PURPOSE: Bmi-1 is a Polycomb group member which participates in many physiological processes as well as in a wide spectrum of cancers. The aim of this study was to investigate Bmi-1 expression in non-small cell lung cancer (NSCLC) in respect to clinicopathological features and therapeutic outcomes. METHODS: Immunohistochemical staining for Bmi-1 was performed on tissue microarrays (TMAs) constructed from 179 formalin-fixed and paraffin-embedded NSCLC samples (106 squamous, 58 adeno-, and 15 large cell carcinomas). Data were subject to statistical analysis by SPSS. RESULTS: Overall evaluation of all tumor cases showed that 20 (11.43%) were negative, 37 (21.14%) showed weak, 65 (37.14%) moderate and 57 (32.57%) strong nuclear positivity for Bmi-1. Statistical analysis of our data revealed that the expression of Bmi-1 was significantly higher in stage III (P = 10(-6)) and stage IV (P = 10(-5)) tumors compared to stages I and II tumors. The administration of adjuvant chemotherapy significantly increased DFS at stage I and II patients who did not express Bmi-1 when compared to their Bmi-1 positive counterparts (P = 0.05). CONCLUSIONS: Our results suggest that Bmi-1 is significantly associated with progression of NSCLC and might serve as a prognostic marker of adverse disease outcome.

• MeSH
• čipová analýza proteinů MeSH
• čipová analýza tkání MeSH
• financování organizované MeSH
• imunohistochemie MeSH
• jaderné proteiny genetika   metabolismus   MeSH
• lidé MeSH
• nádory plic metabolismus   patologie   MeSH
• nemalobuněčný karcinom plic metabolismus   patologie   MeSH
• prognóza MeSH
• protoonkogenní proteiny genetika   metabolismus   MeSH
• represorové proteiny genetika   metabolismus   MeSH
• staging nádorů MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

microarrayAims: MRP-1, LRP and TOPO-II are all associated with protection of the cells from the adverse eff ects of variouschemotherapeutics. The aim of this study was to measure the expression of these proteins in Wilms' tumor (WT).Materials and Methods: TMA block was constructed from 14 samples of WT's and from xenografts derived fromthem. Sections of the TMA were used for immunostaining against MRP-1, LRP and TOPO-IIa.Results: All normal kidneys expressed MRP-1 but were either weakly or negatively stained for LRP and TOPO-IIa.In WT samples, MRP-1 was universally expressed, exclusively in the tubular component, while there was no expressionof LRP and TOPO-IIa showed heterogeneous distribution. The xenografts varied in their MRP-1 and TOPO-IIaexpression and exhibited weak/negative staining of LRP.Conclusions: This study shows that although all the proteins evaluated, had diff erent expression patterns in thetumor samples, the most prominent changes in expression were found for MRP-1. The exact clinical implications ofthese changes in expression and their relevance to the resistance of these tumors to chemotherapy requires furtherinvestigation. The fi nding of diff erent expression profi les for the multidrug resistance proteins in the original WT'sand their xenografts suggests that the results of animal cancer models may be diffi cult to interpret.

• MeSH
• čipová analýza tkání metody   MeSH
• DNA-topoisomerasy typu II genetika   imunologie   izolace a purifikace   MeSH
• exprese genu genetika   imunologie   MeSH
• financování organizované MeSH
• imunohistochemie metody   využití   MeSH
• medicína založená na důkazech MeSH
• modely u zvířat MeSH
• myši inbrední NOD MeSH
• myši SCID MeSH
• proteiny spojené s mnohočetnou rezistencí k lékům genetika   imunologie   izolace a purifikace   MeSH
• vault ribonucleoprotein particles genetika   imunologie   izolace a purifikace   MeSH
• Wilmsův nádor genetika   imunologie   MeSH
• xenogenní modely - testy protinádorové aktivity metody   využití   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

BACKGROUND: T cell density in colorectal cancer (CRC) has proven to be of high prognostic importance. Here, we evaluated the influence of a hyperfractionated preoperative short-term radiation protocol (25 Gy) on immune cell density in tumor samples of rectal cancer (RC) patients and on patient survival. In addition, we assessed spatial tumor heterogeneity by comparison of analogue T cell quantification on full tissue sections with digital T cell quantification on a virtually established tissue microarray (TMA). METHODS: A total of 75 RC patients (60 irradiated, 15 treatment-naïve) were defined for retrospective analysis. RC samples were processed for immunohistochemistry (CD3, CD8, PD-1, PD-L1). Analogue (score 0-3) as well as digital quantification (TMA: 2 cores vs. 6 cores, mean T cell count) of marker expression in 2 areas (central tumor, CT; invasive margin, IM) was performed. Survival was estimated on the basis of analogue as well as digital marker densities calculated from 2 cores (Immunoscore: CD3/CD8 ratio) and 6 cores per tumor area. RESULTS: Irradiated RC samples showed a significant decrease in CD3 and CD8 positive T cells, independent of quantification mode. T cell densities of 6 virtual cores approximated to T cell densities of full tissue sections, independent of individual core density or location. Survival analysis based on full tissue section quantification demonstrated that CD3 and CD8 positive T cells as well as PD-1 positive tumor infiltrating leucocytes (TILs) in the CT and the IM had a significant impact on disease-free survival (DFS) as well as overall survival (OS). In addition, CD3 and CD8 positive T cells as well as PD-1 positive TILs in the IM proved as independent prognostic factors for DFS and OS; in the CT, PD-1 positive TILs predicted DFS and CD3 and CD8 positive T cells as well as PD-1 positive TILs predicted OS. Survival analysis based on virtual TMA showed no impact on DFS or OS. CONCLUSION: Spatial tumor heterogeneity might result in inadequate quantification of immune marker expression; however, if using a TMA, 6 cores per tumor area and patient sample represent comparable amounts of T cell densities to those quantified on full tissue sections. Consistently, the tissue area used for immune marker quantification represents a crucial factor for the evaluation of prognostic and predictive biomarker potential.

• Publikační typ
• časopisecké články MeSH

Malignant gliomas are among the most severe types of cancer, and the most common primary brain tumors. Treatment options are limited and the prognosis is poor. WNT-5A, a member of the WNT family of lipoglycoproteins, plays a role in oncogenesis and tumor progression in various cancers, whereas the role of WNT-5A in glioma remains obscure. Based on the role of WNT-5A as an oncogene, its potential to regulate microglia cells and the glioma-promoting capacities of microglia cells, we hypothesize that WNT-5A has a role in regulation of immune functions in glioma. We investigated WNT-5A expression by in silico analysis of the cancer genome atlas (TCGA) transcript profiling of human glioblastoma samples and immunohistochemistry experiments of human glioma tissue microarrays (TMA). Our results reveal higher WNT-5A protein levels and mRNA expression in a subgroup of gliomas (WNT-5A(high)) compared to non-malignant control brain tissue. Furthermore, we show a significant correlation between WNT-5A in the tumor and presence of major histocompatibility complex Class II-positive microglia/monocytes. Our data pinpoint a positive correlation between WNT-5A and a proinflammatory signature in glioma. We identify increased presence of microglia/monocytes as an important aspect in the inflammatory transformation suggesting a novel role for WNT-5A in human glioma.

• MeSH
• čipová analýza tkání MeSH
• gliom metabolismus   patologie   MeSH
• lidé MeSH
• mikroglie metabolismus   patologie   MeSH
• monocyty metabolismus   patologie   MeSH
• proteiny Wnt biosyntéza   genetika   metabolismus   MeSH
• protoonkogenní proteiny biosyntéza   genetika   metabolismus   MeSH
• výpočetní biologie MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

AIMS: Invasive ductal and lobular carcinomas are the most common histological types of breast cancer. The loss of E-cadherin expression has been suggested to be the most reliable marker for invasive lobular carcinoma. The aim of our study was to identify the diagnostic usefulness of novel markers in the differentiation of these tumor types. METHODS: We examined tissue microarrays (TMA) which were constructed from surgical specimens of 119 breast cancer patients. TMA consisted of 80 ductal carcinomas, 29 lobular carcinomas and special type cancers. TMA sections were stained using standard immunohistochemical methods. Monoclonal mouse antibodies against E-cadherin, cytokeratin 5/6 and 17, and polyclonal mouse antibodies against EMP1, DDR1, PRKCI and DVL1 were used. RESULTS: E-cadherin was absent in 93.3% of lobular tumors compared with only 15 % of ductal tumors (p<0.0001). EMP1 and DVL1 were overexpressed in lobular tumors (93.1% and 96.5%, respectively), whereas PRKCI and DDR1 were positive in ductal cancers (90% and 96.2%, respectively). Reduced expression or absence of both cytokeratins 5/6 and 17 was found in both tumor tissues in comparison to normal terminal duct lobular units (p<0.0001). CONCLUSIONS: Apart from the well-established marker, E-cadherin, proteins examined on TMA slides by immunohistochemistry (EMP1, DVL1, DDR1, PRKCI) may represent novel tissue markers helpful in the differentiation of ductal and lobular breast cancers. Further studies with larger sets of patients are desirable, to verify the complete immunohistochemical profiles of various histological types of breast cancer and determine the prognostic and predictive significance of novel markers.

• MeSH
• čipová analýza tkání MeSH
• diferenciální diagnóza MeSH
• dospělí MeSH
• duktální karcinom prsu diagnóza   MeSH
• imunohistochemie MeSH
• lidé MeSH
• lobulární karcinom diagnóza   MeSH
• nádorové biomarkery analýza   MeSH
• nádory prsu diagnóza   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVES: LC3A protein is associated with autophagosomes, and LC3A immunohistochemistry (IHC) is used for the detection of autophagy activity. The aim of this study was to assess the prognostic value of LC3A expression in patients with resected non-small cell lung cancer (NSCLC). MATERIALS AND METHODS: We used tissue microarrays (TMAs) constructed from 116 resected stage IB-III NSCLC patients. Standard immunohistochemistry was performed on formalin-fixed paraffin-embedded tissue sections using antibody against LC3A autophagic potein. Stained slides were scanned by Olympus dotSlide Digital Virtual Microscopy System (Japan) and the LC3A staining was evaluated digitally. Groups were compared using the Mann Whitney U test, and correlations were assessed using Spearman's rank test. Survival was calculated using Kaplan-Meier analysis. Primary study endpoint was overall survival (OS), secondardy study endpoint disease-free survival (DFS). Cut-off optimization for LC3A prognostic value was performed using the "cut-off finder' 'software (Charite, Berlin, Germany). In addition, the Kaplan Meier plotter (KmPlot) was used to assess the relationship between LC3A mRNA expression and clinical outcome (OS and DFS) in patients with NSCLC. RESULTS: From 116 patients, 88 tissue samples were available for final examination. No significant association was found between LC3A staining and other clinicopathological variables, including tumor grade, stage and histological subtype. A higher number of LC3A stone-like structures (SLSs) (>20), was significanly associated with poor OS (HR = 2.27, p = 0.011) and DFS (HR = 2.27, p = 0.003). A significant association between high LC3A mRNA and both a worse OS and worse DFS was found by KMPlot analysis in patients with stage I-III NSCLC. CONSLUSION: This retrospective study suggests that SLSs as assessed by LC3A IHC as well as LC3A mRNA expression has a clinically relevant negative prognostic value in patients with resected NSCLC, and should be further investigated.

• MeSH
• analýza přežití MeSH
• autofagie MeSH
• čipová analýza tkání MeSH
• imunohistochemie MeSH
• lidé MeSH
• messenger RNA genetika   MeSH
• nádorové biomarkery metabolismus   MeSH
• nádory plic imunologie   mortalita   MeSH
• nemalobuněčný karcinom plic imunologie   mortalita   MeSH
• pneumektomie MeSH
• prognóza MeSH
• proteiny asociované s mikrotubuly genetika   metabolismus   MeSH
• staging nádorů MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Aims. No effective treatment for lung cancer exists currently. One reason for this, is the development of drug resistance,-assumed to be associated with cancer stem cell (CSCs) emergence within the tumour. This pilot study aimed-to identify CSCs in 121 non-small cell lung cancer (NSCLC) patient samples via detection of the expression of stem-cell markers – CD133 and nestin. Material and methods. Archived paraffin blocks of 121 patient samples were prepared as Tissue Microarrays-(TMA). Indirect immunohistochemical staining was used to determine the level of expression of CD133 and nestin.-Double immunofluorescence staining was used to investigate the co-expression of these two markers. To determine-the correlation between expression of nestin and CD133 with the length of asymptomatic period and overall patient-survival we used the Kaplan-Meyer analysis. Results. CD133 expression was detected in 22 (19%), nestin in the epithelium in 74 (66%) and vasculature in-78 (70%) of patients. Co-expression of these two markers was found in 21 (17%) patients in less than 1% of positive-cells without impact on disease free or overall survival. Conclusions. We identified CD133+/nestin+ cells as novel potential markers of lung cancer CSCs.

• MeSH
• CD antigeny MeSH
• cévy metabolismus   MeSH
• epitel metabolismus   MeSH
• financování organizované MeSH
• glykoproteiny metabolismus   MeSH
• lidé MeSH
• nádorové kmenové buňky metabolismus   MeSH
• nádory plic krevní zásobení   metabolismus   MeSH
• nemalobuněčný karcinom plic krevní zásobení   metabolismus   MeSH
• peptidy metabolismus   MeSH
• pilotní projekty MeSH
• proteiny intermediálních filament metabolismus   MeSH
• proteiny nervové tkáně metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH

We aimed to identify novel markers for aggressive prostate cancer in a STAT3-low proteomics-derived dataset of mitochondrial proteins by immunohistochemical analysis and correlation with transcriptomic data and biochemical recurrence in a STAT3 independent PCa cohort. Formalin-fixed paraffin-embedded tissue (FFPE) sample selection for proteomic analysis and tissue-microarray (TMA) generation was conducted from a cohort of PCa patients. Retrospective data analysis was performed with the same cohort. 153 proteins differentially expressed between STAT3-low and STAT3-high samples were identified. Out of these, 46 proteins were associated with mitochondrial processes including oxidative phosphorylation (OXPHOS), and 45 proteins were upregulated, including NDUFS1/ATP5O. In a STAT3 independent PCa cohort, high expression of NDUFS1/ATP5O was confirmed by immunocytochemistry (IHC) and was significantly associated with earlier biochemical recurrence (BCR). mRNA expression levels for these two genes were significantly higher in intra-epithelial neoplasia and in PCa compared to benign prostate glands. NDUFS1/ATP5O levels are increased both at the mRNA and protein level in aggressive PCa. Our results provide evidence that NDUFS1/ATP5O could be used to identify high-risk PCa patients.

• Publikační typ
• časopisecké články MeSH

BACKGROUNDS: SWI/SNF complexes represent a family of multi-subunit chromatin remodelers that are affected by alterations in >20% of human tumors. While mutations of SWI/SNF genes are relatively uncommon in prostate cancer (PCa), the literature suggests that deregulation of various subunits plays a role in prostate tumorigenesis. To assess SWI/SNF functions in a clinical context, we studied the mutually exclusive, paralogue accessory subunits SMARCD1, SMARCD2, and SMARCD3 that are included in every known complex and are sought to confer specificity. METHODS: Performing immunohistochemistry (IHC), the protein levels of the SMARCD family members were measured using a tissue microarray (TMA) comprising malignant samples and matching healthy tissue of non-metastatic PCa patients (n = 168). Moreover, IHC was performed in castration-resistant tumors (n = 9) and lymph node metastases (n = 22). To assess their potential role as molecular biomarkers, SMARCD1 and SMARCD3 protein levels were correlated with clinical parameters such as T stage, Gleason score, biochemical recurrence, and progression-free survival. RESULTS: SMARCD1 protein levels in non-metastatic primary tumors, lymph node metastases, and castration-resistant samples were significantly higher than in benign tissues. Likewise, SMARCD3 protein expression was elevated in tumor tissue and especially lymph node metastases compared to benign samples. While SMARCD1 levels in primary tumors did not exhibit significant associations with any of the tested clinical parameters, SMARCD3 exhibited an inverse correlation with pre-operative PSA levels. Moreover, low SMARCD3 expression was associated with progression to metastasis. CONCLUSIONS: In congruence with previous literature, our results implicate that both SMARCD1 and SMARCD3 may exhibit relevant functions in the context of prostate tumorigenesis. Moreover, our approach suggests a potential role of SMARCD3 as a novel prognostic marker in clinically non-metastatic PCa.

• MeSH
• chromozomální proteiny, nehistonové * genetika   metabolismus   MeSH
• imunohistochemie MeSH
• lidé středního věku MeSH
• lidé MeSH
• lokální recidiva nádoru patologie   metabolismus   genetika   MeSH
• lymfatické metastázy MeSH
• nádorové biomarkery * genetika   metabolismus   MeSH
• nádory prostaty rezistentní na kastraci patologie   genetika   metabolismus   MeSH
• nádory prostaty * patologie   metabolismus   genetika   MeSH
• prognóza MeSH
• senioři MeSH
• stupeň nádoru MeSH
• transkripční faktory genetika   metabolismus   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH

Expression of intrinsic markers of tumour hypoxia and angiogenesis are important predictors of radiotherapeutic, and possibly surgical, outcome in several cancers. Extent of tumour hypoxia in localised prostate cancer is comparable to that in other cancers, but few data exist on the association of extent of tumour hypoxia with treatment outcome. We aimed to study the predictive value of intrinsic markers of tumour hypoxia and angiogenesis in localised prostate cancer, both in patients treated with radiotherapy and in those treated surgically. METHODS: We applied a new, needle biopsy tissue microarray (TMA) technique to study diagnostic samples from men with localised, previously untreated prostate cancer treated in two randomised controlled trials of radiotherapy-dose escalation. Multivariate analysis by Cox proportional hazards was done to assess the association between clinical outcome, in terms of biochemical control, and immunohistochemical staining of hypoxia inducible factor-1 alpha (HIF-1 alpha), vascular endothelial growth factor (VEGF), and osteopontin expression. The analysis was repeated on an independent series of men with localised, previously untreated prostate cancer treated by radical prostatectomy. The main outcome was time to biochemical (ie, prostate-specific antigen [PSA]) failure. FINDINGS: Between Oct 12, 1995, and Feb 5, 2002, 308 patients were identified from two prospective, randomised trials at the Royal Marsden Hospital, London and Sutton, UK, for the radiotherapy cohort and diagnostic biopsies were available for 201 of these patients. Between June 6, 1995, and Nov 4, 2005, 329 patients were identified from the Aarhus University Hospital, Skejby, Denmark, for the prostatectomy cohort; of these, 40 patients were excluded because the tumour was too small to sample (19 patients), because the paraffin block was too thin (19 patients), or because the blocks were missing (two patients), leaving 289 patients for analysis. For patients treated with radiotherapy, increased staining for VEGF (p=0.008) and HIF-1 alpha (p=0.02) expression, but not increased osteopontin expression (p=0.978), were significant predictors of a shorter time to biochemical failure on multivariate analysis, independent of clinical tumour stage, Gleason score, serum PSA concentration, and dose of radiotherapy. For patients treated with surgery, increased staining for VEGF (p<0.0001) and HIF-1 alpha (p<0.0001) expression, and increased osteopontin expression (p=0.0005) were each significantly associated with a shorter time to biochemical failure on multivariate analysis, independent of pathological tumour stage, Gleason score, serum PSA concentration, and margin status. INTERPRETATION: To our knowledge, this is the largest study of intrinsic markers of hypoxia and angiogenesis in relation to the outcome of radical treatment of localised prostate cancer. Increased expression of VEGF, HIF-1 alpha, and, for patients treated with surgery, osteopontin, identifies patients at high risk of biochemical failure who would be suitable for enrolment into trials of treatment intensification.

• MeSH
• analýza přežití MeSH
• faktor 1 indukovatelný hypoxií genetika   metabolismus   MeSH
• hodnocení rizik MeSH
• hypoxie buňky MeSH
• imunohistochemie MeSH
• jehlová biopsie MeSH
• kohortové studie MeSH
• konformní radioterapie metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové biomarkery analýza   genetika   MeSH
• nádory prostaty MeSH
• osteopontin genetika   metabolismus   MeSH
• patologická angiogeneze genetika   metabolismus   MeSH
• pravděpodobnost MeSH
• prognóza MeSH
• proporcionální rizikové modely MeSH
• prostatektomie metody   MeSH
• prostatický specifický antigen krev   MeSH
• randomizované kontrolované studie jako téma MeSH
• referenční hodnoty MeSH
• retrospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• staging nádorů MeSH
• vaskulární endoteliální růstový faktor A genetika   metabolismus   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH