CircRNA_0008668 Sponges miR-1301-3p to Promote the Inflammatory Response in Allergic Rhinitis by Upregulating GATA6 Expression.

AIMS: We aim to explore GATA6 modulation in allergic rhinitis (AR). BACKGROUND: Circular RNAs (circRNAs) and microRNAs (miRNAs) are involved in inflammatory responses; GATA6 is also known to regulate multiple inflammatory pathways. However, the mechanism of regulation of AR between them is unclear. OBJECTIVE: We expect that this study will provide new treatment options for AR from a GATA6 perspective. METHODS: In vitro, AR models were employed to examine the efficacy of our study, where we utilized monoclonal anti-2,4,6-dinitrophenyl immunoglobulin (Ig) E/human serum albumin (DNP-IgE/HSA) to induce rat basophilic leukemia cells (RBL-2H3 cells). Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was performed to measure the expression of circ_0008668, miR-1301-3p, GATA6, and cellular inflammatory markers. Enzyme-linked immunosorbent assay (ELISA) was used to measure concentrations of beta-hexosaminidase, histamine, and cellular inflammatory factors including TNF-α, IL-1ß, IL-4, and IL-5. In addition, western blot, RNA pull-down, and luciferase assays were performed to validate the molecular mechanism by which circ_ 0008668 and miR-1301-3p interactions promote GATA6 to ameliorate the inflammatory state of RBL-2H3 cells. RESULTS: In the in vitro model of AR, the expression levels of circ_0008668 and GATA6 were elevated, whereas that of miR-1301-3p was decreased. Pull-down assays confirmed that circ_0008668 efficiently binds miR-1301-3p and its overexpression leads to upregulation of the levels of GATA6, cellular inflammatory factors (IL-4, IL-5, TNF-α, and IL-1ß), and markers associated with inflammatory signaling pathways (NLRP3, ERK1/2, and P65 protein phosphorylation). In addition, miR-inhibitor with circRNA enhanced GATA6 and NLPR3 expression and activated inflammatory pathway activity. In particular, miR-mimic was effective in reversing the onset of this inflammatory state. CONCLUSION: Our results indicate that circ_0008668 promotes the inflammatory state of mast cells by sponging miR-1301-3p to target GATA6, which in turn affects the allergic response to AR. This process could improve the current diagnosis of AR patients and clinical treatment.

Altered circular RNA expression profiles in an ovalbumin-induced murine model of allergic rhinitis

Background Emerging evidence shows that circular RNAs (circRNAs) participate in the pathogenesis of multiple immune diseases. However, few studies have focused on the mechanisms of circRNAs involved in allergic rhinitis (AR). Methods This study performed an RNA sequence (RNA-seq) profiling to identify the expression of circRNAs in nasal mucosa from ovalbumin-induced AR murine models and normal controls. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was then conducted to validate the differential expression of circRNAs. Bioinformatics analysis was applied to demonstrate the biological functions of the dysregulated circRNAs. Results A total of 86 distinct circRNA candidates were sequenced, of which 51 were upregulated and 35 were downregulated. The T cell receptor, B cell receptor, and calcium signaling pathways may be involved in the pathology of AR. Furthermore, a circRNA-miRNA interaction network was constructed via miRNA response elements analysis. Some circRNAs were correlated with miRNAs that are involved in T cell polarization and activation, thereby highlighting their potential role in the pathogenesis of AR. Conclusions This study demonstrates a number of aberrantly expressed circRNAs related to AR, and offers a novel perspective into AR pathogenesis and future therapeutic strategies (AU)

Altered circular RNA expression profiles in an ovalbumin-induced murine model of allergic rhinitis.

BACKGROUND: Emerging evidence shows that circular RNAs (circRNAs) participate in the pathogenesis of multiple immune diseases. However, few studies have focused on the mechanisms of circRNAs involved in allergic rhinitis (AR). METHODS: This study performed an RNA sequence (RNA-seq) profiling to identify the expression of circRNAs in nasal mucosa from ovalbumin-induced AR murine models and normal controls. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) was then conducted to validate the differential expression of circRNAs. Bioinformatics analysis was applied to demonstrate the biological functions of the dysregulated circRNAs. RESULTS: A total of 86 distinct circRNA candidates were sequenced, of which 51 were upregulated and 35 were downregulated. The T cell receptor, B cell receptor, and calcium signaling pathways may be involved in the pathology of AR. Furthermore, a circRNA-miRNA interaction network was constructed via miRNA response elements analysis. Some circRNAs were correlated with miRNAs that are involved in T cell polarization and activation, thereby highlighting their potential role in the pathogenesis of AR. CONCLUSIONS: This study demonstrates a number of aberrantly expressed circRNAs related to AR, and offers a novel perspective into AR pathogenesis and future therapeutic strategies.

Insulinoma-associated protein 1 controls nasopharyngeal carcinoma to radiotherapy by modulating cyclin D1-dependent DNA repair machinery.

Insulinoma-associated protein 1 (INSM1), a zinc finger transcriptional factor, is proven to be deregulated in several types of cancers. However, comprehension of the molecular mechanism of INSM1-mediated tumor progression remains poor. Here, we show that the radioresistant nasopharyngeal carcinoma (NPC) patients have higher expressions of INSM1 that correlated with poor prognosis. Genetic manipulation of INSM1 expression sufficiently controls the response of NPC cells to irradiation (IR). Mechanistically, cells exposed to IR, increased intracellular INSM1 competitively disrupts the interaction of cyclin D1 and CDK4 resulting in cell survival by the cyclin D1-dependent DNA repair machinery. Moreover, knockdown of INSM1 sensitives NPC cells to IR in vivo and protects xenograft mice from mortality. Taken together, these results indicate that INSM1 modulates NPC to radiotherapy by controlling cyclin D1-dependent DNA repair machinery that could be manipulated as a novel molecular target for NPC therapy.

Composition and abundance of microbiota in the pharynx in patients with laryngeal carcinoma and vocal cord polyps.

The pharynx is an important site of microbiota colonization, but the bacterial populations at this site have been relatively unexplored by culture-independent approaches. The aim of this study was to characterize the microbiota structure of the pharynx. Pyrosequencing of 16S rRNA gene libraries was used to characterize the pharyngeal microbiota using swab samples from 68 subjects with laryngeal cancer and 28 subjects with vocal cord polyps. Overall, the major phylum was Firmicutes, with Streptococcus as the predominant genus in the pharyngeal communities. Nine core operational taxonomic units detected from Streptococcus, Fusobacterium, Prevotella, Granulicatella, and Veillonella accounted for 21.3% of the total sequences detected. However, there was no difference in bacterial communities in the pharynx from patients with laryngeal cancer and vocal cord polyps. The relative abundance of Firmicutes was inversely correlated with Fusobacteria, Proteobacteria, Actinobacteria, and Bacteroidetes. The correlation was evident at the genus level, and the relative abundance of Streptococcus was inversely associated with Fusobacterium, Leptotrichia, Neisseria, Actinomyces, and Prevotella. This study presented a profile for the overall structure of the microbiota in pharyngeal swab samples. Inverse correlations were found between Streptococcus and other bacterial communities, suggesting that potential antagonism may exist among pharyngeal microbiota.

Alterations of microbiota structure in the larynx relevant to laryngeal carcinoma.

The microbial communities that inhabit the laryngeal mucosa build stable microenvironments and have the potential to influence the health of the human throat. However, the associations between the microbiota structure and laryngeal carcinoma remain uncertain. Here, we explored this question by comparing the laryngeal microbiota structure in laryngeal cancer patients with that in control subjects with vocal cord polyps through high-throughput pyrosequencing. Overall, the genera Streptococcus, Fusobacterium, and Prevotella were prevalent bacterial populations in the laryngeal niche. Tumor tissue samples and normal tissues adjacent to the tumor sites (NATs) were collected from 31 laryngeal cancer patients, and the bacterial communities in laryngeal cancer patients were compared with control samples from 32 subjects. A comparison of the laryngeal communities in the tumor tissues and the NATs showed higher α-diversity in cancer patients than in control subjects, and the relative abundances of seven bacterial genera differed among the three groups of samples. Furthermore, the relative abundances of ten bacterial genera in laryngeal cancer patients differed substantially from those in control subjects. These findings indicate that the laryngeal microbiota profiles are altered in laryngeal cancer patients, suggesting that a disturbance of the microbiota structure might be relevant to laryngeal cancer.

The role of long non-coding RNA HOTAIR in the progression and development of laryngeal squamous cell carcinoma interacting with EZH2.

CONCLUSIONS: The aberrant expression of long non-coding RNA HOTAIR and transmethylase EZH2 played important roles in the progression and development of laryngeal squamous cell carcinoma (LSCC). OBJECTIVES: This research was aimed to explore the expression and correlation with clinicopathological characteristics of HOTAIR and EZH2 in LSCC, and to evaluate the function of the two in regulating the proliferation and cis-platinum resistance processes of LSCC. METHODS: Quantitative real time-PCR (qPCR) was conducted to measure the expression of HOTAIR and EZH2 in tissue samples. Clinicopathological features were collected and statistically analyzed combining with the expression of HOTAIR and EZH2. The variance of EZH2 with down-regulating HOTAIR was measured by qPCR. CCK-8 proliferation test was conducted to detect the proliferation feature in LSCC cells. After cultured with a series of cis-platinum concentrations for 24 h, cell viability was detected using CCK-8 assay, and the inhibition rates were calculated. RESULTS: HOTAIR and EZH2 were over-expressed in LSCC tissue. The higher expression was significantly related to T phase, pathological grades, and risk of lymphatic metastasis of LSCC. Suppressing HOTAIR expression stimulated EZH2 expressing, promoted the proliferation of AMC-HN8 cells, and increased the sensitivity to cis-platinum of the LSCC cells.

Effect of multidrug resistance 1/P-glycoprotein on the hypoxia-induced multidrug resistance of human laryngeal cancer cells.

In a previous study, it was demonstrated that hypoxia upregulated the multidrug resistance (MDR) of laryngeal cancer cells to chemotherapeutic drugs, with multidrug resistance 1 (MDR1)/P-glycoprotein (P-gp) expression also being upregulated. The present study aimed to investigate the role and mechanism of MDR1/P-gp on hypoxia-induced MDR in human laryngeal carcinoma cells. The sensitivity of laryngeal cancer cells to multiple drugs and cisplatin-induced apoptosis was determined by CCK-8 assay and Annexin-V/propidium iodide staining analysis, respectively. The accumulation of rhodamine 123 (Rh123) in the cells served as an estimate of drug accumulation and was evaluated by flow cytometry (FCM). MDR1/P-gp expression was inhibited using interference RNA, and the expression of the MDR1 gene was analyzed using reverse transcription-quantitative polymerase chain reaction and western blotting. As a result, the sensitivity to multiple chemotherapeutic agents and the apoptosis rate of the hypoxic laryngeal carcinoma cells increased following a decrease in MDR1/P-gp expression (P<0.05). Additionally, FCM analysis of fluorescence intensity indicated that the downregulated expression of MDR1/P-gp markedly increased intracellular Rh123 accumulation (P<0.05). Such results suggest that MDR1/P-gp serves an important role in regulating hypoxia-induced MDR in human laryngeal carcinoma cells through a decrease in intracellular drug accumulation.

MicroRNA-93 regulates cyclin G2 expression and plays an oncogenic role in laryngeal squamous cell carcinoma.

microRNA93 (miR-93) is expressed in the miR­106b-25 cluster, located in intron 13 of the MCM7 gene. Our previous study found that miR-93 was significantly upregulated in laryngeal squamous cell carcinoma (LSCC), and cyclin G2 (CCNG2) was a potential target of miR-93 in LSCC. However, the possible functions and molecular mechanisms of miR-93 in LSCC remain unknown. In the present study, we show that the level of CCNG2 protein expression was significantly lower in LSCC cancer tissue than normal tissues. The level of CCNG2 was correlated with clinical stages, lymph node metastasis and histological grade. We further show that the expression level of miR-93 was inversely correlated with CCNG2 expression in clinical specimens. Furthermore, gain-of-function assays revealed that miR-93 promoted cell proliferation, decreased apoptosis rates, induced cell cycle arrest and promoted cell migration and invasion, whereas silencing of miR-93 attenuated these carcinogenic processes. In addition, overexpression of miR-93 in Hep-2 cells could reduce the mRNA and protein levels of CCNG2, whereas silencing of miR-93 in Hep-2 cells significantly increased CCNG2 expression. A luciferase assay verified that miR-93 could bind to the 3' untranslated region of CCNG2. Importantly, ectopic expression of CCNG2 in miR-93 cells rescued the effect of miR-93 on LSCC proliferation. Knockdown of CCNG2 promoted cell proliferation resembling that of miR-93 overexpression. These findings demonstrated that miR-93 promotes tumor growth by directly suppressing CCNG2. Taken together, these results suggested that this newly identified miR-93-CCNG2 axis may be involved in LSCC proliferation and progression. Our findings provide novel potential targets for LSCC therapy and prognosis.

[Isolated sphenoid sinus disease: a clinical analysis of 159 cases].

OBJECTIVE: Aimed to observe and analyse the diagnostic and therapeutic methods of the isolated sphenoid sinus disease, achieve earlier diagnosis and timelier intervention for this easily neglected disease and decrease the occurrence of misdiagnoses. METHOD: A retrospective study was conducted in 159 patients with isolated sphenoid sinus disease. RESULT: Headache was the most common presenting symptom (79.87%,127/159). Among the 159 cases, 60 (37.74%) had mucocele, 44 (27.67%) isolated sphenoiditis, 31 (19.50%) fungal sinusitis, 5 (3.14%) polyp, 3 (1.89%) fibrous dysplasia, 2 (1.26%) inverted papilloma, 3 (1.89%) chordoma, 3 (1.89%) squamous carcinoma, 3 (1.89%) malignant lymphoma, 2 (1.26%) neuroendocrine carcinoma, 2 (1.26%) olfactory neuroblastoma, and 1 (0.63%) malignant fibrohistiocytoma. A follow-up of 10 months to 4 years post-surgery showed good prognosis in most of the patients who underwent surgical therapy. CONCLUSION: The sphenoid sinus disease is often vague and nonspecific in its clinical presentation. The most common clinical symptom is headache, followed by vision changes. Endoscopic sphenoidotomy is the primary therapy for isolated sphenoid sinus disease.

[Inflammatory myofibroblastic tumor in the maxillary sinus: a case report and review of literature].

OBJECTIVE: To investigate the clinical feature, diagnostic and therapeutic methods of inflammatory myofibroblastic tumor(IMT). METHOD: One case of IMT was reported and the relevant literatures were reviewed. RESULT: The computed tomography scan showed irregular soft tissue density shade and aggressive bone destruction with unclear boundary. Pathological findings showed variable numbers of inflammatory cells and myofibroblastic spindle cells. Tumor cells were immunoreactive for vimentin and smooth muscle actin, but negative for desmin et al. CONCLUSION: IMT of the maxillary sinus is very rare. The diagnosis of IMT base on histopathology and immunohistochemistry. The genesis and development of IMT result from chromosomal translocations that often cause an overexpression of anaplastic lymphoma kinase. IMT are clinical and pathological distinct entities and its biological behavior is still uncertain.