and polymerase chain reaction-sequence specific primers method (PCR-SSP). There were significant differences in the genotype and allele distribution of -251 A/T polymorphism of the IL-8 gene among cases and controls. The -251 AA and AT genotypes were associated with a significantly increased risk of NPC as compared with the -251 TT genotypes (OR=1.820, 95% CI, 1.120-2.959, P=0.015 and OR=1.590, 95% CI, 1.104-2.290, P=0.013, respectively). Haplotype analysis revealed that the homozygosity of the AAT haplotype (defined by SNPs at positions -353, -251 and +678) of IL-8 gene conveys the highest risk for NPC compared with the homozygosity for the TTC haplotype (OR=1.396; 95% CI, 1.064-1.831; P=0.016). The -251 A/T polymorphism of IL-8 and its haplotype are associated with NPC in a Chinese population. Our data suggests that IL-8 gene may play a role in the development of NPC.

Topics: Case-Control Studies; China; DNA Primers; Female; Genetic Predisposition to Disease; Genotype; Haplotypes; Humans; Interleukin-8; Male; Middle Aged; Nasopharyngeal Neoplasms; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide; Reverse Transcriptase Polymerase Chain Reaction

Interleukin-8 (IL-8) is an angiogenic chemokine that plays a potent role in both development and progression of many human malignancies including nasopharyngeal carcinoma (NPC). In the present study, we evaluated the susceptibility and prognostic implications of the (-251) T/A genetic variation in IL-8 in NPC. We used the allele-specific polymerase chain reaction to characterize the variation of the IL-8 promoter region for 160 unrelated Tunisian patients with NPC and 169 healthy control subjects. There was a significant association between the homozygotes IL-8 (-251) AA genotype and nasopharyngeal carcinoma (OR = 2.46; P = 0.004). The presence of the IL-8 (-251) AA genotype was highly associated with elevated NPC risk for male patients. A significant association was demonstrated between the IL-8 (-251) AA genotype and the aggressive forms of NPC as defined by large tumor size, lymph node metastasis, and advanced stages. Moreover, the presence of the IL-8 (-251) AA genotype indicated a significant association with decreased overall survival. Our findings suggest that the IL-8 promoter polymorphism is associated with increased nasopharyngeal carcinoma risk, particularly in males, as well as disease progress, supporting our hypothesis for IL-8 involvement in NPC pathogenesis.

Topics: Adult; Carcinoma; Disease Progression; Female; Genetic Predisposition to Disease; Humans; Interleukin-8; Male; Nasopharyngeal Neoplasms; Neoplasm Invasiveness; Polymorphism, Single Nucleotide; Prognosis; Promoter Regions, Genetic; Risk Factors

To investigate the distribution of genotype and allele frequencies of the genetic polymorphisms of IFN-gamma and IL-8 in patients with nasopharyngeal carcinoma (NPC) and analyze the relationship between the genetic polymorphisms of IFN-gamma and IL-8 and NPC.. A total of 105 NPC patients and 109 healthy people were recruited in this study. The polymorphism of IL-8-251 locus was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The IFN-gamma CA-repeat polymorphism was determined by polymerase chain reaction and polyacrylamide gel electrophoresis with silver staining. The relationship between the polymorphisms of the two loci and NPC was analyzed.. There was no statistically significant difference in IL-8-251(A/T) genotype and allele frequencies between the NPC patients and the healthy people (P < 0.05). The frequency of IFN-gamma 13 times CA repeats in the NPC patients was significantly lower than that of the healthy people (chi2 = 5.878, P = 0.015). A significant difference in the distribution of the genotype of (CA)13+ / (CA)13+, (CA)13+ / (CA)13- and (CA)13- / (CA)13- between the NPC patients and the healthy people was also found (chi2 = 15.181, P = 0.001).. The IFN-gamma 13 times CA-repeat polymorphism is associated with the onset of NPC. But no association between the polymorphism of IL-8-251 (A/T) locus and NPC is evident. The IFN-gamma CA-repeat polymorphism might play an important role in determining the susceptibility to nasopharyngeal carcinoma.

Topics: Carcinoma; Gene Frequency; Genetic Predisposition to Disease; Genotype; Humans; Interferon-gamma; Interleukin-8; Nasopharyngeal Neoplasms; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length

To investigate the expressions of matrix metalloproteinase-9 (MMP-9) and interleukin-8 (IL-8) in nasopharyngeal carcinoma and their association with Epstein-Barr virus latent membrane protein-1 (LMP-1).. The expressions of MMP-9, IL-8 and LMP-1 were immunohistochemical studied in 53 nasopharyngeal carcinoma sections. We statistically analyzed the correlation of these data and also the relationship between the clinical features and the experimental data of these patients.. The positive expression rate of MMP-9, IL-8 and LMP-1 were all 66.04% (35/53) and their average expression score were (33.19 +/- 29.73)%, (33.46 +/- 30.23)%, (35.49 +/- 29.63)% respectively. The expressions of MMP-9 and IL-8 both showed positive correlation with the expression of LMP-1 (r = 0.792, 0.786 respectively). The expressions of MMP-9, IL-8 and LMP-1 showed significant relationship with lymph nodes metastasis (P < 0.05), but no-significant association with age, gender, pathological classification, and clinical stage.. The expressions of MMP-9, IL-8 and LMP-1 were significantly associated with neck lymph nodes metastasis in nasopharyngeal carcinoma. The correlation between LMP-1 and MMP-9, IL-8 showed LMP-1 might enhance neck lymph nodes metastasis by up-regulating the expressions of MMP-9 and IL-8.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Female; Humans; Interleukin-8; Lymphatic Metastasis; Male; Matrix Metalloproteinase 9; Middle Aged; Nasopharyngeal Neoplasms; Neck; Prognosis; Viral Matrix Proteins; Young Adult

Cytokines such as IL-10 and IL-18 seem to be involved in the inflammatory response of undifferentiated carcinoma of nasopharyngeal type (UCNT). The aim of this study was to evaluate the correlation between functional single nucleotide polymorphisms (SNPs) in the promoter region of IL-10 and IL-18 genes and the virological and clinical characteristics in a large case series of Caucasian patients suffering from UCNT, a tumor regularly associated with the Epstein Barr Virus (EBV).. Eighty-nine patients with histologically confirmed UCNT and 130 healthy donors were included in our study. DNA was examined for the polymorphisms of IL-10 gene at positions -1082, -819, -592 by direct sequencing and IL-18 gene at position -607 and -137 by allele-specific PCR. EBV DNA serum viremia was evaluated by QC-PCR.. The distributions of the IL-10 and IL-18 genetic variants were not different between UCNT patients and healthy controls. The frequency of IL-10 -1082G allele, which is associated with high IL-10 expression, showed a nearly statistically significant increase in UCNT patients EBV DNA-negative as compared to healthy controls (OR=3.3 95% CI: 1.2-9.8). Subjects with C/C or C/G combined IL-18 genotypes showed an increased risk of being with Stages III-IV (OR=2.1 95% CI: 1.2-6.6).. This study was performed to improve the definition of the pathogenetic factors implicated in UCNT by addressing the correlation between cytokine polymorphisms and clinical parameters. This is the first study investigating the possible role of the IL-18 and IL-10 polymorphisms in the development and outcome of UCNT. In our genetic analysis there is no evidence for involvement of IL-10 promoter polymorphisms alone in the genetic predisposition to this tumor. On the other hand, IL18 genetic variants may represent a genetic risk factor for tumor aggressiveness.

Topics: Carcinoma; Case-Control Studies; Cohort Studies; Epstein-Barr Virus Infections; Female; Genetic Predisposition to Disease; Genotype; Humans; Inflammation; Interleukin-10; Interleukin-8; Italy; Male; Middle Aged; Nasopharyngeal Neoplasms; Polymerase Chain Reaction; Polymorphism, Single Nucleotide; Prognosis; Promoter Regions, Genetic; Risk Factors

Nasopharyngeal carcinoma (NPC) shows highly invasive and metastatic features. This study aims to investigate macrophage migration inhibitory factor (MIF)-induced invasion of NPC cells in vitro and the effects on matrix metalloproteinases (MMPs) and interleukin-8 (IL-8), and to study the mechanism of tumor cell invasion and metastasis in the early stage of NPC.. Two nasopharyngeal carcinoma cell lines, CNE-1 and CNE-2, were adopted in this study. The NPC cell invasion and migration were evaluated by microinvasion assay. The variation of expression percentages of MMP2- or MMP9-positive cells was detected by flow cytometry in two cell lines with or without MIF treatment. Western blotting and RT-PCR were used to assay the protein and mRNA expressions of MMP2 and MMP9. The IL-8 concentration secreted by NPC cells was compared with the cells with different treatments using ELISA.. After treating with MIF for 48 hours, the cell numbers of CNE-1 and CNE-2 which went through the 8-microm filter membrane were increased. Compared with non-MIF treated NPC cells, significant difference could be found both in CNE-1 (P = 0.005) and CNE-2 cells (P = 0.001). The percentages of MMP9-positive cells were significantly increased in both CNE-1 [from (28.5 +/- 2.5)% to (82.4 +/- 3.5)%, P = 0.001] and CNE-2 [from (32.8 +/- 3.5)% to (86.1 +/- 1.6)%, P = 0.002]. The relative intensity of MMP9 protein expression was also enhanced in both cell lines (CNE-1: from 83.1 +/- 6.0 to 242.9 +/- 22.9, P = 0.002; CNE-2: from 84.4 +/- 4.3 to 278.9 +/- 29.7, P = 0.003). Correspondingly, the increased MMP9 mRNA expression level was significantly detectable in both cell lines. The concentration of IL-8 in the supernatant of CNE-2 was higher [(1201.8 +/- 593.3) pg/ml] after treatment. It was also remarkably higher than that in the supernatant of CNE-2 without treatment (P = 0.026). However, there was no significant difference in the concentration variation of IL-8 in CNE-1 (P = 0.581), while the IL-8 mRNA level was only enhanced in CNE-2.. MIF can induce potent invasion of NPC cell lines in vitro, and the infiltrating lymphocytes in NPC might be responsible for the invasion and metastasis of tumor cells. MIF cytokine which is secreted by these infiltrating lymphocytes might contribute to the invasion as well as metastasis of NPC in the early stages by induction of MMP9 and IL-8 in an indirect pathway.

Topics: Blotting, Western; Cell Line, Tumor; Electrophoresis, Polyacrylamide Gel; Humans; Interleukin-8; Macrophage Migration-Inhibitory Factors; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Nasopharyngeal Neoplasms; Neoplasm Invasiveness; Reverse Transcriptase Polymerase Chain Reaction

Although nasopharyngeal carcinoma (NPC) shows highly invasive and metastatic features than other head and neck carcinomas, the major relevant mechanism is still unknown. This study was designed to investigate whether the macrophage migration inhibitory factor (MIF) can affect the expression of matrix metalloproteinase 2, 9 (MMP-2, MMP-9) and interleukin 8 (IL-8) in nasopharyngeal carcinoma (NPC) cell strains.. Two nasopharyngeal carcinoma cell strains, CNE-1 and CNE-2 were adopted in this study. The variations of expression percentages of MMP-2 or MMP-9-positive cells detected by flow cytometry in NPC cell strains with or without MIF activation were compared. Western blot analysis and reverse transcriptase-polymerase chain reaction (RT-PCR) were applied to evaluate the protein and mRNA expression level of MMP-2 and MMP-9 in cell strains treated with and without MIF, respectively. The concentration of IL-8 in the supernatant of the cells with different treatments was tested using enzyme-linked immunosorbent assay (ELISA).. (1)After treatment with MIF for 24 h, the percentage of MMP-9-positive cells was significantly increased in both CNE-1 (from 28.5+/-2.5% to 82.4+/-3.5%, P=0.001) and CNE-2 (from 32.8+/-3.5% to 86.1+/-1.6%, P=0.002). However, the percentages of MMP-2-positive cells did not significantly change between these two cell strains with or without MIF treatment (P >0.05). (2) The relative intensity of MMP-9 protein expression was also enhanced in both cell strains (CNE-1:from 83.1+/-6.0 to 242.9+/-22.9, P=0.002; CNE-2:from 84.4+/-4.3 to 278.9+/-29.7, P=0.003) and there was no significant difference in MMP-2 expression intensity either in CNE-1 or CNE-2. (3)The IL-8 concentration in CNE-2 supernatant was 1201.8+/-593.3 pg/ml after treatment with MIF for 24 h, remarkably higher than that without treatment (32.7+/-20.1 pg/ml, P=0.026). However, there was no detectable difference of IL-8 concentration found in CNE-1 (P=0.581). (4)The expression level of MMP-9 mRNA, but not of MMP-2 mRNA was significantly increased both in CNE-1 and CNE-2 after treatment with MIF. In addition, the IL-8 mRNA level was only enhanced in CNE-2 but not in CNE-1.. MIF cytokine might play an important role in neoplastic cell invasion and metastasis by up-regulating the expression of MMP-9 and IL-8 in NPC cells through the pathway of activation of their gene transcription.

Topics: Cell Line, Tumor; Humans; Interleukin-8; Macrophage Migration-Inhibitory Factors; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Nasopharyngeal Neoplasms; Neoplasm Invasiveness; RNA, Messenger

To study whether macrophage migration inhibitory factor (MIF) can increase the ability of invasion of nasopharyngeal carcinoma cell lines in vitro, and to investigate the mechanism of invasion and metastasis of tumor cells during the early stage of nasopharyngeal carcinoma (NPC).. The invasion and migration of NPC cell lines, CNE-1 and CNE-2, were evaluated by micron-migration assay in a chamber with 8- micro m porosity polycarbonate filter membrane. Flow cytometry and western blotting were adopted respectively to evaluate the protein expression level of matrix metalloproteinase 2 and 9 (MMP2, MMP9) in MIF treated or non-treated tumor cell lines. The concentrations of interleukin 8 (IL-8) secreted into the culture supernatant by the cells were measured by using Enzyme-linked immunoabsorbent assay (ELISA).. (1) After treatment with MIF for 24 hours, the number of cells passing through the 8- micro m filter membrane were increased in CNE-1 (113.7 +/- 20.9) and CNE-2 (311.3 +/- 48.9), as compared with that of non-MIF treated NPC cells. A significant statistic difference (P = 0.005, P = 0.001) was obtained in both CNE-1 and CNE-2 cells. (2) After treatment with MIF, the number of MMP9-positive cells increased in both CNE-1 (from 28.5% +/- 2.45% to 82.4% +/- 3.49%, P = 0.001) and CNE-2 (from 32.8% +/- 3.48% to 86.1% +/- 1.62%, P = 0.002) cell lines. In addition, an enhanced MMP9 protein expression up to 3-fold was observed in both cell lines. However, the expression level of MMP2 did not changed significantly between treated and non-treated cell lines (P > 0.05). (3) The concentration of IL-8 in the culture supernatant of CNE-2 was 1201.8 +/- 593.3 pg/ml after treatment with MIF for 24 h, remarkably higher than that without MIF treatment (32.7 +/- 20.1 pg/ml, P = 0.026). A similar change was not detected in CNE-1 (P = 0.581) cells.. (1) MIF can increase cell migration of CNE-1 and CNE-2 NPC cell lines in vitro. (2) A higher expression level of MMP9 and an up-regulated IL-8 by MIF may play a very important role in the progress of NPC, such as invasion and metastasis.

Topics: Cell Line, Tumor; Cell Movement; Enzyme-Linked Immunosorbent Assay; Humans; Interleukin-8; Macrophage Migration-Inhibitory Factors; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Nasopharyngeal Neoplasms; Neoplasm Invasiveness

Nasopharyngeal carcinoma (NPC) is a highly invasive and metastatic malignant tumor and is associated with Epstein-Barr virus (EBV) infection that exhibits type II latency. Angiogenesis is essential for tumor growth, invasion, and metastasis. Our previous studies have indicated that interleukin (IL)-8 was over-expressed in many NPC tissues and was found to be significantly correlated with angiogenesis by immunohistochemistry.. In vitro design.. The influence of the EBV genome for IL-8 gene expression was studied using the EBV-genome-positive and -negative epithelial/NPC hybrid cell line NPC-KT. The EBV-positive and -negative clones were selected by polymerase chain reaction and in situ hybridization.. EBV-positive clones expressed abundant IL-8 mRNA compared with EBV-negative clones. This result indicated that over-expression of IL-8 depended on the presence of EBV genomes in NPC-KT cells. Two encoded genes, latent membrane protein (LMP)1 and EBV-encoded small RNAs (EBERs), expressed in NPC were transfected in EBV-negative NPC-KT cells. LMP1 transactivated the IL-8 promoter, whereas EBERs did not. Moreover, the nuclear factor (NF)-kappa B binding site in the IL-8 promoter was essential for the response to LMP1, and the activator protein (AP)-1 binding site played only a partial role.. LMP1 induces IL-8 mainly through the activation of NF-kappa B and partly through AP-1 in NPC model cell lines, NPC-KT, and this suggests that LMP1 plays an important role in the angiogenesis of NPC.

Topics: Blotting, Northern; Carcinoma; Cell Line, Tumor; Cloning, Organism; Epstein-Barr Virus Infections; Humans; Immunohistochemistry; In Situ Hybridization; Interleukin-8; Nasopharyngeal Neoplasms; NF-kappa B; Plasmids; Point Mutation; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Transfection; Viral Matrix Proteins

Interleukin-8 (Il-8) and IL-10 are cytokines associated with the Epstein Barr Virus (EBV), which is linked with nasopharyngeal cancer (NPC). In the present study, we investigated the endogenous production of IL-8 and IL-10 in vitro by two EBV-positive NPC cell lines, viz., HK1 and CNE-2. IL-8 was expressed by both cell lines although the level of IL-8 was 2-fold higher in supernatant from HK1 cells. Incubation with hypericin, a natural photosensitizer increased IL-8 significantly but only in HK1 cells. Hypericin-based photodynamic therapy (PDT) did not alter the expression of IL-8 levels. Il-10 was not constitutively expressed in either cell line and could not be induced by PDT. It is interesting that PDT which is known to upregulate IL-8 transcription via reactive oxygen species and activate the IL-10 promoter did not alter IL-8 levels in either of the NPC cell lines nor induced the production of IL-10.

Topics: Anthracenes; Cytokines; Flow Cytometry; Humans; Interleukin-10; Interleukin-8; Nasopharyngeal Neoplasms; Perylene; Photochemotherapy; Tumor Cells, Cultured

The EBV latent membrane protein-1 (LMP-1) is a multifunctional protein. Recently, the contribution of LMP-1 to the metastasis of nasopharyngeal carcinoma (NPC) has been suggested. Angiogenesis is a key step for metastasis. Thus, the association of LMP-1 to neovascularization of NPC was examined in this study.. The association of LMP-1 to angiogenesis in 39 patients with NPC was evaluated by immunohistochemical study, and then induction of angiogenic factors by LMP-1 was examined by ELISA and luciferase reporter assay.. In an immunohistochemical study, the expression of LMP-1 was significantly correlated to microvessel counts (P = 0.0003), suggesting that LMP-1 may induce some angiogenic factors. Therefore, we studied the relationship between LMP-1 expression and interleukin-8 (IL-8), vascular endothelial growth factor (VEGF), and basic fibroblast growth factor (bFGF) expression by immunohistochemical analysis. IL-8, VEGF, and bFGF expression were correlated to microvessel counts, but only IL-8 expression was significantly correlated to LMP-1 expression (P < 0.0001). Transfection with LMP-1 expression plasmid induced IL-8 protein expression in C33A cells. The expression of LMP-1 transactivated IL-8 promoter, as demonstrated by IL-8 promoter luciferase reporter assay. Mutation of the nuclear factor kappaB responsive element in the IL-8 promoter region completely abolished transactivation by LMP-1, whereas mutation of the activator protein responsive element did not affect promoter activity.. These results suggested that LMP-1 induces expression of IL-8 through the nuclear factor kappaB binding site, which may contribute in part to angiogenesis in NPC.

Topics: Angiogenesis Inducing Agents; Base Sequence; Binding Sites; Blood Vessels; Epstein-Barr Virus Infections; Herpesvirus 4, Human; Humans; Immunohistochemistry; Interleukin-8; Luciferases; Mutation; Nasopharyngeal Neoplasms; Neovascularization, Pathologic; NF-kappa B; Plasmids; Promoter Regions, Genetic; Protein Binding; Recombinant Fusion Proteins; Transcription Factor AP-1; Transcriptional Activation; Tumor Cells, Cultured; Viral Matrix Proteins; von Willebrand Factor

Nasopharyngeal carcinoma (NPC) is a common cancer among southern Chinese. The profile of gene expression in NPC cells is largely unknown. In this study, we have examined differential gene expression in non-malignant and malignant nasopharyngeal epithelial (NPE) cells using a cDNA array hybridization method. A total of 42 genes were identified to be expressed in either non-malignant and malignant NPE cells or both. Thirteen of these genes were overexpressed in malignant NPE cells. These includes nuclear factor (NF90), FOS-related antigen 1 (FRA- 1), cytoplasmic dynein light chain (HDLC1), replication factor C (RFC1), nucleoside diphosphate kinase B, UV excision repair protein (RAD23A), insulin-like growth factor receptor II, transcription initiation factor TFIID subunit (TAFII31), growth factor receptor-bound protein 2 (GRB2), UV excision repair protein (RAD23B), glutathione peroxidase, Y box binding protein 1 and heat shock protein 86. In contrast, expression of nine genes was suppressed in malignant NPE cells. These includes calgranulin A, calgranulin B, neutrophil activating protein (ENA-78), heat shock protein 27, integrin beta-1, integrin beta-4, cyclin-dependent kinase inhibitor 1A (p21), interleukin-8 and tyrosine protein kinase receptor (RET). Differential expression of calgranulin A, calgraunlin B, ENA-78, FRA-1 and NF90 in non-malignant and malignant nasopharyngeal epithelial cells was confirmed by RT-PCR analysis.

Topics: Calcium-Binding Proteins; Calgranulin A; Calgranulin B; Chemokine CXCL5; Chemokines, CXC; DNA-Binding Proteins; DNA, Complementary; Gene Expression Regulation, Neoplastic; Humans; Interleukin-8; Nasopharyngeal Neoplasms; Nasopharynx; NFATC Transcription Factors; Nuclear Factor 90 Proteins; Nuclear Proteins; Proto-Oncogene Proteins c-fos; Reverse Transcriptase Polymerase Chain Reaction; Transcription Factors; Tumor Cells, Cultured

Using polymerase chain reaction (PCR), we confirmed the expression of interleukin-1 alpha (IL-1 alpha) by the human nasopharyngeal carcinoma (NPC) cell line C15 without contribution of either human IL-1 beta or mouse IL-1 alpha in the biological activity previously found in C15. However we showed that IL-1 alpha was not expressed in all NPCs. IL-1 beta and/or tumor necrosis factor (TNF)-alpha genes could also be activated, independently from the number of Epstein Barr Virus (EBV) copies harbored by the cells. Interestingly, the primary tumor C15 showed a profile of TNF-sensitive tumor while C17, C18 and C19 which were derived from metastasis have a typical profile of TNF-resistant cells. Furthermore, the inflammatory cytokines whose genes are classically induced by IL-1 and TNF were found expressed only in C17 and C19 suggesting another level of heterogeneity among NPCs.

Topics: Animals; Base Sequence; Chemokine CCL2; Chemotactic Factors; Cytokines; Gene Expression; Genes, Viral; Herpesvirus 4, Human; Humans; Interleukin-1; Interleukin-8; Mice; Mice, Nude; Molecular Sequence Data; Nasopharyngeal Neoplasms; Polymerase Chain Reaction; Receptors, Cell Surface; Receptors, Tumor Necrosis Factor; RNA, Messenger; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha