The present study aimed to evaluate the pro-apoptotic effects of bleomycin A5 on nasal polyp‑derived fibroblasts (NPDFs) and the underlying molecular mechanisms. Nasal polyp tissue was acquired from 10 patients during surgery and NPDFs were isolated from surgical tissues. Fibroblasts were identified using immunohistochemistry. Bleomycin A5 was used to treat NPDFs along a concentration gradient. Cell viability was evaluated using a Cell Counting Kit‑8 assay. A flow cytometric Annexin V‑fluorescein isothiocyanate/propidium iodide assay was used to determine the percentage of apoptotic NPDFs. The mRNA expression levels of apoptotic genes were determined by reverse transcription‑quantitative polymerase chain reaction and levels of proteins associated with apoptosis were determined by western blotting. The results indicated that bleomycin A5 was able to induce apoptosis in NPDFs in a dose‑dependent manner. NPDFs treated with bleomycin A5 were identified to contain significantly high amounts of the active forms of caspase‑3 and showed considerable cleavage of poly(ADP‑ribose) polymerase. The mRNA and protein expression levels of the pro‑apoptotic molecule Bcl‑2‑associated X protein were significantly higher in treated NPDFs than in untreated NPDFs. In contrast, the mRNA and protein expression of the anti‑apoptotic molecule B‑cell lymphoma 2 (Bcl‑2) was significantly lower in treated NPDFs. These results indicated that bleomycin A5 could induce apoptosis in primary NPDFs through activation of the Bcl‑2 family and caspase cascades in a time-, and concentration-dependent manner.

Background: Much a ention on the pathophysiology of nasal polyp (NP) has focused on eosinophils. Interleukin (IL)-4 and eotaxin-3 (C-C motif chemokine ligand 26, or CCL26) levels have been reported to be increased in eosinophilic nasal polyps. The aim of this study was to characterize CCL26 pos ranscriptional regulation by the RNAbinding protein HuR in primary human nasal polyp-derived epithelial cells (hNPDECs) challenged with IL-4. Methods:A prospective, observational study was conducted. Nasal polyp tissues were obtained from eosinophilic (n = 12) and non-eosinophilic (n = 10) NP patients, and inferior turbinate (IT) tissues were taken from control subjects (n = 9) and cultured into hNPDECs. Expression of HuR and CCL26 were measured by immunohistochemistry, Western blot analysis, enzyme-linked immunoassay, and real-time polymerase chain reaction (PCR). The nucleocytoplasmic shu ling of HuR in hNPDECs was detected by immunofluorescence. Pos ranscriptional regulation of CCL26 by HuR was tested by ribonucleoprotein immunoprecipitation assay (RIP) and dual-luciferase reporter assay. CCL26 mRNA stabilization was measured by quatititative PCR a er treatment with actinomycin D. Student's t test and one-way analysis of variance were used.Results: Immunohistochemical data show that both HuR and CCL26 were highly expressed in NP tissues, especially eosinophilic NP tissues (p < 0.05). IL-4 stimulation increased CCL26 mRNA stability, and overexpression and knockdown of HuR affected CCL26 expression. Immunofluorescence data indicate that IL-4 altered the subcellular distribution of HuR. The RIP and dual-luciferase reporter assay results supply strong evidence for HuR binding to CCL26. Conclusion:Our results provide strong support for the hypothesis that IL-4-induced expression of CCL26 in hN-PDECs relies partly on CCL26 mRNA stabilization mediated by the interaction of HuR with CCL26 3'UTR. C 2018 ARS-AAOA, LLC.

BackgroundLiquid biopsy facilitates the enrichment and isolation of circulating tumor cells (CTCs) in various human cancers, including nasopharyngeal carcinoma (NPC). Characterizing CTCs allows observation of the evolutionary process of single tumor cells undergoing blood-borne dissemination, such as epithelial-mesenchymal transition. However, the prognostic value of phenotypic classification of CTCs in predicting the clinical outcomes of NPC remains poorly understood.Patients and methodsA total of 92 patients who met the inclusion criteria were enrolled in the present study. The CanPatrol™ CTC technology platform was employed to isolate CTCs, and an RNA in situ hybridization-based system was used for phenotypic classification. Kaplan–Meier survival curves were used for univariate survival analysis, and the log-rank test was performed for between-group comparisons of the survival curves.ResultsCTCs were detected in 88.0% (81/92) of the enrolled patients with NPC. The total CTC number did not vary between the T and N stages or between Epstein–Barr virus DNA-positive and -negative cases. The numbers of total CTCs and epithelial/mesenchymal (E/M) hybrid CTCs decreased significantly at 3 months post concurrent chemoradiotherapy (P=0.008 and P=0.023, respectively), whereas the numbers of epithelial or mesenchymal CTCs did not decrease. E/M hybrid-predominant cases had lower disease-free survival (P=0.043) and distant metastasis-free survival (P=0.046) rates than non-E/M hybrid-predominant cases.ConclusionCTC classification enables a better understanding of the cellular phenotypic alterations responsible for locoregional invasion and distant metastasis in NPC. E/M hybrid-predominant CTC distribution predicts unfavorable clinical outcomes in patients with progressive NPC.

After several local injections of BLE, the nasal polyp tissues decreased and then disappeared. During follow-up of 3 years the recurrence rate of this group was significantly lower than another one treated with operation plus medicine treatment. Apoptosis in BLE-treated tissue was prominently detected in the infiltrating inflammatory cells. The expression of PARP and casp-8 were increased in BLE-treated nasal polyp tissue compared with PBS-treated tissue.