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  • 1
    Keywords: ANGIOGENESIS ; CELLS ; ENDOTHELIAL-CELLS ; EXPRESSION ; IN-VITRO ; proliferation ; CELL ; Germany ; human ; MODEL ; VITRO ; DISEASE ; SITE ; SITES ; DISTINCT ; GENE ; GENE-EXPRESSION ; GENES ; microarray ; PROTEINS ; transcription ; SURGERY ; NF-KAPPA-B ; ACTIVATION ; RESPONSES ; DNA ; TRANSCRIPTION FACTOR ; INDUCTION ; KERATINOCYTES ; BINDING ; cell cycle ; CELL-CYCLE ; CYCLE ; treatment ; cytokines ; IDENTIFICATION ; PATTERNS ; gene expression ; microarrays ; PROMOTER ; UP-REGULATION ; NUMBER ; PROMOTERS ; STRESS ; DNA-REPAIR ; REPAIR ; EXTRACELLULAR-MATRIX ; BETA ; ADHESION ; HUMAN-PAPILLOMAVIRUS TYPE-16 ; NF-kappa B ; DNA repair ; inflammation ; CDNA MICROARRAY ; CYTOKINE ; MATRIX ; RE ; extracellular matrix ; endothelial cell ; endothelial cells ; INTERLEUKIN-1 ; GENE-TRANSCRIPTION ; INTEGRINS ; analysis ; PROFILES ; BINDING-SITE ; INTERCELLULAR-ADHESION ; ONSET ; CANDIDATE ; UNIT ; BINDING-SITES ; ENDOTHELIAL-CELL ; PROINFLAMMATORY CYTOKINES ; cDNA arrays ; CERVICAL KERATINOCYTES ; mRNA gene transcription ; nuclear factor kappa-B ; PERIODONTAL-DISEASES
    Abstract: Proinflammatory cytokines such as interleukin-1 beta are known to be synthesized in oral gingivitis and periodontitis and lead to the activation of the transcription factor nuclear factor-kappa B (NF-kappa B). Although numerous effects of interleukin-1 beta on mesenchymal cells are known, e.g. up-regulation of intercellular adhesion moelcule-1 in endothelial cells, little is known of the effects of interleukin-1 beta on oral keratinocytes. The purpose of the present study was to seek interleukin-1 beta-mediated alterations in mRNA gene transcription and a putative activation of NF-kappa B in oral gingival keratinocytes. As an in vitro model for gingivitis and periodontitis, immortalized human gingival keratinocytes (IHGK) were stimulated with the proinflammatory cytokine interleukin-1 beta. An epithelia-specific cDNA microarray was used to analyze mRNA expression profiles from IHGK cells treated with 200 units interleukin-1 beta/ml for 3, 6, 9, 12, and 24 h. Indirect immunofluorescence was carried out to detect NF-kappa B in IHGK following interleukin-1 beta treatment. Detailed analysis revealed distinct patterns of time-dependent changes, including genes induced or repressed early (3-6 h) or late (12-24 h) after interleukin-1 beta treatment. Differentially expressed genes were involved in (i) cell stress, (ii) DNA repair, (iii) cell cycle and proliferation, (iv) anti-pathogen response, (v) extracellular matrix turnover, and (vi) angiogenesis. A large number of genes were responsive to NF-kappa B and induction was concomitant with nuclear translocation of the p65 RelA subunit of NF-kappa B. Interestingly, many of these genes contain multiple NF-kappa B binding sites in their promoters. Analysis of altered gene expression allows identification of gene networks associated with inflammatory responses. In addition to a number of well-known genes involved in gingivitis and periodontitis, we identified novel candidates that might be associated with the onset and maintenance of an inflammatory disease
    Type of Publication: Journal article published
    PubMed ID: 16953820
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  • 2
    Keywords: CANCER ; EXPRESSION ; tumor ; carcinoma ; COMBINATION ; Germany ; DISEASE ; DISEASES ; CDNA ; GENE ; GENE-EXPRESSION ; GENES ; microarray ; PROTEIN ; PROTEINS ; DIFFERENTIATION ; TISSUE ; FAMILY ; CONTRAST ; KERATINOCYTES ; SKIN ; DYNAMICS ; IDENTIFICATION ; PROGRESSION ; immunohistochemistry ; PATTERNS ; gene expression ; microarrays ; MEMBRANE ; mass spectrometry ; MASS-SPECTROMETRY ; cytoskeleton ; HEAD ; NECK ; squamous cell carcinoma ; CDNA MICROARRAYS ; CDNA MICROARRAY ; MASSES ; keratinocyte ; HNSCC ; ULTRAVIOLET ; S100 PROTEINS ; transcript ; CELL-CARCINOMA ; EXPRESSION PATTERNS ; ENVELOPE ; protein biomarkers ; S100 and annexin protein family ; SELDI-TOF MS
    Abstract: The calcium-binding proteins of the S100 and the annexin protein families have been implicated in a variety of important physiological functions including membrane remodeling, calcium-related Intracellular signaling, cytoskeleton dynamics. tissue homeostasis, and formation of the cornified envelope in differentiating keratinocytes. Deregulated expression of members of these families has been reported in different types of neoplasia and other diseases, but the results were not consistent. Here we have applied a combination of cDNA microarrays, quantitative reverse transcriptase-PCR (qRT-PCR) and surface enhanced laser desorption ionisation-time of flight mass spectrometry (SELDI-TOF MS) to Study differential expression of these genes in head and neck squamous cell carcinoma (HNSCC). The calgranulins A and B and annexins 1 and 2 were found to be down-regulated in HNSCC, compared with normal mucosa, at both the mRNA and protein level. Upon validation of the differential gene expression by tissue microarray immunohistochemistry, we detected novel expression patterns of calgranulins A and B both in normal mucosa as well as in HNSCC. In contrast to squamous cancer of skin and other cancers in which the calgranulins were found to be up-regulated, most HNSCC showed reduced and widely deranged staining patterns including heterogeneous nuclear, cytoplasmic and membranous staining, and even enhanced staining in the tumor stroma. These observations suggest that the normal function of the calgranulins A and B in mucosa might be different from that in skin. (c) 2005 Elsevier GmbH. All rights reserved
    Type of Publication: Journal article published
    PubMed ID: 15819419
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  • 3
    Keywords: CANCER ; EXPRESSION ; INHIBITOR ; tumor ; carcinoma ; CELL ; Germany ; CLASSIFICATION ; DIAGNOSIS ; RISK ; PROTEIN ; SAMPLES ; PATIENT ; BIOLOGY ; ASSOCIATION ; FIELD ; ALPHA ; STAGE ; IDENTIFICATION ; IN-SITU ; immunohistochemistry ; genetics ; ABERRATIONS ; MASS-SPECTROMETRY ; ONCOGENE ; HUMAN-PAPILLOMAVIRUS ; SQUAMOUS-CELL CARCINOMA ; HEAD ; NECK ; RISK ASSESSMENT ; heredity ; BIOPSY ; protein expression ; PROTEOMICS ; PROTEOMIC ANALYSIS ; NECK-CANCER ; CELL CARCINOMA ; ONCOLOGY ; TUMORIGENESIS ; ARRAY ; HNSCC ; LYMPH-NODE METASTASIS ; development ; analysis ; PROFILES ; EVENTS ; protein biomarkers ; HISTOLOGY ; FRAGMENT ; SELDI-TOF-MS ; BIOPSIES ; CLINICAL-IMPLICATIONS ; aberration ; comparison ; acyl-CoA-binding protein ; CANCERIZATION ; field cancerization ; GENETICALLY ALTERED FIELDS ; HUMAN NEUTROPHIL PEPTIDE-1 ; TUMOR-DISTANT EPITHELIA
    Abstract: Development of head and necksquamous cell carcinoma (HNSCC) is a multistep process and in many cases involves a phenomenon coined 'field cancerization'. In order to identify changes in protein expression occurring at different stages of tumorigenesis and field cancerization, we analysed 113 HNSCCs and 73 healthy, 99 tumor-distant and 18 tumor-adjacent squamous mucosae by SELDI-TOF-MS on IMAC30 ProteinChip Arrays. Forty-eight protein peaks were differentially expressed between healthy mucosa and HNSCC. Calgizarrin (S100A11), the Cystein proteinase inhibitor Cystatin A, Acyl-CoA-binding protein, Stratifin (14-3-3 sigma), Histone H4, alpha- and beta-Hemoglobin, a C-terminal fragment of beta-hemoglobin and the alpha-defensins 1-3 were identified by mass spectrometry. The alpha-defensins showed various alterations in expression as validated by immunohistochemistry (IHC). Supervised prediction analysis revealed excellent classification of healthy mucosa (94.5% correctly classified) and tumor samples (92.9% correctly classified). Application of this classifier to the tumor-adjacent and tumor-distant mucosa samples disclosed dramatic changes: only 59.6% of the tumor-distant biopsies were classified as normal, 27.3% were predicted as aberrant or HNSCC. Strikingly, 72% of the tumor-adjacent mucosae were predicted as aberrant. These data provide evidence for the existence of genetically altered fields with inconspicuous histology. Comparison of the protein profiles in the tumor-distant-samples with clinical outcome of 32 patients revealed a significant association between aberrant profiles with tumor relapse events (P = 0.018; Fisher's exact test, two-tailed). We conclude that proteomic pro. ling in conjunction with protein identification greatly outperforms histopathological diagnosis and may have significant predictive power for clinical outcome and personalized risk assessment
    Type of Publication: Journal article published
    PubMed ID: 16819514
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  • 4
    Abstract: We reasoned that micro-dissection of tumour cells for protein expression studies should be omitted since tumour-stroma interactions are an important part of the biology of solid tumours. To study such interactions in head and neck squamous cell carcinoma (HNSCC) development, we generated reference protein spectra for normal squamous epithelium and connective tissue by SELDI-TOF-MS. Calgranulins A and B, Annexin1 and Histone H4 were found to be strongly enriched in the epithelium. The alpha-defensins 1-3 and the haemoglobin subunits were identified in the connective tissue. Tumour-distant epithelia, representing early pre-malignant lesions, showed up-regulated expression of the stromal alpha-defensins, whereas the epithelial Annexin 1 was down-regulated. Thus, tumour microenvironment interactions occur very early in the carcinogenic process. These data demonstrate that omitting micro-dissection is actually beneficial for studying changes in protein expression during development and progression of solid tumours.
    Type of Publication: Journal article published
    PubMed ID: 21080850
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    Keywords: EXPRESSION ; SURVIVAL ; carcinoma ; CELL ; Germany ; CLASSIFICATION ; DIAGNOSIS ; FOLLOW-UP ; RISK ; SITE ; SITES ; GENES ; PROTEIN ; TISSUE ; TUMORS ; PATIENT ; IMPACT ; IDENTIFICATION ; REGION ; RECURRENCE ; REGIONS ; SQUAMOUS-CELL CARCINOMA ; HEAD ; NECK ; pathology ; relapse ; PROTEOMICS ; PROTEOMIC ANALYSIS ; NECK-CANCER ; CELL CARCINOMA ; ONCOLOGY ; HNSCC ; PROFILES ; prospective ; SELDI-TOF-MS ; SQUAMOUS-CELL ; PROFILE ; field cancerization ; tumours ; HEAD-AND-NECK ; Follow up ; proteomic ; biomarker protein profiles ; CHROMOSOME-17 ; ORAL EPITHELIAL DYSPLASIA ; pharynx and oesophagus carcinoma
    Abstract: 'Field cancerization' in head and neck squamous cell carcinoma (HNSCC) is poorly understood and it may extend from the pharynx into the oesophagus. Both local recurrences and second primary carcinomas/second field tumours may originate from field cancerization. Our prospective pilot study aimed at the identification of patients suffering from field cancerization on the basis of mucosal protein profiles. Five mucosal biopsies from the oropharynx, hypopharynx and from three regions of the oesophagus were taken from 24 patients. Protein profiles were generated from the mucosal biopsies. After classifier learning, using the profiles of the patients without tumour diagnosis (n = 9), we were able to discriminate between the different mucosal sites and between healthy mucosa and HNSCC using tumour and healthy tissue samples. Mucosal biopsies of tumour patients (n = 15) revealed changes in the protein profiles similar to those in the tumours. During 42 months median follow-up, six tumour patients experienced local recurrences and second field tumours, of which three occurred in the oesophagus. In all six cases, tumour relapse was correctly predicted by altered mucosal protein profiles (p = 0.007, Fisher's exact test, two-tailed). Consequently, molecular field cancerization had a strong impact on progression-free survival (p = 0.007, log-rank test). Protein profiles of small diagnostic biopsies hold great promise to improve personalized risk assessment in HNSCC. Larger studies are needed to further substantiate these findings. Copyright (C) 2010 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd
    Type of Publication: Journal article published
    PubMed ID: 20593486
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