epidermal-growth-factor and Nasal-Polyps

The pathogenesis of chronic rhinosinusitis (CRS) remains unclear to date. The tissue remodeling in nasal polyps may be the result of inflammatory mediators and may involve epithelial-mesenchymal transition (EMT) and EMT-associated features such as cell motility in nasal epithelial cells (NECs). We determined whether NEC in nasal polyps of CRS already display features of EMT in vivo or respond with EMT to growth factor stimulation in vitro. Nasal polyp tissues expressed both epithelial and mesenchymal markers. Primary NEC from inferior turbinates and nasal polyps responded to the EMT-inducing agents transforming growth factor (TGF)-β1 and epidermal growth factor (EGF) with different expression patterns of EMT markers (E-cadherin, N-cadherin, Snail, Slug, Twist), however, only NEC from nasal polyps were susceptible to TGF-β1 and EGF-dependent cell migration. Our data suggest that a partial EMT is associated with the pathogenesis of nasal polyps in CRS patients. Furthermore, we show for the first time that epithelial cells from both nasal polyps and inferior turbinates were able to undergo an EMT-like process following exposure to TGF-β1 or EGF in vitro but that only NEC from nasal polyps responded with enhanced cell motility. Our data suggest that NEC from CRS patients have undergo partial EMT and that this process may be involved in the pathogenesis of CRS.

Topics: Adult; Aged; Aged, 80 and over; Cell Movement; Epidermal Growth Factor; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Gene Expression Profiling; Humans; Inflammation; Male; Middle Aged; Nasal Polyps; Oligonucleotide Array Sequence Analysis; Sinusitis; Transforming Growth Factor beta1; Turbinates; Young Adult

Epidermal growth factor receptors play an important role in airway epithelial cell growth and differentiation. The current study investigates the expression profiles of EGF, EGFR and ERBB4 in patients with nasal polyps (NP), and their response to glucocorticosteroid (GC) treatment. Fifty patients with NP (40 without GC treatment and 10 with oral GC) and 20 control subjects with septal deviation were recruited into the study. Protein levels of EGF, EGFR, and ERBB4 were evaluated by immune-staining. In healthy nasal epithelium, EGF and EGFR localized within p63+ basal cells, while ERBB4 localized within ciliated cells. GC-naïve NP epithelium showed weak expression of EGF in 90% of samples versus 5% of controls. EGFR was significantly increased in the epithelium with basal cell hyperplasia from GC-naïve NPs (78%, 31/40) compared to controls (23%, 4/17). EGFR was also found in some degranulating goblet cells. ERBB4 expression was significantly higher in hyperplastic epithelium from GC-naïve NPs (65%, 26/40) than in controls (6%, 1/17). GC treatment restored the EGF expression and normalized the EGFR and ERBB4 expression in NPs. Differential expression patterns of EGF, EGFR, and ERBB4 are essential in epithelial restitution and remodeling in nasal epithelium.

Topics: Adult; Case-Control Studies; Epidermal Growth Factor; ErbB Receptors; Female; Gene Expression Profiling; Humans; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Receptor, ErbB-4; Regeneration; Young Adult

A better understanding of the expression profile of a group of angiogenic markers in nasal polyps (NPs) would contribute considerably to the investigation of the formation of NPs. The aim of this study was to evaluate the combined mRNA expression of vascular endothelial growth factor A (VEGFA), fibroblast growth factor 2 (FGF2), transforming growth factor beta1 (TGFbeta1), epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF1) in NPs obtained from 21 patients undergoing nasal polypectomy. Nasal mucosae were obtained from the adjacent inferior turbinates (AIT) and middle turbinates (AMT) of the patients, as well as from 11 control subjects undergoing nasal corrective surgery. Analysis was performed using real-time RT-PCR. VEGFA, TGFbeta1 and IGF1 exhibited significant over-expression in the NPs compared to the control turbinates, EGF did not exhibit significant expression, and FGF2 presented constant over-expression in the NPs compared to both the adjacent and control turbinates. Since its mRNA levels were positively correlated with all the corresponding levels of the rest of the growth factors studied, TGFbeta1 seems to be a key cytokine in interactions between NP cells and the leading molecule of the epithelial differentiation and tissue remodelling present in the disease. Many correlations between the transcript levels of the other growth factors arose in the NP group as well, supporting a co-regulation of these genes in nasal polyposis. Our conclusions were that that VEGFA and TGFbeta1 participate significantly in the formation of NPs, whereas FGF2 and IGF1 are implicated in nasal polyposis to a lesser, but still significant, extent. EGF does not seem to be actively involved in the NP evolution process.

Topics: Adult; Aged; Epidermal Growth Factor; Female; Fibroblast Growth Factor 2; Gene Expression Profiling; Humans; Insulin-Like Growth Factor I; Intercellular Signaling Peptides and Proteins; Male; Middle Aged; Nasal Polyps; RNA, Messenger; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

To investigate the expression of epidermal growth factor receptor (EGFR) messenger RNA (mRNA) in human sinus mucosa and to compare the expression of EGFR and EGF among patients with chronic rhinosinusitis (CRS), patients with CRS and nasal polyposis (CRS/NP), and a healthy control group.. Maxillary sinus ostia mucosa was harvested from patients undergoing endoscopic sinus surgery for CRS or CRS/NP and from patients undergoing surgery for non-CRS pathologic conditions (control group). The samples were analyzed using semiquantitative reverse transcription-polymerase chain reaction to detect mRNA of EGFR. Hematoxylin-eosin staining and immunofluorescent staining were used to localize EGFR and EGF in the sinus mucosa.. Academic research.. Three groups (CRS, CRS/NP, and control), each with 10 subjects, were enrolled in the present study.. Area ratios of positive cells in the epithelia were compared among the CRS, CRS/NP, and control groups. In addition, eosinophils were counted in the subepithelial connective tissue in the 3 groups.. The level of EGFR mRNAs in the sinus mucosa of the CRS and CRS/NP groups was statistically significantly increased compared with that in the control group (P < .01), and no statistically significant difference was found between the sinus mucosa of the CRS group and that of the CRS/NP group (P < .01). On hematoxylin-eosin staining, hyperplasia and metaplasia of epithelial goblet cells were present in the sinus mucosa of the CRS and CRS/NP groups. Epidermal growth factor receptor was mainly expressed in goblet cells and basal cells and was weakly expressed in ciliated cells, while EGF expression was located in epithelial cells and in some inflammatory cells but not in goblet cells. In the control group, expression of EGFR and EGF was lower compared with that in the CRS and CRS/NP groups. No statistically significant area ratios of positive cells differences in staining of EGFR and EGF were found between the CRS group and the CRS/NP group (P > .05), whereas statistically significant differences were found between the control group and the 2 CRS groups (P < .01). The number of eosinophils was statistically significantly increased in the CRS/NP group compared with that in the CRS group (P < .01).. Up-regulation of the EGFR cascade may have an important role regarding mucus production in the sinus mucosa of patients with CRS and CRS/NP associated with hyperplasia and metaplasia of epithelial goblet cells.

Topics: Case-Control Studies; Chronic Disease; Eosinophils; Epidermal Growth Factor; ErbB Receptors; Humans; Leukocyte Count; Nasal Mucosa; Nasal Polyps; Rhinitis; RNA, Messenger; Sinusitis

To elucidate a mechanism of proliferation and differentiation of nasal gland cells, we established a serum-free 3-dimensional culture system for human nasal gland (HNG) cells and examined the effects of epidermal growth factor, keratinocyte growth factor, and retinoic acid on proliferation and differentiation of cultured HNG cells.. Nasal polyps were obtained from patients undergoing endoscopic endonasal sinus surgery. The HNG cells were cultured under a monolayer culture and transferred to a collagen-embedded culture using RPMI 1640 medium containing transferrin, insulin, hydrocortisone, retinoic acid, epidermal growth factor, and keratinocyte growth factor. Cell growth was measured by bromodeoxyuridine incorporation assays. To measure cell differentiation, the percentage of cells containing secretory granules, which were stained with Alcian blue in cytoplasm, was determined.. In the serum-free 3-dimensional culture, the HNG cells showed ductal structures containing secretory products in a lumen. The addition of epidermal growth factor promoted the proliferation of HNG cells in its optimal concentrations, and keratinocyte growth factor also enhanced the proliferation of HNG cells. Conversely, the differentiation of HNG cells was not dependent on epidermal growth factor and keratinocyte growth factor. Retinoic acid suppressed the proliferation, but promoted the differentiation of HNG cells.. Our culture system could be useful for studying the effects of various growth factors and cytokines on HNG proliferation and differentiation to better understand the mechanisms of growth and morphogenesis of nasal glands.

Topics: Cells, Cultured; Epidermal Growth Factor; Fibroblast Growth Factor 7; Fibroblast Growth Factors; Humans; In Vitro Techniques; Nasal Mucosa; Nasal Polyps; Tretinoin

Human surface respiratory epithelial (HSRE) cells from nasal polyps have been cultured within collagen lattices in a serum-free defined medium. Cell growth observed over a period of 12 days showed a population doubling time of 36 h. Under these culture conditions, we observed a contraction of the lattices. Phase-contrast light microscopy and transmission electron microscopy demonstrated that the HSRE cells formed tubular ductlike structures. Lumens formed by HSRE cells were surrounded by cuboidal-shaped polarized cells with numerous ciliated cells, secretory cells, and undifferentiated cells. Epidermal growth factor (EGF) was observed to stimulate the tubule formation and the contraction of the lattices. Videomicroscopic observations and analysis of the ciliary beating frequency (CBF) demonstrated that the cilia were homogeneously distributed on the whole apical surface of the ciliated cells and that their movement was well coordinated, with a CBF similar to that observed in outgrowth cells from cultured human nasal and tracheal epithelia. Immunofluorescent staining of basement membrane components synthesized and secreted by cells revealed the presence of type III collagen around the tubules. Type IV collagen and laminin were present in the cytoplasm and at the periphery of the cells. The biotin-streptavidin-gold immunocytochemical technique with monoclonal anti-mucin antibody showed intracellular localization of mucins in secretory granules of the secretory cells. With the use of substrate gel electrophoresis polyacrylamide gels impregnated with gelatin, collagenase activity was detected in the conditioned medium of the cultured HSRE cells. These results suggest that both three-dimensional collagen gel and soluble factors such as EGF regulate tubule formation by HSRE cells. Moreover, the capacity of the epithelial cells to contract the gel suggests they may be involved in the wound healing process.

Topics: Basement Membrane; Cell Division; Cell Separation; Cells, Cultured; Cilia; Collagen; Collagenases; Culture Media, Serum-Free; Epidermal Growth Factor; Gels; Humans; Kinetics; Microscopy, Electron; Mucus; Nasal Mucosa; Nasal Polyps

The Cystic Fibrosis antigen (CFA) is a 14 kDa. Ca2(+)-binding protein known to be expressed in cells of myeloid origin during normal cell differentiation. CFA serum levels are elevated in Cystic Fibrosis (CF) patients and heterozygotes. We examined the expression of CFA in different cultured epithelial cells from controls and patients with CF. The steady state level of CFA was in general higher in epithelial cells from CF patients compared to control cells and was found to increase during cell aging. The latter difference could be attributed to an increased rate of CFA synthesis rather than to an impairment of CFA degradation or secretion, as shown by pulse chase experiments.

Topics: Blood Proteins; Calgranulin A; Cells, Cultured; Cystic Fibrosis; Cytoskeletal Proteins; Epidermal Growth Factor; Epithelium; Granulocytes; Humans; Immunoblotting; Kinetics; Molecular Weight; Nasal Polyps; Reference Values