Nasopharyngitis Patients Research Articles

Serum LINC00339 is a promising biomarker for prognosis prediction of nasopharyngeal carcinoma.

Long non-coding RNAs (lncRNAs) in serum were useful and promising biomarkers for diagnostic and prognostic application. Herein, we investigated the serum lncRNA LINC00339 expression and its role in nasopharyngeal carcinoma. In this study, we recruited a cohort of 129 nasopharyngeal carcinoma patients, 68 patients with nasopharyngitis, and 80 healthy controls. Serum LINC00339 levels were measured by reverse transcription-quantitative polymerase chain reaction. Receiver operating characteristic (ROC) and Kaplan-Meier curve analyses were conducted to evaluate th e clinical role of LINC00339. The effects of linc00339 on cellular activities were measured using CCK-8 and Transwell assays. We observed that serum LINC00339 expression was upregulated in nasopharyngeal carcinoma patients and closely associated with tumor node metastasis stage, lymph node metastasis, and overall survival rate. Meanwhile, ROC analysis showed serum LINC00339 had diagnostic value to distinguish nasopharyngeal carcinoma patients from healthy individuals and nasopharyngitis patients. Silencing of LINC00339 could repress cellular behaviors by targeting miR-152. This study clarified that LINC00339 was upregulated in nasopharyngeal carcinoma, and that serum LINC00339 may act as a diagnostic or prognostic marker, and a hopeful therapeutic target for patients with nasopharyngeal carcinoma.

Noninvasive Diagnosis of Nasopharyngeal Carcinoma Based on Phenotypic Profiling of Viral and Tumor Markers on Plasma Extracellular Vesicles.

Nasopharyngeal carcinoma (NPC) is a malignant tumor commonly associated with Epstein-Barr virus (EBV) infection, and its early diagnosis as well as its differentiation from nasopharyngitis (NPG) remains challenging due to the insufficient sensitivity of routine screening methods in clinical practice. To date, circulating extracellular vesicles (EVs, 40-1000 nm) have shown appealing potential in liquid biopsy for cancer diagnosis and prognosis. Herein, nanoflow cytometry (nFCM) capable of single EV analysis was applied to examine the expression of surface proteins with very low copy numbers on individual EVs as small as 40 nm. The particle concentrations of five EV subsets exposing EBV-encoded latent membrane proteins (LMP1 and LMP2A) and tumor markers (PD-L1, EGFR, and EpCAM) in plasma were determined rapidly via single-particle enumeration. We identified a five-marker panel named EVSUM5 (an unweighted sum of the concentration of the five individual EV subsets) that significantly surpassed the traditional VCA-IgA assay in discriminating NPC patients from both healthy donors and NPG patients with accuracies of 96.3 and 83.1%, respectively. Moreover, EVSUM2 (an unweighted sum of virus-specific LMP1- and LMP2A-positive EVs) could achieve the diagnosis of NPG with an accuracy of 82.6%. Collectively, the work presented a rapid, reliable, and noninvasive method as well as two diagnostic markers to help more accurately differentiate NPC from NPG patients and healthy donors in clinical practice.

ALDH1B1 predicts poor survival for locally advanced nasopharyngeal carcinoma patients.

BackgroundDistant metastasis and local recurrence remain the major reasons of treatment failure in locally advanced nasopharyngeal carcinoma (NPC). Therefore, exploring novel biomarkers for prognosis and sensitivity of radiotherapy in locally advanced NPC is crucial. This retrospective study evaluates the expression and prognostic value of aldehyde dehydrogenase 1B1 (ALDH1B1) for locally advanced NPC patients.MethodsSixty-seven locally advanced NPC patients and 22 chronic nasopharyngitis patients between September 2012 to November 2016 at The First Affiliated Hospital of University of South China were enrolled in this study. The expression of ALDH1B1 in tumor tissues were detected by using immunohistochemistry (IHC).ResultsSignificant difference was observed between NPC groups and Pharyngitis tissues groups, and NPC groups has a higher ratio of high ALDH1B1 expression. ALDH1B1 expression were significantly associated with age and radiotherapy response. The Kaplan-Meier analysis indicated that patients with high ALDH1B1 expression had a poor prognosis both in overall survival (OS) and progression-free survival (PFS). Univariate analysis found that age, radiotherapy response and ALDH1B1 expression were correlated with OS. Besides, factors affecting PFS are radiotherapy response and ALDH1B1 expression. Multivariate analysis revealed that radiotherapy response and ALDH1B1 expression were the independent prognostic factors for OS, whereas radiotherapy response was for PFS.ConclusionsThe expression of ALDH1B1 was correlated with age and radiotherapy response. Patients with high ALDH1B1 expression show a poor prognosis both in OS and DFS. ALDH1B1 expression were the independent prognostic factors for OS.

HOXA9 Expression is Associated with Advanced Tumour Stage and Prognosis in Nasopharyngeal Carcinoma.

BackgroundHomeobox A9 (HOXA9), a member of the HOX protein family, plays diverse biological roles in embryonic development and carcinogenesis. The prognostic value of HOXA9 expression in nasopharyngeal carcinoma (NPC) is not well-defined. The present study aimed to analyse NPC tissue HOXA9 expression and determine prognostic significance by investigating the relationship between HOXA9 expression and clinicopathologic features.MethodsBetween January 2010 and December 2014, 252 NPC patients and 30 chronic nasopharyngitis patients (control group) were recruited to participate in the present study. Correlations between HOXA9 expression level and clinicopathologic features (including survival) were analysed.ResultsHigh HOXA9 expression was significantly associated with clinical stage (p < 0.01) and higher T stage (p < 0.01). In univariate analysis, high HOXA9 expression predicted overall survival (OS) (p = 0.011). In multivariate analysis, HOXA9 over-expression independently and significantly predicted poorer PFS (p < 0.01, hazard ratio (HR) = 2.387, 95% CI [0.876, 6.545]) and OS (p < 0.01, HR = 2.486, 95% CI [1.041, 8.926]).ConclusionHigh HOXA9 expression is an independent prognostic factor associated with advanced tumour stage and poorer survival in NPC patients.

Downregulation of miRNA-429 and upregulation of SOX2 were unfavorable to the prognosis of nasopharyngeal carcinoma.

The purpose of this study was to uncover the correlations of expression levels of microRNA-429 (miRNA-429) and SOX2 with clinical parameters and prognosis of nasopharyngeal carcinoma. Nasopharyngeal carcinoma patients (n=95) and nasopharyngitis patients (n=95) in the same period were enrolled. The relative levels of miRNA-429 and SOX2 in nasopharyngeal tissues collected from these patients were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Then, the potential correlation between miRNA-429 and SOX2 was analyzed by Pearson correlation test. Next, the influences of miRNA-429 and SOX2 levels on clinical parameters of nasopharyngeal carcinoma patients were assessed. At last, the factors influencing the prognosis of nasopharyngeal carcinoma were determined using the Cox regression model. It was found that downregulated miRNA-429 and upregulated SOX2 were observed in nasopharyngeal tissues collected from nasopharyngeal carcinoma patients. MiRNA-429 level was negatively correlated with that of SOX2 in nasopharyngeal carcinoma tissues. In addition, miRNA-429 and SOX2 levels were related to age, tumor differentiation, T stage, N stage, and clinical grade of nasopharyngeal carcinoma patients. Moreover, worse prognosis was seen in nasopharyngeal carcinoma patients expressing low level of miRNA-429 or high level of SOX2. Furthermore, Cox regression analysis showed that T3-T4 stage, moderate to high differentiation, and high level of SOX2 were risk factors, while high level of miRNA-429 was the protective factor for nasopharyngeal carcinoma. Downregulation of miRNA-429 and upregulation of SOX2 are unfavorable to the prognosis of nasopharyngeal carcinoma.

In vivo and in vitro study of co-expression of LMP1 and Cripto-1 in nasopharyngeal carcinoma

IntroductionNasopharyngeal carcinoma, an epithelial-derived malignant tumor which because of its anatomical location and atypical early symptoms, when diagnosed invasion and metastasis often have occurred. This requires a better understanding of the development mechanism, identifying diagnostic markers, and developing new treatment strategies. ObjectiveTo study the relationship of LMP1 and Cripto-1 in nasopharyngeal carcinoma. MethodsThe expression of LMP1 and Cripto-1 in specimens obtained from nasopharyngeal carcinoma patients (n=42) and nasopharyngitis patients (n=22) were examined. The expression of LMP1 and Cripto-1 in LMP1-negative and LMP1-positive (CNE1-LMP1) cells were also examined. ResultsThe expression of LMP1 and Cripto-1 was significantly higher in nasopharyngeal carcinoma than in nasopharyngitis (p<0.05). Their expression in nasopharyngeal carcinoma with metastasis were significantly higher than that without metastasis (p<0.05), which was correlated with TNM staging (p<0.05). High Cripto-1 expression and high proliferation rate were seen in CNE1-LMP1 cells. ConclusionsThe expression of LMP1 and Cripto-1 in nasopharyngeal carcinoma is positively related. Their co-expression might contribute to the proliferation and metastasis of nasopharyngeal carcinoma.

Nasopharyngeal brushing: a convenient and feasible sampling method for nucleic acid-based nasopharyngeal carcinoma research

BackgroundTissue specimens for nasopharyngeal carcinoma (NPC) research are scarce because of sampling difficulties. Previous studies have suggested non-invasive nasopharyngeal brushing as an effective sampling method for NPC diagnosis. The present study aimed to evaluate the feasibility of nasopharyngeal brushing in the acquisition of NPC nucleic acids for research.MethodsNasopharyngeal brushing samples were acquired from 24 healthy individuals and 48 NPC patients. Tissues from 48 NPC and 18 nasopharyngitis patients were collected by endoscopic biopsy. The expression levels of tumor suppressor genes (TSGs) and Epstein–Barr virus (EBV)-encoded microRNAs as well as EBV DNA copy number were measured by quantitative polymerase chain reaction in both types of samples.ResultsAmong six TSGs examined, the expression levels of two genes were significantly decreased in nasopharyngeal brushing and tissue samples from NPC patients as compared with those from healthy/nasopharyngitis individuals. Four EBV-encoded microRNAs, mir-bart1-5p, mir-bart5, mir-bart6-5p, and mir-bart17-5p, were significantly up-regulated in both NPC brushing and tissue samples compared with those from healthy/nasopharyngitis controls (P < 0.001). EBV DNA was significantly increased in both nasopharyngeal brushing samples (P < 0.001) and tissue samples (P < 0.001) from NPC patients in comparison with those from healthy controls.ConclusionsNasopharyngeal brushing can obtain sufficient tumoral materials for the analysis of viral nucleic acid, including EBV-encoded microRNAs and EBV DNA. For the detection of TSG expression, nasopharyngeal brushings was feasible but inferior to tissue samples. This study confirms nasopharyngeal brushing as an applicable sampling method that can aid in nucleic acid-based NPC research.

Integrated analysis of microRNA regulatory network in nasopharyngeal carcinoma with deep sequencing.

BackgroundMicroRNAs (miRNAs) have been shown to play a critical role in the development and progression of nasopharyngeal carcinoma (NPC). Although accumulating studies have been performed on the molecular mechanisms of NPC, the miRNA regulatory networks in cancer progression remain largely unknown. Laser capture microdissection (LCM) and deep sequencing are powerful tools that can help us to detect the integrated view of miRNA-target network.MethodsIllumina Hiseq2000 deep sequencing was used to screen differentially expressed miRNAs in laser-microdessected biopsies between 12 NPC and 8 chronic nasopharyngitis patients. The result was validated by real-time PCR on 201 NPC and 25 chronic nasopharyngitis patients. The potential candidate target genes of the miRNAs were predicted using published target prediction softwares (RNAhybrid, TargetScan, Miranda, PITA), and the overlay part was analyzed in Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological process. The miRNA regulatory network analysis was performed using the Ingenuity Pathway Analysis (IPA) software.ResultsEight differentially expressed miRNAs were identified between NPC and chronic nasopharyngitis patients by deep sequencing. Further qRT-PCR assays confirmed 3 down-regulated miRNAs (miR-34c-5p, miR-375 and miR-449c-5p), 4 up-regulated miRNAs (miR-205-5p, miR-92a-3p, miR-193b-3p and miR-27a-5p). Additionally, the low level of miR-34c-5p (miR-34c) was significantly correlated with advanced TNM stage. GO and KEGG enrichment analyses showed that 914 target genes were involved in cell cycle, cytokine secretion and tumor immunology, and so on. IPA revealed that cancer was the top disease associated with those dysregulated miRNAs, and the genes regulated by miR-34c were in the center of miRNA-mRNA regulatory network, including TP53, CCND1, CDK6, MET and BCL2, and the PI3K/AKT/ mTOR signaling was regarded as a significant function pathway in this network.ConclusionOur study presents the current knowledge of miRNA regulatory network in NPC with combination of bioinformatics analysis and literature research. The hypothesis of miR-34c regulatory pathway may be beneficial in guiding further studies on the molecular mechanism of NPC tumorigenesis.Electronic supplementary materialThe online version of this article (doi:10.1186/s13046-016-0292-4) contains supplementary material, which is available to authorized users.

Evaluation of serum granulysin as a potential biomarker for nasopharyngeal carcinoma

BackgroundGranulysin (GNLY) is excreted from cytotoxic T lymphocytes and natural killer cells, and plays an important role in antitumor immunity. However, few studies have estimated serum GNLY concentrations in patients with nasopharyngeal carcinoma (NPC). We evaluated GNLY as a potential biomarker for NPC. MethodsSerum GNLY concentrations were measured in blood samples taken from 98 NPC patients, 56 nasopharyngitis (NPT) patients, and 99 healthy subjects. The clinical relevance of GNLY in NPC was also investigated. We also assessed the association between serum GNLY and serum immunoglobulin A antibodies against the Epstein–Barr virus (EBV) viral capsid antigen (VCA-IgA) and EBV DNA. ResultsSerum GNLY levels were significantly lower in NPC patients and significantly higher in nasopharyngitis patients compared to healthy controls. Thus, serum GNLY performs well as a biomarker for distinguishing between NPC and NPT. The serum GNLY concentration is elevated with corresponding increases in clinical stage and shows a significant correlation with VCA-IgA and EBV DNA concentration. ConclusionsSerum GNLY is closely associated with the clinical characteristics of NPC and may be a potential biomarker for NPC.

Expression of GRIN2A in benign and malignant nasopharyngeal diseases and its clinical significance.

The gene glutamate receptor, ionotropic, N-methyl D-aspartate 2A (GRIN2A) is associated with development and neuron viability, and our previous studies showed it to be substantially methylated in nasopharyngeal carcinoma, indicating a link to this disease. The aim of this work was to investigate GRIN2A expression and its clinical significance in nasopharyngeal carcinoma, in contrast to nasopharyngitis and nasopharyngeal precancerous lesions. Fifty patients with nasopharyngeal carcinoma were selected as study subjects, while 28 chronic nasopharyngitis patients and 22 individuals with nasopharyngeal precancerous lesions were used as controls. Immunohistochemical analysis was used to study GRIN2A protein expression, and its relationship with nasopharyngeal carcinoma clinical stage and histopathological features were assessed. GRIN2A appeared as yellow staining in the cytoplasm or nucleus. It was strongly expressed in the nasopharyngeal epithelial tissues of patients with chronic nasopharyngitis and in nasopharyngeal precancerous lesions, the proportions of GRIN2A-positive cells being 82.1 and 72.7%, respectively. However, it was weakly expressed in nasopharyngeal carcinoma tissues, with 28.0% of cells testing positive (P < 0.001). No significant difference in the expression of GRIN2A was observed between different clinical stages and pathological grades. We conclude that weak GRIN2A expression is a major feature of nasopharyngeal carcinoma.

Detection of Survivin mRNA in nasopharyngeal carcinoma by real-time fluorescence quantitative RT-PCR

To establish the method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in nasopharyngeal carcinoma (NPC) tissues. The total RNA was extracted from NPC cell line CNE-2 and tissues with Trizol and then been transcribed reversely to cDNA, a method of real time fluorescence quantitative RT-PCR for detecting the expression of Survivin mRNA in NPC tissues had been established, in which chronic nasopharyngitis patients’ nasopharynx tissues treated as control group. The expression of Survivin mRNA all could be detected either in CNE-2 cells, NPC tissues or in chronic nasopharyngitis patients’ nasopharynx tissues, and there was higher the expression level of Survivin mRNA in NPC tissues than which in chronic nasopharyngitis patients’ nasopharynx tissues, the difference was significant (P 0.05). There was no relationship between Survivin mRNA expression and age and sex of NPC patients (P > 0.05). Real time fluorescence quantitative RT-PCR is a rapid, effective and high sensitive method for detecting the expression of Survivin mRNA in NPC tissues. The overexpression of Survivin mRNA may play some roles in pathogenesis of NPC.

Epstein-Barr virus DNA m nasopharyngeal biopsies

The Epstein-Barr virus (EBV) has been closely associated with the undifferentiated form of nasopharyngeal carcinoma (NPC), which is particularly common in the high risk area in southeast China. We have examined 37 nasopharyngeal biopsies from patients within this high risk area, including 31 cases of undifferentiated NPC and 6 cases of patients with nasopharyngitis, for the presence of EBV DNA. We found that 26 of 31 biopsies from NPC patients were EBV DNA positive; 3 of the 6 biopsies from patients diagnosed with nasopharyngitis were also EBV DNA positive. Southern blot analysis of the DNAs obtained from the EBV genome positive biopsies, digested with EcoRI, showed that all preparations from the NPC tumors had only one band corresponding to the EcoRI A fragment when a BamHI W fragment was used as a probe. However, one tumor had an additional band with a molecular weight larger than EcoRI A. The presence of this novel band could indicate the integration of viral DNA into host cellular DNA. DNA from the same biopsies were restricted with BamHI and PstI restriction enzymes. The data obtained from these experiments suggest that the EBV genomes in both the NPC tumor biopsies and biopsies from nasopharyngitis patients obtained from an endemic area in South China may be similar to each other and to the B95-8 EBV isolate with respect to the BamHI Y region of the EBV genome. The data also demonstrate that infection of normal nasopharyngeal epithelial cells with EBV takes place in patients with nasopharyngitis.