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  • 1
    Keywords: CANCER ; Germany ; human ; PROSTATE ; THERAPY ; DIAGNOSIS ; GENE ; GENES ; PROTEIN ; PROTEINS ; validation ; MARKER ; BIOMARKERS ; TARGET ; IDENTIFICATION ; PROGRESSION ; prostate cancer ; PROSTATE-CANCER ; FUSION ; MARKERS ; PREDICTION ; TARGETS ; HUMAN GENES ; molecular biology ; molecular ; THERAPIES ; TISSUE MICROARRAYS ; LEVEL ; biomarker ; analysis ; HIGH-THROUGHPUT ; technique ; CANCERS ; CHALLENGES ; TARGETED THERAPY ; CLINICAL-PRACTICE ; gene protein
    Abstract: Significant cellular alterations required for the development and progression of cancers are detectable at the molecular level and represent potential targets for gene-specific therapies. Modern chip techniques allow the parallel analysis of virtually all known human genes and proteins in a single experiment. Using modern high-throughput techniques, numerous potential new biomarkers for the diagnosis and prediction of prostate cancer have been identified. However, so far none of these markers has improved clinical practice. One of the most important challenges in the coming years is the extensive clinical validation of molecular data using clinically relevant end points. For this venture the pivotal prerequisite is the availability of large, comprehensively annotated and standardized high-quality bioresources
    Type of Publication: Journal article published
    PubMed ID: 18712514
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  • 2
    Keywords: CANCER ; EXPRESSION ; Germany ; human ; PROSTATE ; THERAPY ; DIAGNOSIS ; GENE ; GENES ; PROTEIN ; PROTEINS ; validation ; BIOMARKERS ; IDENTIFICATION ; PROGRESSION ; prostate cancer ; PROSTATE-CANCER ; FUSION ; TARGETS ; SINGLE ; molecular biology ; THERAPIES ; TISSUE MICROARRAYS ; development ; HIGH-THROUGHPUT ; USA ; CANCERS ; TARGETED THERAPY ; ANNEXIN A3
    Abstract: Significant cellular alterations required for the development and progression of cancers are detectable at the molecular level and represent potential targets for gene-specific therapies. Modern chip techniques allow the parallel analysis of virtually all known human genes and proteins in a single experiment. Using modern high-throughput techniques, numerous potential new biomarkers for the diagnosis and prediction of prostate cancer have been identified. However, so far none of these markers has improved clinical practice. One of the most important challenges in the coming years is the extensive clinical validation of molecular data using clinically relevant end points. For this venture the pivotal prerequisite is the availability of large, comprehensively annotated and standardized high-quality bioresources
    Type of Publication: Journal article published
    PubMed ID: 19139898
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  • 3
    Keywords: EXPRESSION ; GROWTH ; carcinoma ; PATHWAY ; PROTEINS ; transcription ; GROWTH-FACTOR-BETA ; prostate carcinoma ; PROTEASOME ; MULTIPLE-MYELOMA ; TRANSFORMING GROWTH-FACTOR-BETA-1 ; BREAST-CANCER CELLS ; BORTEZOMIB ; DEPENDENT DEGRADATION ; E3 ubiquitin ligase ; SMAD2 ; SMURF2 ; tumour marker
    Abstract: The purpose of this work was to investigate the role of the ubiquitin-proteasome network (UPN) in prostate cancer (PCA) and to elicit potential markers for this disease. The UPN represents a key factor in the maintenance of cellular homoeostasis as a result of its fundamental function in the regulation of intracellular protein degradation. Members of this network have a role in the biology of haematological and solid tumours. Tumour cells and normal epithelial cells from 22 prostatectomy specimens were isolated by laser microdissection. Prostate biopsy samples from healthy individuals served for technical calibration and as controls. Transcript levels of eight selected genes with E3 ubiquitin ligase activity (labelling target proteins for proteasome degradation) and two genes belonging to the proteasome-multienzyme complex itself were analysed by quantitative real-time RT-PCR. The proteasome genes PSMC4 and PSMB5 and the E3 ubiquitin ligase NEDD4L were significantly and coherently upregulated in PCA cells compared with the corresponding adjacent normal prostate tissue. Transcription of the E3 ubiquitin ligase SMURF2 was significantly higher in organ-confined tumours (pT2) compared with non-organ-confined cancers (pT3). The results indicate a role for PSMC4 and PSMB5 and the E3 ubiquitin ligase NEDD4L in prostate tumourigenesis, whereas SMURF2 downregulation could be associated with clinical progression. NEDD4L and SMURF2 both target transforming growth factor (TGF)-beta for degradation. This reflects the pleiotropic role of the TGF-beta signalling pathway acting as a tumour suppressor in normal and pre-cancerous cells, but having oncogenic properties in progressing cancer. Further studies have to elucidate whether these alterations could represent clinically relevant PCA-diagnostic and progression markers.
    Type of Publication: Journal article published
    PubMed ID: 21102547
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  • 4
    Keywords: CANCER ; CELL ; Germany ; QUANTIFICATION ; GENOME ; PROTEIN ; PROTEINS ; validation ; LINES ; BIOMARKERS ; CELL-LINES ; PHOSPHORYLATION ; antibodies ; antibody ; TARGET ; PERFORMANCE ; AMPLIFICATION ; microarrays ; ARRAYS ; CELL-LINE ; LINE ; Jun ; STRATEGIES ; sensitivity ; cell lines ; IMMUNOASSAYS ; signaling ; HUMAN CANCER ; proteome ; methods ; high throughput ; HIGH-THROUGHPUT ; PHASE ; SPECIMENS ; STRATEGY ; RANGE ; CATALYZED REPORTER DEPOSITION ; STROMAL-TUMORS GISTS
    Abstract: Background: Reverse phase protein arrays (RPPA) emerged as a useful experimental platform to analyze biological samples in a high-throughput format. Different signal detection methods have been described to generate a quantitative readout on RPPA including the use of fluorescently labeled antibodies. Increasing the sensitivity of RPPA approaches is important since many signaling proteins or posttranslational modifications are present at a low level. Results: A new antibody-mediated signal amplification ( AMSA) strategy relying on sequential incubation steps with fluorescently-labeled secondary antibodies reactive against each other is introduced here. The signal quantification is performed in the near-infrared range. The RPPA-based analysis of 14 endogenous proteins in seven different cell lines demonstrated a strong correlation (r = 0.89) between AMSA and standard NIR detection. Probing serial dilutions of human cancer cell lines with different primary antibodies demonstrated that the new amplification approach improved the limit of detection especially for low abundant target proteins. Conclusions: Antibody-mediated signal amplification is a convenient and cost-effective approach for the robust and specific quantification of low abundant proteins on RPPAs. Contrasting other amplification approaches it allows target protein detection over a large linear range
    Type of Publication: Journal article published
    PubMed ID: 20569466
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  • 5
    Keywords: THERAPY ; MORTALITY ; PROTEINS ; BIOMARKERS ; TARGET ; PROGRESSION ; PROTEOMIC ANALYSIS ; SUPPRESSOR ; GENE-EXPRESSION DATA ; FUSIONS
    Abstract: The present study aimed to investigate the proteome profiling of surgically treated prostate cancers. Hereto, 2D-DIGE and mass spectrometry were performed for protein identification, and data validation for peroxiredoxin 3 and 4 (PRDX3 and PRDX4) was accomplished by reverse phase protein arrays (RPPA). The Formal Concept Analysis (FCA) method was applied to assess whether the TMPRSS2-ERG gene fusion could influence the degree of overexpression of PRDX3 and PRDX4 in prostate cancer. Lastly, we performed an in vitro functional characterization of both PRDX3 and PRDX4 using the classical human prostate cancer cell lines DU145 and LNCaP. Reverse phase protein arrays verified that the overexpression of both PRDX3 and PRDX4 in tumor samples is negatively correlated with the presence of the TMPRSS2-ERG gene fusion. Functional characterization of PRDX3 and PRDX4 activity in PCa cell lines suggests a role of these members of the peroxiredoxin family in the pathophysiology of this tumor entity.
    Type of Publication: Journal article published
    PubMed ID: 22424448
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