Mir-142-3p Regulates Inflammatory Response by Contributing to Increased TNF-α in Chronic Rhinosinusitis With Nasal Polyposis.

OBJECTIVE: Previous studies suggested that microRNAs played an important role in the progression of inflammation and remodeling of chronic rhinosinusitis with nasal polyposis. However, the abnormal expression of microRNAs and regulation cytokine expression in nasal polyposis are not clear. METHOD: The miR-142-3p and tumor necrosis factor α (TNF-α) expression levels in chronic rhinosinusitis with nasal polyposis were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The miR-142-3p and TNF-α levels in human nasal epithelial cells (HNEpC) after stimulation by lipopolysaccharide (LPS) were detected by qRT-PCR. Moreover, HNEpCs were transfected by miR-142-3p mimics or inhibitor or cotransfected with si-TNF-α to evaluate the regulation of miR-142-3p on TNF-α which affects the production of inflammatory factors. RESULTS: The miR-142-3p and TNF-α were significantly higher in nasal mucosa of chronic rhinosinusitis with polyps patients compared to normal human. MiR-142-3p and TNF-α expression levels were increased after LPS stimulation in a dose- and time-dependent manner. Knockdown of miR-142-3p in HNEpCs downregulated TNF-α expression at both messenger RNA and protein levels. CONCLUSIONS: It is indicated that miR-142-3p may participate in the regulation of the body's inflammatory response through the LPS-TLR-TNF-α signaling pathway in chronic rhinosinusitis with nasal polyposis.

Knockdown of lncRNA ANRIL suppresses the production of inflammatory cytokines and mucin 5AC in nasal epithelial cells via the miR-15a-5p/JAK2 axis.

The incidence of allergic rhinitis (AR) is increasing worldwide. Human nasal epithelial cells (HNECs) are the key cells in the occurrence of AR. Antisense non-coding RNA in the INK4 locus (ANRIL) was discovered to be involved in the progression of AR. However, the mechanism by which ANRIL mediates the progression of AR remains to be determined. The present study aimed to further explore the mechanism by which ANRIL regulates AR. Thereby, HNECs were treated with IL-13 to mimic AR in vitro. The mRNA expression levels of ANRIL, microRNA (miR)-15a-5p, JAK2, mucin 5AC (MUC5AC), granulocyte-macrophage colony-stimulating factor (GM-CSF) and eotaxin-1, and protein expression levels of JAK2, STAT3 and phosphorylated-STAT3 in HNECs were analyzed using reverse transcription-quantitative PCR and western blotting, respectively. ELISAs were used to detect the secretory levels of inflammatory cytokines and mucin in cell supernatants. In addition, a dual luciferase reporter assay was used to confirm the downstream target of ANRIL and the target gene of miR-15a-5p. The results revealed that the secretory levels of eotaxin-1, GM-CSF and MUC5AC were significantly upregulated by IL-13 in the supernatant of HNECs. The expression levels of ANRIL and JAK2 were also upregulated in IL-13-induced HNECs, while the expression levels of miR-15a-5p were downregulated. In addition, ANRIL was identified to bind to miR-15a-5p. The IL-13-induced upregulation of eotaxin-1, GM-CSF and MUC5AC mRNA expression and secretory levels was significantly inhibited by the genetic knockdown of ANRIL, while the miR-15a-5p inhibitor effectively reversed this effect. JAK2 was also discovered to be directly targeted by miR-15a-5p. The overexpression of JAK2 significantly suppressed the therapeutic effect of miR-15a-5p mimics on IL-13-induced inflammation in vitro. In conclusion, the findings of the present study suggested that the genetic knockdown of ANRIL may suppress the production of inflammatory cytokines and mucin in IL-13-treated HNECs via regulation of the miR-15a-5p/JAK2 axis. Thus, ANRIL may serve as a novel target for AR treatment.

microRNA-18a from M2 Macrophages Inhibits TGFBR3 to Promote Nasopharyngeal Carcinoma Progression and Tumor Growth via TGF-ß Signaling Pathway.

OBJECTIVES: Nasopharyngeal carcinoma (NPC) is a type of nasopharyngeal disease with high metastasis and invasion properties. Tumor-associated alternative activated (M2) macrophages are evidenced to connect with NPC. Based on this, this study purposes to explore the mechanism and participation of microRNA-18a (miR-18a) from M2 macrophages in NPC. METHODS: Peripheral blood mononuclear cells were differentiated to macrophages and macrophages were polarized to M2 type by interleukin-4. SUNE-1 and CNE2 cells were transfected with restored or depleted miR-18a or transforming growth factor-beta III receptor (TGFBR3) to explore their roles in NPC progression with the involvement of the TGF-ß signaling pathway. Next, SUNE-1 and CNE2 cells were co-cultured with M2 macrophages that had been treated with restored or depleted miR-18a or TGFBR3 to comprehend their combined roles in NPC with the involvement of the TGF-ß signaling pathway. RESULTS: MiR-18a was highly expressed and TGFBR3 was lowly expressed in NPC cells. MiR-18a restoration, TGFBR3 knockdown or co-culture with miR-18a mimics, or si-TGFBR3-transfected M2 macrophages promoted SUNE-1 cell progression, tumor growth in mice, decreased p-Smad1/t-Smad1, and elevated p-Smad3/t-Smad3. miR-18a downregulation, TGFBR3 overexpression, or co-culture with miR-18a inhibitors or OE-TGFBR3-transfected M2 macrophages depressed CNE2 cell progression, tumor growth in mice, increased p-Smad1/t-Smad1, and decreased p-Smad3/t-Smad3. CONCLUSION: Our study elucidates that miR-18a from M2 macrophages results in promoted NPC cell progression and tumor growth in nude mice via TGFBR3 repression, along with the Smad1 inactivation and Smad3 activation.

LINC00669 insulates the JAK/STAT suppressor SOCS1 to promote nasopharyngeal cancer cell proliferation and invasion.

Nasopharyngeal carcinoma (NPC) is an epithelial cancer emerging from the lining of nasopharyngeal mucosa, with extremely frequent occurrence in east and southeast Asia. For the purpose of exploring roles of the dysregulated long non-coding RNA (lncRNA) in NPC, we identified a novel lncRNA LINC00669 with an apparent negative correlation to the overall survival from human NPC mRNA expression profiling databases. We further performed RNA pulldown coupled with mass spectrum to find out its target protein, and applied a series of in vitro and in vivo loss-and-gain-of function assays to investigate its oncogenic roles in NPC tumor development and progression. Our results demonstrated that LINC00669 competitively binds to the key JAK/STAT signaling pathway suppressor SOCS1, and insulates it from imposing ubiquitination modification on the pathway component of STAT1, which leads to its abnormal stabilization and activation. The activated STAT1 is then transferred into the nucleus and initiates the transcription of genes related to proliferation and invasion. In summary, our study reveals that the cytoplasmic resident lncRNA LINC00669 confers malignant properties on NPC cancer cells by facilitating a persistent activation of the JAK/STAT signaling pathway. Findings in the current study shed lights on prospects for treating NPC using strategies targeting the novel regulator of the JAK/STAT signaling.

Endoscopic endonasal transsphenoidal approach for sellar tumors beyond the sellar turcica.

CONCLUSIONS: The endoscopic endonasal transsphenoidal approach can be a choice for sellar tumors beyond the sellar turcica, but it is necessary to make the choice carefully because of the severe surgical risks. OBJECTIVES: To summarize our experience of removal of sellar tumors beyond the sellar turcica via the endoscopic endonasal transsphenoidal approach and to evaluate the surgical efficacy and complications. METHODS: Between January 2007 and January 2012, 30 patients with sellar tumors beyond the sellar turcica underwent surgery using the endoscopic endonasal transsphenoidal approach. RESULTS: Postoperative pathological examination demonstrated that pituitary adenoma occurred in 22 patients, craniopharyngioma in 5, and meningioma in 3. Total removal was achieved in 21 patients (70.0%) and subtotal removal was achieved in 8 patients (26.7%). After the surgery, cerebrospinal fluid leakage occurred in 3 patients, temporary diabetes insipidus occurred in 25 patients and persistent diabetes insipidus in 4 patients, intracranial infection occurred in 1 patient, frontal subdural effusion occurred in 1 patient, sinusitis occurred in 2 patients, epistaxis occurred in 3 patients, and 1 patient with a huge pituitary adenoma died of hypothalamic failure related to the operation.

[Inflammatory cytokine expression in recurrent nasal polyps by antibody chips].

OBJECTIVE: To investigate the expression of inflammatory cytokines in patients with recurrent nasal polyps by antibody chips. METHODS: Proteins from the patients'nasal membrane in a nasal polyps group, a recurrent nasal polyps group, and a control group were labeled with biotin. The biotin-labeled proteins reacted with antibody chips on which the antibodies of 40 major inflammatory cytokines were prepared. The target proteins were conjugated with streptomycin antibody labeled with horseradish peroxidase (HRP),and signals were imaged by laser scanner. RESULTS: Compared with the control group, the levels of inflammatory cytokines of nasal polyp group were notably increased, including pro-and anti-inflammatory cytokines, chemokines and certain cytokine receptors; while in recurrent nasal polyps, expression of chemokines were increased and most anti-inflammatory cytokines were decreased. CONCLUSION: Antibody chips demonstrate a significant change in cytokine profiles in patients with recurrent nasal polypsis, as compared with those with nasal polyps. The abnormally higher expression of chemotatic factors in the nasal mucosa may play an important role in the recurrence of human nasal polyps.

[Expression and significance of human beta-defensin in recurrent nasal polyps].

OBJECTIVE: To observe the expression of human beta-defensin-1 (hBD-1) and human beta-defensin-2 (hBD-2) in recurrent nasal polyps, and to investigate the role of beta-defensin in the recurrent nasal polyps. METHODS: Tissues of nasal polyps was obtained from 10 patients with nasal polyps undergoing endoscopic sinus surgery, recurrent nasal polyps from 10 patients 6 months after surgery, nasal mucosa from 10 recovered patients with nasal polyps postoperatively and,10 control subjects. hBD-1 mRNA and hBD-2 mRNA levels of tissue specimens in all groups were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: There was no significant difference in hBD-1 mRNA level between the 4 groups (P>0.05). Expression of hBD-2 mRNA was detected in patients with nasal polyps and recurrent nasal polyps, but rare in the recovered patients and the control subjects. CONCLUSION: hBD-1 is a constitutive expression and hBD-2 is an induced expression. beta-Defensin may play an important role in forming the nasal polyps.

[Establishment of 2-dimensional gel electrophoresis map and analysis of proteomics from human nasal polyps].

OBJECTIVE: To establish 2-dimensional polyacrylamide gel electrophoresis (2-DE) map from human nasal polyps and normal nasal mucosa, and to identify differential expression proteins of 2-DE map. METHODS: Samples of nasal polyps and nasal mucosa (each sample group containing 7 cases) were obtained. The total proteins were extracted and separated by immobilized pH gradient (IPG)-based 2-DE. The silver-stained 2-DE was scanned with digital Imagescanner and analyzed with ImageMaster 2-DE Elite 4.01 software. To obtain peptide mass fingerprint (PMF) of differential protein spots, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used. The PMF was searched in Swiss-Prot and TreMBL database by Pept-Ident software, to identify differential expression proteins. RESULTS: The well-resolved, reproducible 2-DE maps of nasal polyps and nasal mucosa were established. For the polyps tissues, the average proteins spot of three 2-DE maps was 825+/-78; and 682+/-96 spot was matched with the average matching rate of 82.7%. The average deviations of matched spot position were (1.13+/-0.16) mm in IEF direction and (1.45+/-0.21) mm in SDS-PAGE direction, respectively. For the nasal mucosa tissues, the average proteins spot of three 2-DE maps was 936+/-62; and 821+/-78 spots were matched with the average matching rate of 87.7%. After comparing the 2-DE maps of nasal polyps and nasal mucosa tissues, the protein spots were 1,458 and 1,617 respectively; and 1,026 protein spots were matched. Forty differential expression protein spots were incised from silver staining gel randomly and digested in the gel by TPCK-Trypsin. Thirty-four PMFs were obtained by MALDI-TOF-MS and 24 differential proteins were identified. CONCLUSION: The well-resolved, reproducible 2-DE maps of human nasal polyps and nasal mucosa have been successfully established. Certain differential proteins related to the pathogenesis of human nasal polyps are identified.

[Differential proteins analysis for human nasal polyposis and normal nasal mucosa].

OBJECTIVE: To establish two-dimensional polyacrylamide gel electrophoresis (2-DE) map with high resolution and reproducibility from human nasal polyposis and normal nasal mucosa, and to identify differential expression proteins of 2-DE map. METHOD: Samples of human nasal polyposis and normal nasal mucosa (each sample group containing seven cases) were obtained. The total proteins were extracted and separated by immobilized pH gradient (IPG)-based 2-DE. The silver-stained 2-DE were scanned with digital Image Scanner and analyzed with ImageMaster 2-DE Elite 4.01 software. Peptide mass fingerprint (PMF) of differential protein spots was obtained with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The peptide mass fingerprints were searched in Swiss-Prot and TreMBL database by PeptIdent software, and then differential expression proteins were identified. RESULT: (1) 2-DE for a randomly selected 1 sample from each of the 2 groups was repeated 3 times respectively to analyze the reproductive of the method. The image analysis showed: For the polyposis tissues, the average proteins spots of three 2-DE maps were 891 +/- 67 and 767 +/- 83, spots were matched with the average matching rate of 86.1%. The average deviations of matched spot position were (1.13 +/- 0.16) mm in IEF direction and 1.45 +/- 0.21) mm in SDS-PAGE direction, respectively. For the nasal mucosa tissues, the average proteins spots of three 2-DE maps were 936 +/- 62 and 821 +/- 78, spots were matched with the average matching rate of 87.7%. Comparing the average electrophoresis profiles of the two tissues, each sample contained 7 cases. The proteins spots of nasal polyposis and nasal mucosa tissues were 1532 and 1617. A total of 1 065 proteins spots were matched between the two tissues average electrophoresis profiles. (2)Twenty dif-ferential expression protein spots were incised from silver staining gel randomly and digested in gel by TPCK Trypsin. Sixteen PMF were obtained by MALDI-TOF-MS, and 11 differential expression proteins were identified. CONCLUSION: In this study, the well-resolved reproducible 2-DE map of human nasal polyposis and nasal mucosa were established. Certain differential proteins were identified.