miRNA expression in triple-negative breast cancers (TNBC) has mainly been studied from a methodological viewpoint. However, it has not been considered that miRNA expression profile may be associated with a specific morphological entity inside every tumor. The verification of this hypothesis on a set of 25 TNBCs was the subject of our previous work, where we confirmed specific expression of the studied miRNAs in 82 samples of different morphologies including inflammatory infiltrate, spindle cell, clear cell, and metastases after RNA extraction and purification as well as microchip and biostatistical analysis. In the current work, we demonstrate a low suitability of in situ hybridization method for miRNA detection compared to RT-qPCR, and in detail discuss the biological role of 8 miRNAs with the most significant changes of expression.
- MeSH
- lidé MeSH
- mikro RNA * genetika metabolismus MeSH
- triple-negativní karcinom prsu * genetika patologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- mikro RNA * MeSH
AIMS: The aim of this study was to compare the expression profile of selected DNA methyltransferases and global DNA methylation status in patients with different phases of multiple myeloma (MM) . For the analysis, different cellular populations including unsorted myeloma cells and a set of plasma cells purified by relevant antibodies were used. Consequently, laboratory data were compared to patients' clinical data. PATIENTS AND METHODS: For the analysis, unsorted bone marrow cell population of 44 MM patients (30 newly diagnosed, 9 relapsed and 5 patients in remission) and a set of 8 patients' samples of sorted plasma cells were used. We used commercially available RNA isolated from BM of 3 healthy individuals as control samples. Expression analysis of three DNA methyltransferases - DNMT1, DNMT3A, and DNMT3B was performed by quantitative RT-PCR and the patient global DNA methylation profiles were detected by colorimetric assay. RESULTS: Unchanged DNMT1 expression was detected in the selected cohort of patients. Normalized DNMT3A gene expression was globally higher in comparison with controls in unsorted and sorted cell populations. Low (0.08-1.81%) global DNA methylation status in unsorted samples of multiple myeloma patients did not correlate either with expression profiles of monitored DNA methyltransferases or with the stages of MM based on Durie-Salmon and International Staging System. CONCLUSION: This is the first comparative study between DNA methyltransferases expression profiles and global DNA methylation status in different phases of multiple myeloma patients. No significant correlation between the level of global methylation and the clinical stage of the unsorted cell population of patients with multiple myeloma was registered. Overexpression of the DNMT3A gene occurred in both sorted and unsorted cell populations of patients with multiple myeloma. This fact highlights the DNMT3A as a potential marker of multiple myeloma tumor progression. Moreover, we demonstrated comparable results in the expression of DNA methyltransferases in both sorted and unsorted cell populations. This is a promising result from the methodical point of view because when compared to samples of unsorted multiple myeloma cells, samples of sorted cells bring reduction of the number of possible analyses performed.
- Klíčová slova
- DNA methylation, DNA methyltransferases, multiple myeloma,
- MeSH
- DNA methyltransferasa 3A MeSH
- DNA MeSH
- lidé MeSH
- metylace DNA * MeSH
- mnohočetný myelom * genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA methyltransferasa 3A MeSH
- DNA MeSH
BACKGROUND: Meningioma is the most common primary central nervous system neoplasm, accounting for about a third of all brain tumors. Because their growth rates and prognosis cannot be accurately estimated, biomarkers that enable prediction of their biological behavior would be clinically beneficial. OBJECTIVE: To identify coding and noncoding RNAs crucial in meningioma prognostication and pathogenesis. METHODS: Total RNA was purified from formalin-fixed and paraffin-embedded tumor samples of 64 patients with meningioma with distinct clinical characteristics (16 recurrent, 30 nonrecurrent with follow-up of >5 years, and 18 with follow-up of <5 years without recurrence). Transcriptomic sequencing was performed using the HiSeq 2500 platform (Illumina), and biological and functional differences between meningiomas of different types were evaluated by analyzing differentially expression of messenger RNA (mRNA) and long noncoding RNA (IncRNA). The prognostic value of 11 differentially expressed RNAs was then validated in an independent cohort of 90 patients using reverse transcription quantitative (real-time) polymerase chain reaction. RESULTS: In total, 69 mRNAs and 108 lncRNAs exhibited significant differential expression between recurrent and nonrecurrent meningiomas. Differential expression was also observed with respect to sex (12 mRNAs and 59 lncRNAs), World Health Organization grade (58 mRNAs and 98 lncRNAs), and tumor histogenesis (79 mRNAs and 76 lncRNAs). Lnc-GOLGA6A-1, ISLR2, and AMH showed high prognostic power for predicting meningioma recurrence, while lnc-GOLGA6A-1 was the most significant factor for recurrence risk estimation (1/hazard ratio = 1.31; P = .002). CONCLUSION: Transcriptomic sequencing revealed specific gene expression signatures of various clinical subtypes of meningioma. Expression of the lnc-GOLGA61-1 transcript was found to be the most reliable predictor of meningioma recurrence.
- MeSH
- lidé MeSH
- lokální recidiva nádoru * diagnóza genetika MeSH
- meningeální nádory * diagnóza genetika MeSH
- meningeom * diagnóza genetika MeSH
- prognóza MeSH
- RNA dlouhá nekódující * genetika metabolismus MeSH
- stanovení celkové genové exprese MeSH
- transkriptom MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- RNA dlouhá nekódující * MeSH
Mucosal surfaces are colonized by highly diverse commensal microbiota. Coating with secretory IgA (SIgA) promotes the survival of commensal bacteria while it inhibits the invasion by pathogens. Bacterial coating could be mediated by antigen-specific SIgA recognition, polyreactivity, and/or by the SIgA-associated glycans. In contrast to many in vitro studies, only a few reported the effect of SIgA glycans in vivo. Here, we used a germ-free antibody-free newborn piglets model to compare the protective effect of SIgA, SIgA with enzymatically removed N-glycans, Fab, and Fc containing the secretory component (Fc-SC) during oral necrotoxigenic E. coli O55 challenge. SIgA, Fab, and Fc-SC were protective, whereas removal of N-glycans from SIgA reduced SIgA-mediated protection as demonstrated by piglets' intestinal histology, clinical status, and survival. In vitro analyses indicated that deglycosylation of SIgA did not reduce agglutination of E. coli O55. These findings highlight the role of SIgA-associated N-glycans in protection. Further structural studies of SIgA-associated glycans would lead to the identification of those involved in the species-specific inhibition of attachment to corresponding epithelial cells.
- MeSH
- aglutinace MeSH
- Escherichia coli fyziologie MeSH
- glykosylace MeSH
- gnotobiologické modely MeSH
- imunoglobulin A sekreční metabolismus MeSH
- imunoglobuliny - Fab fragmenty metabolismus MeSH
- infekce vyvolané Escherichia coli imunologie MeSH
- jednořetězcové protilátky metabolismus MeSH
- novorozená zvířata MeSH
- odolnost vůči nemocem MeSH
- polysacharidy metabolismus MeSH
- prasata MeSH
- těhotenství MeSH
- zvířata MeSH
- Check Tag
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- imunoglobulin A sekreční MeSH
- imunoglobuliny - Fab fragmenty MeSH
- jednořetězcové protilátky MeSH
- polysacharidy MeSH
BACKGROUND/AIM: Multiple myeloma is a highly heterogeneous disease of clonal plasma cells. Histone deacetylase (HDAC) inhibitors are promising anticancer drugs but their precise mechanisms of actions are not well understood. MATERIALS AND METHODS: Cell-cycle regulation and pro-apoptotic effects of two histone deacetylase inhibitors, suberohydroxamic acid (SAHA) and suberoylanilide hydroxamic acid (SBHA), were analyzed in multiple myeloma cell lines RPMI8226 and U266 with differing TP53 status using gene-expression analysis. RESULTS: Enhanced expression of cyclin-dependent kinase inhibitor 1A (CDKN1A/p21WAF/CIP1) detected in the TP53-deleted U266 cell line after SAHA treatment indicates the P53-independent mode of transcriptional activation of CDKN1A gene. In contrast, CDKN1A gene expression was significantly increased by both SBHA and SAHA treatment of TP53-mutated RPMI8226 cells. CONCLUSION: SAHA appears to be a potentially effective pro-apoptotic and anticancer drug with universal application in the treatment of heterogeneous populations of multiple myeloma cells.
- Klíčová slova
- Multiple myeloma, apoptosis, gene expression, myeloma cell lines,
- MeSH
- apoptóza účinky léků MeSH
- inhibitor p21 cyklin-dependentní kinasy antagonisté a inhibitory genetika MeSH
- inhibitory histondeacetylas farmakologie MeSH
- kontrolní body buněčného cyklu účinky léků MeSH
- kyseliny hydroxamové farmakologie MeSH
- lidé MeSH
- mnohočetný myelom farmakoterapie genetika metabolismus patologie MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 genetika metabolismus MeSH
- protinádorové látky farmakologie MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- CDKN1A protein, human MeSH Prohlížeč
- inhibitor p21 cyklin-dependentní kinasy MeSH
- inhibitory histondeacetylas MeSH
- kyseliny hydroxamové MeSH
- nádorový supresorový protein p53 MeSH
- protinádorové látky MeSH
- TP53 protein, human MeSH Prohlížeč
OBJECTIVES: DNA repair proteins have emerged as potential predictors for immunotherapy response alongside PD-L1 expression, tumor-infiltrating lymphocytes (TILs) and tumor mutational burden. We analyzed expression of PD-L1, TILs count and expression of the homologous recombination (HR) protein RAD51, as potential prognostic factors in patients with resected non-small-cell lung carcinoma (NSCLC). MATERIALS AND METHODS: Discovery set included 96 NSCLC patients from the University Hospital Olomouc (Czech Republic) and a replication set included 1109 NSCLC patients from University Hospital Zurich (Switzerland). Tissue microarrays (TMAs) were stained using the automated staining platform Ventana Benchmark Ultra with antibodies against RAD51,CD3, CD8, CD68 and PD-L1. RESULTS: Loss of nuclear RAD51 protein was associated with high TILs (r=-0.25, p = 0.01) and PD-L1 status (10.6 vs. 2.4 %, p = 0.012) in patients receiving neoadjuvant chemo-/radiotherapy (CT/RT). In silico analysis from the TCGA data set showed a negative relationship between RAD51 mRNA expression and CD45 (r = ‒0.422, p < 0.0001), CD68 (r = ‒0.326, p < 0.001), CD3 (r = ‒0.266, p < 0.001) and CD8 (r = ‒0.102, p < 0.001). RAD51 low/PD-L1 high patients were clustered as separate entity in the replication set and in TCGA dataset. High TILs status was significantly associated with improved OS in the replication set (unadjusted HR = 0.57, 95 % CI 0.42-0.76, p < 0.001). Similar results have been seen for CD3, CD8 and CD68. CONCLUSIONS: In conclusion, RAD51 nuclear loss is weakly associated with increased TILs and high PD-L1 at the time of surgery in curatively resected NSCLC and after prior exposure to neoadjuvant chemo- or radiotherapy. Both high TILs and RAD51 nuclear loss were confirmed as independent prognostic factors in curatively resected NSCLC.
- Klíčová slova
- Neoadjuvant, PD-L1, Prognostic, RAD51, Tumor infiltrating lymphocytes,
- MeSH
- antigeny CD274 genetika MeSH
- lidé MeSH
- nádory plic * genetika terapie MeSH
- nemalobuněčný karcinom plic * genetika terapie MeSH
- oprava DNA MeSH
- prognóza MeSH
- rekombinasa Rad51 genetika MeSH
- tumor infiltrující lymfocyty MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Švýcarsko MeSH
- Názvy látek
- antigeny CD274 MeSH
- CD274 protein, human MeSH Prohlížeč
- RAD51 protein, human MeSH Prohlížeč
- rekombinasa Rad51 MeSH
BACKGROUND: Regional variability in dural sinus (DS) wall thickness in posterior cranial fossa (PCF) have not been studied in detail yet. OBJECTIVE: To clarify the possible regional variability in DS wall thickness and determine the occurrence and localization of the chordae Willisii (CW) in PCF. METHODS: Fifty-nine human cadaveric DSs of PCF were investigated. A measurement of the DS walls/dura mater/CW thickness of parafin-embedded/hematoxylin-eosin stained axial sections was performed by using Cell Sens Science Imaging Software (Olympus Corporation, Tokyo, Japan). RESULTS: The osseus wall (OW) was the thickest one in the confluens sinuum (CS) and the thinnest one in the jugular bulb (JB) and sigmoid sinus (P < .05). The biggest differences between individual walls were observed in the JB where the superior wall was almost twice as thick as the OW. At the transverse-sigmoid junction, the thickness of the walls was comparable. In the CS and transverse sinuses, the OW was even thicker than the surrounding dura mater. The occurrence and thickness of the CW increased from the JB towards CS and prevailed on the right side. An overall number of the CW in PCF was comparable to that observed in the superior sagittal sinus. CONCLUSION: The present study displayed for the first time the regional variability in the DS walls thickness and occurrence of the CW in PCF. Application of these findings may afford greater freedom in exposure of the DSs or neoplasms adhering to the DSs.
- Klíčová slova
- Chordae willisii, Dural sinus, Posterior cranial fossa, Wall thickness,
- MeSH
- dospělí MeSH
- dura mater anatomie a histologie MeSH
- individualita MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mrtvola MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- sinus durae matris anatomie a histologie MeSH
- zadní jáma lební anatomie a histologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři 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.
- Klíčová slova
- Cutoff finder, Disease free survival, KM plotter, LC3A, Non-small cell lung cancer, Overall survival, Prognosis, Stone-like structures,
- 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
- Názvy látek
- MAP1LC3A protein, human MeSH Prohlížeč
- messenger RNA MeSH
- nádorové biomarkery MeSH
- proteiny asociované s mikrotubuly MeSH
BACKGROUND: Previously identified as a breast and ovarian cancer susceptibility gene, BRCA1 has gained major scientific interest as a potential prognostic and/or predictive marker for various tumors, including non-small-cell lung cancer (NSCLC), the leading cause of cancer related mortality worldwide. BRCA1 plays a central role in DNA damage response (DDR. It undergoes phosphorylation by various DDR kinases at different serine residues, of which ser1524 is known to be specifically phosphorylated by ATM in response to genotoxic stress. METHODS: We performed BRCA1 immunohistochemistry on several tissue microarrays (TMAs) of 113 early (I, II stage) and advanced (III, IV stage) NSCLCs, using MS110 antibody against the BRCA1 N-terminal and S1524 antibody against the phosphorylated form of BRCA1 protein at ser1524 (Abcam). Patients with III and IV stage disease were treated by adjuvant cisplatin-based chemotherapy. Staining results were correlated with overall survival (OS), disease free survival (DFS) and with the occurrence of brain metastases. RESULTS: BRCA1 S1524 nuclear positivity was significantly correlated with longer OS and DFS in stage I and II patients (P<0.05), while OS and DFS were shorter in S1524 positive stage III and IV patients (P<0.05). No significant correlation was found with brain metastases. CONCLUSION: The results show that BRCA1 phosphorylaton, at least in ser1524, differentiates the fate of early and advanced NSCLC as well as response to chemotherapy, but the underlying mechanisms are not completely understood. Detection of phosphorylated forms of BRCA1 might serve as a useful prognostic and predictive marker for patients with NSCLC.
- MeSH
- adjuvantní chemoterapie MeSH
- cisplatina terapeutické užití MeSH
- fosforylace fyziologie MeSH
- imunohistochemie MeSH
- kohortové studie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádorové biomarkery metabolismus MeSH
- nádory mozku farmakoterapie metabolismus sekundární MeSH
- nádory plic farmakoterapie metabolismus MeSH
- nemalobuněčný karcinom plic farmakoterapie metabolismus sekundární MeSH
- přežití bez známek nemoci MeSH
- prognóza MeSH
- protein BRCA1 metabolismus MeSH
- protinádorové látky terapeutické užití MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- BRCA1 protein, human MeSH Prohlížeč
- cisplatina MeSH
- nádorové biomarkery MeSH
- protein BRCA1 MeSH
- protinádorové látky MeSH
BACKGROUND: Barrett's esophagus (BE) is a disease with a rising prevalence in western countries probably due to the unhealthy lifestyle. In significant number of cases it develops to esophageal adenocarcinoma. Two decades ago, important gene regulators (microRNAs) were discovered and their attendance in the process of malignant transformation was demonstrated (e.g. miR-192, 196a, 203). Our aim was to select the patients with the increased risk of malignant transformation before the cancer develops. METHODS: 71 patients with BE disease were selected, slides from FFPE blocks were prepared, the lesions were microdissected and a qPCR relative expression analysis for selected microRNAs (generally known to be connected with malignant transformation process) was carried out. RESULTS: We demonstrated unequivocal statistically significant upregulation of two microRNAs (miR-192, 196a) and downregulation of miR-203 and positive miR-196a correlation with progression from intestinal metaplasia to adenocarcinoma compared to normal individuals. CONCLUSIONS: We hypothesize that there do exist changes of selected microRNAs which can undoubtedly distinguish the patients with BE from normal healthy individuals.
- MeSH
- adenokarcinom diagnóza etiologie genetika MeSH
- Barrettův syndrom komplikace diagnóza genetika MeSH
- biologické markery analýza MeSH
- down regulace genetika MeSH
- laserová záchytná mikrodisekce MeSH
- lidé MeSH
- metaplazie diagnóza genetika MeSH
- mikro RNA analýza genetika MeSH
- nádory jícnu diagnóza etiologie genetika MeSH
- progrese nemoci MeSH
- regulace genové exprese u nádorů genetika MeSH
- retrospektivní studie MeSH
- rizikové faktory MeSH
- střeva patologie MeSH
- upregulace genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biologické markery MeSH
- mikro RNA MeSH
- MIRN192 microRNA, human MeSH Prohlížeč
- MIRN196 microRNA, human MeSH Prohlížeč
- MIRN203 microRNA, human MeSH Prohlížeč
