RESUMO
Epithelial-mesenchymal transition (EMT) is thought to be involved in the tissue remodeling and long-term inflammatory process of chronic sinusitis (CRS), but the driving mechanism is still unclear. Using high-resolution mass spectrometry, we performed a proteomic screen of CRS nasal mucosal tissue to identify differentially expressed proteins. Data are available via ProteomeXchange with identifier PXD030884. Specifically, we identified S100 calcium binding protein A4 (S100A4), an effective factor in inflammation-related diseases, and its downstream protein closely related to tissue fibrosis collagen type I alpha 1 chain (COL1A1), which suggested its involvement in nasal mucosal tissue remodeling. In addition, stimulation of human nasal epithelial cells (HNEpCs) with lipopolysaccharide (LPS) mimicked the inflammatory environment of CRS and showed that S100A4 is involved in regulating EMT and thus accelerating tissue remodeling in the nasal mucosa, both in terms of increased cell motility and overexpression of mesenchymal-type proteins. Additionally, we further investigated the regulation mechanism of S100A4 involved in EMT in CRS. Our research results show that in the inflammatory environment of CRS nasal mucosal epithelial cells, TCF-4 will target to bind to S100A4 and regulate its transcription. The transcription of S100A4 in turn affects the execution of the important signaling pathway in EMT, the Wnt/GSK-3ß/ß-catenin pathway, through the TCF-4/ß-catenin complex. In conclusion, this study confirmed that the expression of S100A4 was significantly increased during the progressive EMT process of CRS mucosal epithelial cells, and revealed that the transcriptional regulation of S100A4 plays an important role in the occurrence and development of EMT. This finding will help us to better understand the pathogenesis behind the remodeling in CRS patients, and identify target molecules for the treatment of CRS.
Assuntos
Transição Epitelial-Mesenquimal , Proteína A4 de Ligação a Cálcio da Família S100/metabolismo , Sinusite/metabolismo , Proteína 2 Semelhante ao Fator 7 de Transcrição/metabolismo , beta Catenina/metabolismo , Movimento Celular , Doença Crônica , Cadeia alfa 1 do Colágeno Tipo I/metabolismo , Células Epiteliais/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Mucosa Nasal/metabolismo , Proteômica/métodos , Via de Sinalização WntRESUMO
Abnormal remodeling of the nasal mucosal epithelium and persistent chronic inflammation are important pathological features of chronic sinusitis with nasal polyps (CRSwNPs). In order to explore the molecular regulation mechanism of CRSwNPs, we performed iTRAQ protein profile analysis on 18 clinical samples collected (9 patients with nasal polyps and 9 healthy patients) and found that S100A11, a Ca2+-binding protein, was significantly higher in CRSwNPs. Subsequently, we demonstrated that S100A11 was mainly located in nasal mucosal epithelial cells and is up-regulated in human nasal epithelial stem/progenitor cells (hNESPCs) from CRSwNPs patients and CRSwNPs epithelial cell model established with S. aureus. To determine the functional role of S100A11 and the signal pathways in epithelial cells, we constructed S100A11 overexpression vector, small interfering RNA, recombinant protein-S100A11 (rh-S100A11) and RAGE inhibitor (sRAGE). Results showed that upregulation of S100A11 inhibited epithelial cell viability and promoted apoptosis and inflammation, in addition, S100A11 can regulate the signal homeostasis of AMPK-STAT3 via RAGE mediation in epithelial cells. Our findings suggest that S100A11 is involved in CRSwNPs epithelial tissue remodeling and inflammatory response regulation and may be a useful target for CRSwNPs therapy.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Antígenos de Neoplasias/metabolismo , Células Epiteliais/metabolismo , Inflamação/patologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Pólipos Nasais/patologia , Proteínas S100/metabolismo , Fator de Transcrição STAT3/metabolismo , Sinusite/patologia , Adolescente , Adulto , Idoso , Apoptose , Linhagem Celular , Proliferação de Células , Criança , Doença Crônica , Células Epiteliais/microbiologia , Células Epiteliais/patologia , Humanos , Pessoa de Meia-Idade , Modelos Biológicos , Nariz/patologia , Transdução de Sinais , Staphylococcus aureus/fisiologia , Regulação para Cima , Adulto JovemRESUMO
Long noncoding RNAs (lncRNAs) have been identified as potential prognostic tools and therapeutic biomarkers for a variety of human cancers. However, the functional roles and underlying mechanisms of key lncRNAs affecting laryngeal squamous cell carcinomas (LSCCs) are largely unknown. Here, we adopted a novel subpathway strategy based on the lncRNA-mRNA profiles from the Cancer Genome Atlas (TCGA) database and identified the lncRNA deleted in lymphocytic leukemia 2 (DLEU2) as an oncogene in the pathogenesis of LSCCs. We found that DLEU2 was significantly upregulated and predicted poor clinical outcomes in LSCC patients. In addition, ectopic overexpression of DLEU2 promoted the proliferation and migration of LSCC cells both in vivo and in vitro. Mechanistically, DLEU2 served as a competing endogenous RNA to regulate PIK3CD expression by sponging miR-30c-5p and subsequently activated the Akt signaling pathway. As a target gene of DLEU2, PIK3CD was also upregulated and could predict a poor prognosis in LSCC patients. In conclusion, we found that the novel LSCC-related gene DLEU2 enhances the malignant properties of LSCCs via the miR-30c-5p/PIK3CD/Akt axis. DLEU2 and its targeted miR-30c-5p/PIK3CD/Akt axis may represent valuable prognostic biomarkers and therapeutic targets for LSCCs.
Assuntos
Carcinoma de Células Escamosas/genética , Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias Laríngeas/genética , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Longo não Codificante/metabolismo , Transdução de Sinais , Sequência de Bases , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Progressão da Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Humanos , Neoplasias Laríngeas/patologia , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Análise Multivariada , Invasividade Neoplásica , Metástase Neoplásica , Oncogenes , Prognóstico , RNA Longo não Codificante/genética , Regulação para Cima/genéticaRESUMO
The occurrence and development of osteoclasts can directly affect the severity of bone destruction in middle ear cholesteatoma. At the same time, cell communication between keratinocytes and fibroblasts can stimulate osteoclast differentiation. However, the molecular mechanism of osteoclast differentiation in cholesteatoma is still poorly understood. In this study, we try to isolate the exosomes of keratinocytes from patients with middle ear cholesteatoma, and explore the effects of keratinocyte-derived exosomes (Ker-Exo) on osteoclast differentiation by co-culturing Ker-Exo with fibroblasts and osteoclast precursor cells. As a result, we confirmed that Ker-Exo primed fibroblasts can up-regulate the expression of RANKL and promote osteoclast differentiation. We revealed that the effect of Ker-Exo depened on its miRNA-17 conponent. Analysis confirmed that miRNA-17 was down-regulated in Ker-Exo, and they can increase RANKL level in fibroblasts, thus promoting the differentiation of osteoclasts. In conclusions, we provide evidence that exosomes miRNA-17 secreted by keratinocytes in patients with middle ear cholesteatoma can up-regulate the expression of RANKL in fibroblasts and induce osteoclast differentiation.