Nasopharyngeal Tissues Research Articles

BBOX1-AS1 Accelerates Nasopharyngeal Carcinoma Progression by Sponging miR-3940-3p and Enhancing KPNA2 Upregulation.

BackgroundUpregulation of lncRNA BBOX1 antisense RNA 1 (BBOX1-AS1) has been examined in various tumors. However, its role in nasopharyngeal carcinoma (NPC) remains poorly understood.MethodsRT-qPCR was performed to measure the expression of BBOX1-AS1, KPNA2, and miR-3940-3p. In vitro assays were performed to determine the alteration of cell phenotypes in NPC cells upon transfection or co-transfection with sh-BBOX1-AS1, sh-KPNA2, or miR-3940-3p inhibitor. The BBOX1-AS1–miR-3940-3p and miR-3940-3p-KPNA2 interplay was verified via luciferase reporter and RNA pull-down assays.ResultsHigh BBOX1-AS1 levels were detected in the nasopharyngeal carcinoma tissues. BBOX1-AS1 silencing considerably suppressed the proliferative, migratory, and invasive abilities of NPC cells in vitro. Interestingly, BBOX1-AS1 could specifically bind to miR-3940-3 and abrogate the inhibition of KPNA2 induced by miR-3940-3. Additionally, analysis of tissue samples showed that miR-3940-3 was inversely correlated with BBOX1-AS1 and KPNA2.ConclusionOur findings revealed that the BBOX1-AS1/miR-3940-3/KPNA2 axis is pro-oncogenic in NPC progression, uncovering novel insights into targeted therapy for this disorder.

The rhinopharyngeal flap for reconstruction of lower clival and craniovertebral junction defects.

The endoscopic endonasal approach (EEA) to the lower clivus and craniovertebral junction (CVJ) has been traditionally performed via resection of the nasopharyngeal soft tissues. Alternatively, an inferiorly based rhinopharyngeal (RP) flap (RPF) can be dissected to help reconstruct the postoperative defect and separate it from the oropharynx. To date, there is no evidence regarding the viability and potential clinical impact of the RPF. The aim of this study was to assess RPF viability and its impact on clinical outcome. A retrospective cohort of 60 patients who underwent EEA to the lower clivus and CVJ was studied. The RPF was used in 30 patients (RPF group), and the nasopharyngeal soft tissues were resected in 30 patients (control group). Chordoma was the most common surgical indication in both groups (47% in the RPF group vs 63% in the control group, p = 0.313), followed by odontoid pannus (20% in the RPF group vs 10%, p = 0.313). The two groups did not significantly differ in terms of extent of tumor (p = 0.271), intraoperative CSF leak (p = 0.438), and skull base reconstruction techniques other than the RPF (nasoseptal flap, p = 0.301; fascia lata, p = 0.791; inlay graft, p = 0.793; and prophylactic lumbar drain, p = 0.781). Postoperative soft-tissue enhancement covering the lower clivus and CVJ observed on MRI was significantly higher in the RPF group (100% vs 26%, p < 0.001). The RPF group had a significantly lower rate of nasoseptal flap necrosis (3% vs 20%, p = 0.044) and surgical site infection (3% vs 27%, p = 0.026) while having similar rates of postoperative CSF leakage (17% in the RPF group vs 20%, p = 0.739) and meningitis (7% in the RPF group vs 17%, p = 0.424). Oropharyngeal bacterial flora dominated the infections in the control group but not those in the RPF group, suggesting that the RPF acted as a barrier between the nasopharynx and oropharynx. The RPF provides viable vascularized tissue coverage to the lower clivus and CVJ. Its use was associated with decreased rates of nasoseptal flap necrosis and local infection, likely due to separation from the oropharynx.

The Distribution of M2 Macrophage and Treg in Nasopharyngeal Carcinoma Tumor Tissue and the Correlation with TNM Status and Clinical Stage.

Objective:This study aimed to identify the distribution of M2 macrophage and Treg in Nasopharyngeal Carcinoma (NPC) tumor tissue samples. The presence of these two groups of cells was further correlated to clinical stage, tumor size, the lymphatic node involvement, and metastasis. Methods: The total of 50 formalin-fixed paraffin-embedded (FFPE) NPC tissue samples was collected retrospectively (27 samples) and prospectively (23 samples). Samples were FFPE tissue slices. Immunohistochemistry was done on the FFPE tissue slides using anti-CD-163 and anti-FoxP-3 antibodies for M2 macrophage and Treg detection, respectively. The M2 macrophage interpretation was performed by eye-balling method and the score was divided into 0 (negative), 1 (scant), 2 (focal), and 3 (abundant). The average number of Treg FOXP3+ cells in 5 high power fields (HPF) was calculated. The relationship of M2 macrophage and Treg was tested with Spearman’s correlation. The relationship between M2 macrophage and Treg with clinical stage, tumor size, node involvement and metastasis was tested by chi square, with p<0.1. Results:M2 macrophage and Treg were positive correlated (r=0.469, p<0.001). The presence of M2 macrophage and regulatory T cell (Treg) was significantly correlated to tumor size (p= 0.091 for M2 macrophage and p=0.022 for Treg) and clinical stage (p= 0.030 for M2 macrophage and p= 0.002 for Treg), but did not correlate with lymphatic node involvement and metastasis. Conclusions:In Epstein-Barr virus related NPC tumor microenvironment, the presence of M2 macrophage was correlated with Treg, and both types of the cells were correlated with tumor size and clinical stages.

Comparison of longitudinal SARS-CoV-2 nasopharyngeal specimens reveals the transcriptomic COVIDome

Introduction/ObjectiveSARS-Cov-2 is well established to introduce a cytokine-like storm among select individuals that results in multisystem failure and death. Comorbidities, age, oxygen status, and real-time appraisal of inflammatory markers in the blood have been used to risk stratify patients, however, these clinical markers do not comprehensively characterize the at-risk population or disease course. To understand the molecular underpinnings of the primary site of SARS-CoV-2 infection, here, we interrogated the transcriptomic profile of the nasopharyngeal tissue among paired SARS-CoV-2 specimens.Methods/Case ReportWe performed ribosomal depletion RNAseq on 24 primary samples, including 16 paired samples from 8 unique patients who converted between SARS-CoV-2 negative and positive status via clinical diagnostic qRT-PCR. Additional targeted qRT-PCR was performed for ACE2 and TMPRSS2 in an extension sample of 54 paired specimens from 27 unique patients who converted in their SARS-CoV-2 status on the basis of the qRT- PCR test. Differential gene expression, differential correlative expression with ACE2, and correlative expression with viral load was used to identify genes, which were integral to SARS-CoV-2 pathogenesis, so termed the COVIDome. Gene ontologies, pathways, and reactive infiltrate was assessed between specimens and compared with measures of clinical outcome using regression with appropriate correction for multiple hypotheses.Results (if a Case Study enter NA)We observed significant enrichment for ontologies of lymphocyte activation, specifically interferon gamma signaling; (P<1E-20) and platelet activation (P<1E-5). Genes specifically enriched across all three modules included: ADAMDEC1, EPSTI1, GRIP2, IRF7, KLHDC7B, OAS3, OASL, PIK3R4, RSAD2, and XAF1. Using CIBERSORT to approximate immune cell populations from bulk RNA, we observed and enrichment for CD4 immune cells, which was associated with viral status (P<0.01) while high-risk gene signatures were associated with measures of clinical outcome (P<0.05).ConclusionWe characterized the pathogenesis of SARS-CoV-2 in longitudinal nasopharyngeal samples of COVID- 19 patients and related these molecular manifestations with measures of clinical outcome. As proof of principal, our findings suggest additional study in a large, longitudinal extension sample is warranted to validate and assess molecular features of clinical outcome associated with SARS-CoV-2 infection.

Relationship between miR-338-3p and Clinicopathological Parameters, Prognosis, and STAT3 mRNA Expression in Nasopharyngeal Carcinoma.

Objective To investigate the expression of miR-338-3p in nasopharyngeal carcinoma (NPC) and its relationship with STAT3 mRNA expression as well as their relationship with clinical pathological parameters and prognosis of patients. Methods From September 2016 to September 2018, 71 patients with NPC were selected as the NPC group, and 71 samples of NPC tissues were collected during the operation. A total of 23 patients who underwent biopsy due to chronic nasopharyngitis were selected as the control group and 23 nasopharyngeal mucosal tissues were collected. The expressions of miR-338-3p and STAT3 mRNA in nasopharyngeal tissue of two groups were detected by real-time quantitative PCR, and the relationship between the two was analyzed. To collect clinical data of NPC patients and analyze the relationship between the expressions of miR-338-3p and STAT3 in NPC tissues and clinical pathological parameters of the patients, we followed up the patients with nasopharyngeal carcinoma for three years to observe the relationship between miR-338-3p, STAT3, and the prognosis of the patients. Results The relative expression levels of miR-338-3p in nasopharyngeal tissues of the NPC group and the control group were 0.39 ± 0.05 and 1.01 ± 0.09, respectively (P < 0.05). The relative expression levels of STAT3 mRNA in nasopharyngeal tissues of the NPC group and the control group were 3.82 ± 0.21 and 1.04 ± 0.11, respectively (P > 0.05). miR-338-3p was negatively correlated with the relative expression of STAT3 mRNA in nasopharyngeal carcinoma (r = 0.038, P > 0.05). The expression of miR-338-3p was related to the age of the patient, clinical TNM stage, T stage, and distant metastasis (all P < 0.05). STAT3 expression was correlated with clinical TNM stage, T stage, and distant metastasis in our patient (P < 0.05). The expressions of miR-338-3p and STAT3 in nasopharyngeal carcinoma tissues from different gender, histological type, N stage, M stage, and degree of differentiation showed no statistical differences (P > 0.05). The survival rate of the group with low miR-338-3p expression was significantly lower than that of the group with high miR-338-3p expression (P > 0.05). The survival rate of patients with the high STAT3 expression group was significantly lower than that of patients with the low STAT3 expression group (P > 0.05). Conclusion There is a negative correlation between the low expression of miR-338-3p and the high expression of STAT3 in NPC, which are all related to the TNM stage, T stage, and prognosis of the patient.

Comparison between Endoscopic Microdebrider Adenoidectomy and Conventional Curettage Method

Background: Adenoidectomy is a surgical removal of hypertrophic nasopharyngeal lymphoid tissue forming a part of the Waldeyer's ring. Adenoid hypertrophy causes nasal obstruction and airway problems such as snoring, obstructive sleep apnea, recurrent sinusitis, and/or Eustachian tube dysfunction. These complications frequently lead to a need for adenoidectomy. Objective: To compare between cold curettage and endoscopic assisted powered technique as regard operative time, blood loss, postoperative complications. Patients and Methods: This prospective study included 18 patients, both genders, all suffering from hypertrophied adenoid tissue, presented clinically with mouth breathing, snoring, bilateral nasal obstruction and/or discharge and evidenced radiologically with plain X-ray film lateral view to the nasopharynx. All cases presented to ENT Department Zagazag University, during the period from July 2019 to July 2021, seeking for management of their problem. Patients were divided into two groups (9 patients each); Group I: In which patients were undergone conventional adenoidectomy with adenoid curette. Group II: Patients were undergone endoscopic assisted powered adenoidectomy. Results: Conventional curettage method of adenoidectomy group was associated with more operative bleeding than endoscopic assisted powered adenoidectomy group, but there was no significant difference. Nasal packing of conventional curettage method of adenoidectomy group was distributed as following; one patient (11.1%) required mild packing, 3 patients (33.3%) required moderate packing. While in endoscopic assisted powered adenoidectomy group, one (11.1%) required moderate packing. Conclusions: Endoscope assisted powered adenoidectomy needs to be acknowledged as a safe alternative to conventional adenoidectomy.

LACTB2 renders radioresistance by activating PINK1/Parkin-dependent mitophagy in nasopharyngeal carcinoma

Radiotherapy is a standard and conventional treatment strategy for nasopharyngeal carcinoma (NPC); however, radioresistance remains refractory to clinical outcomes. Understanding the molecular mechanism of radioresistance is crucial for advancing the efficacy of radiotherapy and improving the prognosis of NPC. In this study, β-lactamase-like-protein 2 (LACTB2) was identified as a potential biomarker for radioresistance using tandem mass tag proteomic analysis of NPC cells, gene chip analysis of NPC tissues, and differential gene analysis between NPC and normal nasopharyngeal tissues from the Gene Expression Omnibus database GSE68799. Meanwhile, LACTB2 levels were elevated in the serum of patients with NPC after radiotherapy. Inhibiting LACTB2 levels and mitophagy can sensitize NPC cells to ionizing radiation. In NPC cells, LACTB2 was augmented at the transcription and protein levels after radiation rather than nucleus-cytoplasm-mitochondria transposition to activate PTEN-induced kinase 1 (PINK1) and mitophagy. In addition, LACTB2 was first authenticated to co-locate with PINK1 by interacting with its N-terminal domain. Together, our findings indicate that overexpressed LACTB2 provoked PINK1-dependent mitophagy to promote radioresistance and thus might serve as a prognostic biomarker for NPC radiotherapy.

Профілактика частих повторних захворювань носоглотки в дітей із гіперплазією лімфоглоткового кільця

Актуальність. Лімфоїдна тканина носоглотки є одним із місць скупчення та зростання збудників респіраторної патології. Найбільш уразливими в цій ситуації опиняються діти дошкільного віку з гіперплазією лімфоглоткового кільця, у яких відзначається рекурентний перебіг захворювань верхніх відділів респіраторного тракту. Мета дослідження — оптимізація програми терапії гострих риносинуситів у дітей із гіперплазією лімфоглоткового кільця. Матеріали та методи. Під спостереженням перебувало 120 пацієнтів віком від 3 до 6 років, випадково розподілених у 2 групи по 60 дітей у кожну. Режим застосування: по одному впорскуванню в кожну ніздрю два рази на добу. Натрію хлорид 3% застосовувався у вигляді назальних іригацій у дітей першої групи чотири рази на добу. Натрію хлорид 0,9% застосовувався в дітей другої групи за тією ж схемою. Клінічний метод складався з оцінки даних анамнезу, клінічної картини, загальноклінічних методів дослідження. Брався мазок для цитоморфологічного аналізу слизової оболонки носа. Проводилися фарбування препаратів розведеним розчином Гімзи з подальшою мікроскопією, кількісне визначення секреторного sIgA в слині методом імуноферментного аналізу. Результати оброблялися за допомогою критеріїв Манна — Уїтні, Фішера. Результати. Застосування натрію хлориду 3% у вигляді назальних іригацій справляло статистично значущу регенеруючу та сануючу дію на слизову оболонку носа. Використання натрію хлориду 3% на відміну від застосування натрію хлориду 0,9% не супроводжується зниженням рівня секреторного sIgA в слині, а призводить до його деякого підвищення. Висновок. Використання препарату натрію хлориду 3% у вигляді назальних іригацій у програмі профілактики частих повторних захворювань носоглотки в дітей із гіперплазією лімфоглоткового кільця має більш комплексну дію порівняно із застосуванням натрію хлориду 0,9%.

A first case report of nasopharyngeal Mycobacterium abscessus subspecies massiliense infection

BackgroundMycobacterium abscessus subspecies massiliense is a non-tuberculous mycobacteriosis and was subdivided from Mycobacterium abscessus in 2006. This article is the first report on nasopharyngitis caused by Mycobacterium abscessus subspecies massiliense.Case presentationA 45-year-old woman had an 18-month history of recurrent nasopharyngitis and presented with pain in the throat. Mycobacterial tissue culture and polymerase chain reaction testing revealed the presence of Mycobacterium abscessus subspecies massiliense in the nasopharyngeal tissue. This patient underwent surgery, followed by multiple rounds of chemotherapy with oral and intravenous antibiotic agents for 16 weeks. She has had no recurrence during the 56 weeks since treatment.ConclusionIt is difficult to detect the presence of Mycobacterium abscessus subspecies massiliense in a culture from the swabbing sample. The tissue culture from a biopsy specimen is mandatory for the identification of the species. Currently, no definite treatment policy is available and only empirical treatment is applied. This case is an important for the diagnosis and treatment of this bacterial infection on nasopharynx.

The mechanism of miR-410-3p and miR-34c in nasopharyngeal carcinoma development and progression.

To investigate the relationship between miR-410-3p, miR-34c and nasopharyngeal carcinoma development, we detected the expression of miR-410-3p and miR-34c in nasopharyngeal carcinoma tissues and evaluate its clinical value as a molecular marker for predicting the prognosis in patients with nasopharyngeal carcinoma through clinical case study. To identify the role and mechanism of miR-410-3p and miR-34c in nasopharyngeal carcinoma development and progression. The expression of miR-410-3p and miR-34cin 300 cases of nasopharyngeal carcinoma tissues and 30 cases of paired adjacent normal breast tissues was detected by RT-qPCR. The paired t-test was used to compare the differences of miR-410-3pand miR-34c levels between the nasopharyngeal carcinoma and normal groups. The Chi-square test was used to compare the differences between miR-410-3p and miR-34c expression and clinicopathological factors. The Kaplan-Meier survival curve was used to analyze the relationship between miR-410-3p and miR-34c expression and 5-year overall survival (OS). The Cox proportional hazards regression model was used to evaluate the prognostic value. The results were validated by TCGA database. The expression of miR-410-3p was down-regulated in nasopharyngeal carcinoma tissues compared with that of paired normal tissues (P<0.001). The patients with lower miR-410-3p expression have a higher ratio of positive lymph node status (P=0.039) and a poorer 5-year disease-free survival (P=0.001) and 5-year overall survival (P = 0.002). The expression of mi R-34c was down-regulated in nasopharyngeal carcinoma tissues compared with that of paired normal tissues (P<0.001). Up-regulation of the miR-34c inhibited the viability of paired normal tissues (P<0. 01), but there was no significant change in migration of the paired normal tissues. Downregulation of mi R-34c promoted the viability and migration of paired normal tissues (P<0. 05). The expression of miR-410-3p and miR-34c levels are predictors for the OS in patients with nasopharyngeal carcinoma. The expression of miR-410-3p and miR-34c are molecular markers of early nasopharyngeal carcinoma metastasis and an independent prognostic biomarker for patients with nasopharyngeal carcinoma.

Epstein-Barr virus microRNA BART10-3p promotes dedifferentiation and proliferation of nasopharyngeal carcinoma by targeting ALK7.

Non-keratinizing nasopharyngeal carcinoma, the major subtype of nasopharyngeal carcinoma, is characterized by low differentiation and a close relation to Epstein-Barr virus infection, which indicates a link between Epstein-Barr virus oncogenesis and loss of differentiation, and raises our interest in investigating the involvement of Epstein-Barr virus in nasopharyngeal carcinoma dedifferentiation. Our previous study showed abundant expression of an Epstein-Barr virus-encoded microRNA, BART10-3p, in nasopharyngeal carcinoma tissues, but the association between BART10-3p and nasopharyngeal carcinoma differentiation remains unknown. Here, we examined the expression and prognostic value of BART10-3p, and undertook bioinformatics analysis and functional assays to investigate the influence of BART10-3p on nasopharyngeal carcinoma differentiation and proliferation and the underpinning mechanism. Microarray analysis identified BART10-3p as the most significantly upregulated Epstein-Barr virus-encoded microRNA in nasopharyngeal carcinoma tissues and the upregulation was confirmed in two public datasets. The expression of BART10-3p was an independent unfavorable prognosticator in nasopharyngeal carcinoma and its integration with the clinical stage showed improved prognosis predictive performance. Bioinformatics analysis suggested a potential role of BART10-3p in tumor differentiation and progression. Functional assays demonstrated that BART10-3p could promote nasopharyngeal carcinoma cell dedifferentiation, epithelial-mesenchymal transition, and proliferation in vitro, and tumorigenicity in vivo. Mechanistically, BART10-3p directly targeted the 3'UTR of ALK7 and suppressed its expression. Reconstitution of ALK7 rescued BART10-3p-induced malignant phenotypes. Overall, our study demonstrates that BART10-3p promotes dedifferentiation and proliferation of nasopharyngeal carcinoma by targeting ALK7, suggesting a promising therapeutic opportunity to reverse the malignant phenotypes of nasopharyngeal carcinoma.

On-Site, On-Demand 3D-Printed Nasopharyngeal Swabs to Improve the Access of Coronavirus Disease-19 Testing.

Diagnostic testing that facilitates containment, surveillance, and treatment of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), or future respiratory viruses, depends on a sample collection device that efficiently collects nasopharyngeal tissue and that can be manufactured on site when an outbreak or public health emergency is declared by a government. Here two novel stereolithography‐based three‐dimensional (3D)‐printed nasopharyngeal swabs are reported which are made using a biocompatible and sterilizable photoresist. Such swabs are readily manufactured on‐site and on‐demand to ensure availability, if supply chain shortages emerge. Additionally, the 3D‐printed swabs easily adapt to current workflow and testing procedures in hospital clinical laboratories to allow for effortless scaling up of test kits. Finally, the 3D‐printed nasopharyngeal swabs demonstrate concordant SARS‐CoV‐2 testing results between the 3D‐printed swabs and the COPAN commercial swabs, and enable detection of SARS‐CoV‐2 in clinical samples obtained from autopsies.

Expression of ANCR in nasopharyngeal carcinoma patients and its clinical significance.

Anti-differentiation non-coding RNA (ANCR), a long non-coding RNA, is involved in the development, progression and metastasis of various human cancers. However, its clinical significance in nasopharyngeal carcinoma (NPC) still remains unknown. This study aimed to investigate ANCR expression and its clinical significance in NPC.Totally, 96 NPC tissues and 24 non-cancerous nasopharyngeal mucosa tissues were used. The levels of ANCR were determined by qRT-PCR. Relationship of ANCR with patient clinical characteristics, disease-free survival and overall survival (OS) was evaluated.ANCR expression was increased in NPC tissues compared to non-cancerous nasopharyngeal mucosae. ANCR expression was significantly related to lymph node metastasis, clinical stage, and tumor differentiation (P < .05). Kaplan-Meier survival analysis revealed that high level of ANCR expression was significantly associated with poor disease-free survival but not with OS in NPC patients. Univariate analysis showed a significant association between increased ANCR expression and adverse OS (P < .05), but multivariate analysis suggested that ANCR could not be used as an independent prognostic factor for NPC patients.ANCR is involved in the development and progression of NPC, but whether it can be used as an effective therapeutic target for NPC needs further study.

NASOPHARYNGEAL CARCINOMA IN AN ADOLESCENT AFRICAN AMERICAN MAN PRESENTING AS TEMPOROMANDIBULAR JOINT DISORDER: A CASE REPORT WITH MULTIDETECTOR COMPUTED TOMOGRAPHY AND MAGNETIC RESONANCE IMAGING

<h3>BACKGROUND</h3> Nasopharyngeal carcinoma (NPC) has unique demographic characteristics, risk factors, and clinical presentation. Epstein-Barr virus (EBV) infection and exposure to nitrosamines are the main known contributing risk factors for NPC. The highest risk demographic for NPC, however, is young men of Chinese and Southeast Asian descent. In this case report, we discuss a rare case of NPC in an adolescent African American male patient presenting with symptoms of temporomandibular disorder and with no known predisposing risk factors. <h3>CASE REPORT</h3> A 16-year-old African American male patient presented with initial symptoms of pain in the left temporomandibular joint (TMJ) and left angle of the mandible for 2 months. Lateral neck swelling and one episode of epistaxis were also reported. After a clinical exam and ultrasound examination, the diagnoses of temporomandibular disorder and peritonsillar abscess were considered. Antibiotics were prescribed and a month later multidetector computed tomography (CT) was ordered. The CT findings were significant for an infiltrative malignant nasopharyngeal mass with early skull base involvement and cervical lymphadenopathy. Subsequent magnetic resonance findings were significant for invasion of palatal, paraspinal, and masticatory muscles as well as the carotid sheath and V3 encasement with V3 perineural spread. Tissue sampling of the lesion was definitive for EBV-positive squamous cell carcinoma. The definitive diagnosis was NPC with TNM staging T4 N2c M0. Radiation and chemotherapy were then administered to the patient. Magnetic resonance imaging surveillance studies at 2 and 7 months showed continued tumor involution and lymphadenitis improvement with no evidence of progressive disease. <h3>DISCUSSION/CONCLUSIONS</h3> NPC in young African American patients is exceedingly rare. Nasopharyngeal soft tissue masses in young patients can be misinterpreted as hypertrophic adenoid tissue in cone beam computed tomography images. However, in the presence of unexplained symptoms such as TMJ pain, hearing loss, and epistaxis, a malignancy must be considered and investigated. Careful attention should be paid to skull base bony morphology and foramina for possible perineural spread when nasopharyngeal masses are present.

Immune Microenvironment Change and Involvement of Circular RNAs in TIL Cells of Recurrent Nasopharyngeal Carcinoma.

Nasopharyngeal carcinoma is a malignant tumor that is highly prevalent in southern China and the Southeast Asian belt. Recent studies have shown that the T cells play important regulatory roles in tumorigenesis and progression. We test TIL cell of recurrent nasopharyngeal carcinoma and primary nasopharyngeal carcinoma cell. We found that T cell change in recurrent nasopharyngeal carcinoma and primary nasopharyngeal carcinoma cell. Based on GEO database, we selected differently expressed circRNAs in nasopharyngeal carcinoma tissues. qRTPCR show that some circRNAs also highly expressed in TIL cells. In conclusion, immune microenvironment changed in recurrent nasopharyngeal carcinoma. There is involvement of circular RNAs in this progress, with should be researched further.

MicroRNA-203a-3p affects the biological characteristics of nasopharyngeal carcinoma by targeting vascular endothelial growth factor-C.

Nasopharyngeal carcinoma (NPC) has a high incidence in Southeast Asia and China. This study aims to investigate the effect of miR-203a-3p and vascular endothelial growth factor-C (VEGF-C) on nasopharyngeal carcinoma. In this study, we investigate the transfection of miR-203a-3p mimics and the ability of miR-203a-3p to inhibit in nasopharyngeal carcinoma cell line 5-8F and C666-1, the expression levels of protein AKT, p-AKT in VEGF-C and the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signal pathway by plate clone formation experiment, flow cytometer PI staining method, transwell cell experiment, vasculogenic mimicry experiment, and Western blot. The results showed that miR-203a-3p in the nasopharyngeal carcinoma cell line significantly decreased, while VEGF-C in nasopharyngeal carcinoma tissues and cell lines significantly increased. Furthermore, miR-203a-3p inhibited the proliferation of nasopharyngeal carcinoma cells and blocked the cell cycle of nasopharyngeal carcinoma in the G0/G1 phase, reduced the vasculogenic phenomena and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells, and effectively inhibited the growth of nasopharyngeal carcinoma in vivo. The low expression of VEGF-C can inhibit the proliferation, vasculogenic mimicry, and epithelial-mesenchymal transition of nasopharyngeal carcinoma cells. We found that miR-203a-3p was expressed to a low degree in nasopharyngeal carcinoma. Furthermore, miR-203a-3p regulated the PI3K/AKT signal pathway by downregulating the expression of VEGF-C, thereby inhibiting the proliferation, migration, invasion, vasculogenic mimicry, epithelial-mesenchymal transition, and other malignant biological characteristics of nasopharyngeal carcinoma cells.

High-throughput chromosome conformation capture-based analysis of higher-order chromatin structure in nasopharyngeal carcinoma.

BackgroundFirstly, we aimed to compare the differences of higher-order chromatin structure between nasopharyngeal carcinoma (NPC) and normal nasopharyngeal tissues. The second objective was to analyze the specific chromatin interaction site of NPC and the NPC-related genes regulated by this interaction site.MethodsWe included 6 NPC patients and 6 healthy controls to obtain the sequencing results of highest-throughput chromosome conformation capture (Hi-C) technique, followed by further analysis of the specific chromatin interaction sites in NPC.ResultsWe found an abnormal ultra-long distance interaction site on the chromosome 7p in the CNE210 sample, which was caused by a fusion gene SEPT7P2-PSPH. Additionally, a significant interaction site between chromosome 8q and 3p was revealed in the samples CNE25, CNE29, and CNE211, which was the interaction between 1.5 kb downstream of ASAP1 and 0.8 kb upstream of CTNNB1 gene. Further quantitative polymerase chain reaction (qPCR) revealed that ASAP1 and CTNNB1 genes were more highly expressed in CNE25, CNE29, and CNE211 than in the Np group, preliminarily indicating that this interaction site was likely related to the high expression of ASAP1 and CTNNB1 in NPC.ConclusionsThrough Hi-C analysis, we analyzed the specific chromatin interaction sites associated with NPC, and found the chromosomal translocation and chromatin interaction sites associated with NPC based on statistical analysis. This study has certain guiding significance for in-depth study of the mechanism of NPC occurrence and development.

TRIM59 regulates invasion and migration of nasopharyngeal carcinoma cells by targeted modulation of PPM1B

To investigate whether TRIM59 regulates invasion and metastasis of nasopharyngeal carcinoma cells by targeting PPM1B. We analyzed the expression of TRIM59 in nasopharyngeal carcinoma tissues based on data from TCGA database and detected the expressions of TRIM59 and PPM1B in nasopharyngeal carcinoma and adjacent tissues using Western blotting. We also detected the expressions of TRIM59 and PPM1B at both the mRNA and protein levels in nasopharyngeal carcinoma cell lines using RT-PCR and Western blotting. Stable cell lines with TRIM59 overexpression or knockdown were established in HNE1 cells, in which the targeting relationship between TRIM59 and PPM1B was analyzed using Western blotting and a luciferase reporter gene assay. Transwell chamber assay was used to assess changes in the invasion and migration abilities of HNE1 cells with TRIM59 overexpression or knockdown. Analysis based on TCGA database showed that TRIM59 expression was significantly higher in nasopharyngeal carcinoma tissues than in adjacent tissues (P=0.006); the expression of TRIM59 increased (P=0.01) and PPM1B expression decreased significantly (P=0.03) in nasopharyngeal carcinoma tissues. Compared with HNEpC cells, HNE1 cells expressed a significantly higher level of TRIM59 (P=0.04) but a lower level of PPM1B (P=0.01). Luciferase reporter gene assay indicated that PPM1B was a downstream target gene of TRIM59 and its expression was negatively correlated with TRIM59 expression (P=0.01). In HNE1 cells, TRIM59 overexpression significantly promoted cell invasion (P=0.01) and migration (P=0.02) while TRIM59 knockdown obviously suppressed cell invasion (P=0.01) and migration (P=0.01). TRIM59 knockdown with simultaneous PPM1B overexpression more strongly inhibited invasion (P=0.02) and migration (P=0.01) of HNE1 cells as compared with TRIM59 knockdown alone. TRIM59 regulates invasion and migration of nasopharyngeal carcinoma cells through targeted modulation of PPM1B.

SARS-CoV-2: preliminary study of infected human nasopharyngeal tissue by high resolution microscopy

BackgroundThe novel coronavirus SARS-CoV-2 is the etiological agent of COVID-19. This virus has become one of the most dangerous in recent times with a very high rate of transmission. At present, several publications show the typical crown-shape of the novel coronavirus grown in cell cultures. However, an integral ultramicroscopy study done directly from clinical specimens has not been published.MethodsNasopharyngeal swabs were collected from 12 Cuban individuals, six asymptomatic and RT-PCR negative (negative control) and six others from a COVID-19 symptomatic and RT-PCR positive for SARS CoV-2. Samples were treated with an aldehyde solution and processed by scanning electron microscopy (SEM), confocal microscopy (CM) and, atomic force microscopy. Improvement and segmentation of coronavirus images were performed by a novel mathematical image enhancement algorithm.ResultsThe images of the negative control sample showed the characteristic healthy microvilli morphology at the apical region of the nasal epithelial cells. As expected, they do not display virus-like structures. The images of the positive sample showed characteristic coronavirus-like particles and evident destruction of microvilli. In some regions, virions budding through the cell membrane were observed. Microvilli destruction could explain the anosmia reported by some patients. Virus-particles emerging from the cell-surface with a variable size ranging from 80 to 400 nm were observed by SEM. Viral antigen was identified in the apical cells zone by CM.ConclusionsThe integral microscopy study showed that SARS-CoV-2 has a similar image to SARS-CoV. The application of several high-resolution microscopy techniques to nasopharyngeal samples awaits future use.

WNT8B as an Independent Prognostic Marker for Nasopharyngeal Carcinoma.

Background: Members of the Wnt signaling pathway have been shown to play a role in nasopharyngeal carcinoma (NPC) progression. Aim: The purpose of this study was to investigate WNT8B protein expression in NPC patients using tissue microarray (TMA) analysis and to evaluate its correlation with patient survival and clinical parameters. Methods: A total of 82 NPC cases, together with six normal nasopharyngeal tissue samples, were targeted to construct the TMA blocks. The WNT8B protein expression was evaluated by immunohistochemistry and its correlation to the clinicopathological features was investigated. Results: Sixty-two of 82 (75.6%) cases exhibited high WNT8B protein expression while 20/82 (24.4%) cases appeared to have low WNT8B expression. The univariate analysis revealed that systemic metastasis was associated with patient 5-year survival. The multivariate Cox proportional hazard regression analysis showed that WNT8B expression and systemic metastasis were significantly associated with the survival of NPC patients. Furthermore, there was no correlation found between the WNT8B protein expression and other clinicopathological parameters. Conclusion: Our results suggest that the expression of WNT8B is associated with NPC patients’ survival and could serve as an independent prognostic factor for NPC patients.