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Commensal microbiota contribute to gut homeostasis by inducing transcription of mucosal genes. Analysis of the impact of various microbiota on intestinal tissue provides an important insight into the function of this organ. We used cDNA microarrays to determine the gene expression signature of mucosa isolated from the small intestine and colon of germ-free (GF) mice and animals monoassociated with two E. coli strains. The results were compared to the expression data obtained in conventionally reared (CR) mice. In addition, we analyzed gene expression in colon organoids derived from CR, GF, and monoassociated animals. The analysis revealed that the complete absence of intestinal microbiota mainly affected the mucosal immune system, which was not restored upon monoassociation. The most important expression changes observed in the colon mucosa indicated alterations in adipose tissue and lipid metabolism. In the comparison of differentially expressed genes in the mucosa or organoids obtained from GF and CR mice, only six genes were common for both types of samples. The results show that the increased expression of the angiopoietin-like 4 (Angptl4) gene encoding a secreted regulator of lipid metabolism indicates the GF status.
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Perfilação da Expressão Gênica , Vida Livre de Germes/genética , Mucosa Intestinal/metabolismo , Organoides/metabolismo , Animais , Biomarcadores/metabolismo , Colo/metabolismo , Escherichia coli/fisiologia , Regulação da Expressão Gênica , Sistema Imunitário/metabolismo , Imunidade nas Mucosas , Camundongos Endogâmicos BALB C , MicrobiotaRESUMO
Leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4) is produced in a broad spectrum of mouse embryonic and adult tissues and its deficiency results in embryonal or perinatal lethality. The LGR4 function was mainly related to its potentiation of canonical Wnt signaling; however, several recent studies associate LGR4 with additional signaling pathways. To obtain a suitable tool for studying the signaling properties of Lgr4, we generated a tagged variant of the Lgr4 receptor using gene targeting in the mouse oocyte. The modified Lgr4 allele expresses the Lgr4 protein fused with a triple hemagglutinin (3HA) tag located at the extracellular part of the protein. The allele is fully functional, enabling tracking of Lgr4 expression in the mouse tissues. We also show that via surface labeling, the 3HA tag allows direct isolation and analysis of living Lgr4-positive cells obtained from the small intestinal crypts. Finally, the HA tag-specific antibody can be employed to characterize the biochemical features of Lgr4 and to identify possible biding partners of the protein in cells derived from various mouse tissues.
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Marcação de Genes/métodos , Hemaglutininas/genética , Receptores Acoplados a Proteínas G/genética , Via de Sinalização Wnt/genética , Alelos , Animais , Feminino , Humanos , Camundongos , Oócitos/metabolismo , Gravidez , Ligação Proteica/genética , Distribuição TecidualRESUMO
BACKGROUND & AIMS: The Wnt signaling pathway is required for maintenance of the intestinal epithelia; blocking this pathway reduces the proliferative capacity of the intestinal stem cells. However, aberrant Wnt signaling leads to intestinal cancer. We investigated the roles of the Wnt pathway in homeostasis of the intestinal epithelium and during malignant transformation in human cells and mice. METHODS: We performed chromatin immunoprecipitation (ChIP) with DNA microarray analysis (ChIP-on-chip) to identify genes regulated by Wnt signaling in human colorectal cancer cells Colo320, DLD1, LS174T, and SW480. Formation of intestinal tumor was induced in C57BL/6J mice using azoxymethane and dextran sulfate. Intestinal tissues from these mice, as well as Apc(+/Min) and Apc(CKO/CKO)/Lgr5-EGFP-IRES-CreERT2 mice, were analyzed by immunohistochemistry and in situ hybridization. RESULTS: We identified promoter regions of 960 genes that interacted with the Wnt pathway nuclear effector T-cell factor 4 in 4 different human colorectal cancer-derived cell lines; 18 of these promoters were present in all chromatin precipitates. Wnt signaling up-regulated a member of the tumor necrosis factor receptor superfamily called TROY. Levels of TROY messenger RNA were increased in human cells with deficiencies in the adenomatous polyposis coli (APC) gene and in cells stimulated with the Wnt3a ligand. Expression of Troy was significantly up-regulated in neoplastic tissues from mice during intestinal tumorigenesis. Lineage tracing experiments revealed that Troy is produced specifically by fast-cycling intestinal stem cells. TROY associated with a unique marker of these cells, leucine-rich repeat-containing G-protein coupled receptor (LGR) 5. In organoids established from the intestinal crypts, Troy suppressed signaling mediated by R-spondin, a Wnt agonist. CONCLUSIONS: TROY is up-regulated in human colorectal cancer cell lines and in intestinal tumors in mice. It functions as a negative modulator of the Wnt pathway in LGR5-positive stem cells.
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Transformação Celular Neoplásica/genética , Regulação Neoplásica da Expressão Gênica , Mucosa Intestinal/metabolismo , Células-Tronco Neoplásicas/metabolismo , Receptores Acoplados a Proteínas G/fisiologia , Receptores do Fator de Necrose Tumoral/fisiologia , Via de Sinalização Wnt/fisiologia , Animais , Apoptose , Linhagem Celular Tumoral , Proliferação de Células , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Humanos , Imuno-Histoquímica , Hibridização In Situ , Mucosa Intestinal/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neoplasias Experimentais , Células-Tronco Neoplásicas/patologia , Análise de Sequência com Séries de Oligonucleotídeos , RNA Neoplásico/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
This study investigates the relationships between the hydration kinetics of waterglass-activated slag and the development of its physical-mechanical properties, as well as its color change. To modify the calorimetric response of alkali-activated slag, hexylene glycol was selected from various alcohols for in-depth experiments. In presence of hexylene glycol, the formation of initial reaction products was restricted to the slag surface, which drastically slowed down the further consumption of dissolved species and slag dissolution and consequently delayed the bulk hydration of the waterglass-activated slag by several days. This allowed to show that the corresponding calorimetric peak is directly related to the rapid evolution of the microstructure and physical-mechanical parameters and to the onset of a blue/green color change recorded as a time-lapse video. Workability loss was correlated with the first half of the second calorimetric peak, while the most rapid increase in strengths and autogenous shrinkage was related to the third calorimetric peak. Ultrasonic pulse velocity increased considerably during both the second and third calorimetric peak. Despite the modified morphology of the initial reaction products, the prolonged induction period, and the slightly reduced degree of hydration induced by hexylene glycol, the overall mechanism of alkaline activation remained unchanged in the long-term perspective. It was hypothesized that the main issue of the use of organic admixtures in alkali-activated systems is the destabilizing effect of these admixtures on soluble silicates introduced into the system with an activator.
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Protection of concrete against aggressive influences from the surrounding environment becomes an important step to increase its durability. Today, alkali silicate solutions are advantageously used as pore-blocking treatments that increase the hardness and impermeability of the concrete's surface layer. Among these chemical substances, known as concrete densifiers, lithium silicate solutions are growing in popularity. In the present study, the chemical composition of the lithium silicate densifiers is put into context with the properties of the newly created insoluble inorganic gel responsible for the micro-filling effect. Fourier-transform infrared spectroscopy was used as a key method to describe the structure of the formed gel. In this context, the gelation process was studied through the evolution of viscoelastic properties over time using oscillatory measurements. It was found that the gelation process is fundamentally controlled by the molar ratio of SiO2 and Li2O in the densifier. The low SiO2 to Li2O ratio promotes the gelling process, resulting in a rapidly formed gel structure that affects macro characteristics, such as water permeability, directly related to the durability of treated concretes.
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Biochar is a versatile carbon-rich organic material originating from pyrolyzed biomass residues that possess the potential to stabilize organic carbon in the soil, improve soil fertility and water retention, and enhance plant growth. For the utilization of biochar as a soil conditioner, the mutual interconnection of the physicochemical properties of biochar with the production conditions used during the pyrolysis (temperature, heating rate, residence time) and the role of the origin of used biomass seem to be crucial. The aim of the research was focused on a comparison of the properties of biochar samples (originated from oat brans, mixed woodcut, corn residues and commercial compost) produced at different temperatures (400-700 °C) and different residence times (10 and 60 min). The results indicated similar structural features of produced biochar samples; nevertheless, the original biomass showed differences in physicochemical properties. The morphological and structural analysis showed well-developed aromatic porous structures for biochar samples originated from oat brans, mixed woodcut and corn residues. The higher pyrolysis temperature resulted in lower yields; however, it provided products with higher content of organic carbon and a more developed surface area. The lignocellulose biomass with higher contents of lignin is an attractive feedstock material for the production of biochar with potential agricultural applications.
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Trophoblastic cell surface antigen 2 (TROP2) is a membrane glycoprotein overexpressed in many solid tumors with a poor prognosis, including intestinal neoplasms. In our study, we show that TROP2 is expressed in preneoplastic lesions, and its expression is maintained in most colorectal cancers (CRC). High TROP2 positivity correlated with lymph node metastases and poor tumor differentiation and was a negative prognostic factor. To investigate the role of TROP2 in intestinal tumors, we analyzed two mouse models with conditional disruption of the adenomatous polyposis coli (Apc) tumor-suppressor gene, human adenocarcinoma samples, patient-derived organoids, and TROP2-deficient tumor cells. We found that Trop2 is produced early after Apc inactivation and its expression is associated with the transcription of genes involved in epithelial-mesenchymal transition, the regulation of migration, invasiveness, and extracellular matrix remodeling. A functionally similar group of genes was also enriched in TROP2-positive cells from human CRC samples. To decipher the driving mechanism of TROP2 expression, we analyzed its promoter. In human cells, this promoter was activated by ß-catenin and additionally by the Yes1-associated transcriptional regulator (YAP). The regulation of TROP2 expression by active YAP was verified by YAP knockdown in CRC cells. Our results suggest a possible link between aberrantly activated Wnt/ß-catenin signaling, YAP, and TROP2 expression.
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A large proportion of colorectal carcinomas (CRC) evolve from colorectal adenomas. However, not all individuals with colonic adenomas have a risk of CRC substantially higher than those of the general population. The aim of the study was to determine the differences or similarities of mutation profile among low- and high-grade adenomas and in situ carcinoma with detailed follow up. We have investigated the mutation spectrum of well-known genes involved in CRC (such as APC, BRAF, EGFR, NRAS, KRAS, PIK3CA, POLE, POLD1, SMAD4, PTEN, and TP53) in a large, well-defined series of 96 adenomas and in situ carcinomas using a high-throughput genotyping technique. Besides, the microsatellite instability and APC and MLH1 promoter methylation were studied as well. We observed a high frequency of pathogenic variants in the studied genes. The APC, KRAS and TP53 mutation frequencies were slightly lower in adenoma samples than in in situ carcinoma samples. Further, when we stratified mutation frequency based on the grade, the frequency distribution was as follows: low-grade adenoma-high-grade adenomas-in situ carcinoma: APC gene 42.9-56.0-54.5%; KRAS gene 32.7-32.0-45.5%; TP53 gene 8.2-20.0-18.2%. The occurrence of KRAS mutation was associated with the presence of villous histology and methylation of the APC promoter was significantly associated with the presence of POLE genetic variations. However, no association was noticed with the presence of any singular mutation and occurrence of subsequent adenoma or CRC. Our data supports the multistep model of gradual accumulation of mutations, especially in the driver genes, such as APC, TP53 and KRAS.
Assuntos
Adenoma/patologia , Biomarcadores Tumorais/genética , Carcinoma in Situ/patologia , Neoplasias Colorretais/patologia , Instabilidade de Microssatélites , Mutação , Adenoma/genética , Idoso , Carcinoma in Situ/genética , Neoplasias Colorretais/genética , Metilação de DNA , Feminino , Seguimentos , Humanos , Masculino , PrognósticoRESUMO
Elevated levels of survivin, telomerase catalytic subunit (TERT), integrin-linked kinase (ILK), cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS) and the regulatory factors c-MYB and Tcf-4 are often found in human cancers including colorectal cancer (CRC) and have been implicated in the development and progression of tumorigenesis. The aim of this study was to determine the expression of these genes in mouse models of sporadic and colitis-associated CRC. To address these issues, we used qRT-PCR approach to determine changes in gene expression patterns of neoplastic cells (high-grade dysplasia/intramucosal carcinoma) and surrounding normal epithelial cells in A/J and ICR mouse strains using laser microdissection. Both strains were injected with azoxymethane and ICR mice were also given drinking water that contained 2% dextran sodium sulphate. In both sporadic (A/J mice) and colitis-associated (ICR mice) models of CRC, the levels of TERT mRNA, COX-2 mRNA and Tcf-4 mRNA were higher in neoplastic cells than in surrounding normal epithelial cells. In contrast, survivin mRNA was upregulated only in neoplastic cells from A/J mice and ILK mRNA was upregulated only in neoplastic cells from ICR mice. However, the expression of iNOS mRNA was similar in normal and neoplastic cells in both models and c-MYB mRNA was actually downregulated in neoplastic cells compared with normal cells in both models. These findings suggest that the genetic background and/or the molecular mechanisms of tumorigenesis associated with genotoxic insults and colonic inflammation influence the gene expression of mTERT, COX-2, Tcf-4, c-MYB, ILK and survivin in colon epithelial neoplasia.
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Proteínas Reguladoras de Apoptose/genética , Apoptose/genética , Proteínas de Ciclo Celular/genética , Proliferação de Células , Colite/genética , Neoplasias do Colo/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Animais , Azoximetano , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Transformação Celular Neoplásica/induzido quimicamente , Transformação Celular Neoplásica/genética , Colite/induzido quimicamente , Colite/complicações , Colite/patologia , Neoplasias do Colo/induzido quimicamente , Neoplasias do Colo/patologia , Ciclo-Oxigenase 2/genética , Sulfato de Dextrana , Modelos Animais de Doenças , Progressão da Doença , Perfilação da Expressão Gênica/métodos , Proteínas Inibidoras de Apoptose , Masculino , Camundongos , Camundongos Endogâmicos ICR , Microdissecção , Proteínas Associadas aos Microtúbulos/genética , Óxido Nítrico Sintase Tipo II/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas c-myb/genética , Proteínas Repressoras , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Survivina , Telomerase/genética , Fator de Transcrição 4RESUMO
Increasing utilization of secondary raw materials and alternative fuels results in increasing contents of metals in cements. Zinc is one of these elements. It comes to cement with secondary raw materials such as slag or fly ash or by the utilization of used tires as an alternative fuel. Zinc ions significantly prolong the hydration process in cement. This work deals with the influence of zinc ions in the form of very poorly soluble ZnO salt and easily soluble ZnCl2 and Zn(NO3)2 on the hydration of cement blended with fly ash. Zinc was dosed in the range of 0.05%, 0.1%, 0.5% and 1% of cement weight. The effect of zinc on hydration was monitored by isothermal and isoperibolic calorimetry. A 15% addition of fly ash to cement mainly causes further retardation of hydration reactions due to the reactions of fly ash particles with Ca2+ ions from cement. The strongest effect on the hydration retardation from all investigated compounds showed in ZnO as it dissolves very slowly. On the contrary, for the dosage of 1% of zinc in the form of ZnCl2 significant acceleration of hydration occurred. In this work, a synergistic effect on the prolongation of hydration with a combination of cement, zinc and fly ash was demonstrated. The lengths of induction periods were assessed from detected calorimetric curves and from these lengths the curves were gained by fitting with the exponential function. Final products were next analyzed using X-ray diffraction.
Assuntos
Anticorpos Monoclonais Humanizados , Melanoma , Compostos de Fenilureia , Quinolinas , Neoplasias Cutâneas , Humanos , Melanoma/tratamento farmacológico , Melanoma/secundário , Melanoma/patologia , Anticorpos Monoclonais Humanizados/efeitos adversos , Anticorpos Monoclonais Humanizados/uso terapêutico , Anticorpos Monoclonais Humanizados/administração & dosagem , Compostos de Fenilureia/administração & dosagem , Quinolinas/uso terapêutico , Quinolinas/administração & dosagem , Quinolinas/efeitos adversos , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/patologia , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , AdultoRESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
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The first step in the development of human colorectal cancer is aberrant activation of the Wnt signaling pathway. Wnt signaling hyperactivation is predominantly caused by loss-of-function mutations in the adenomatous polyposis coli (APC) gene that encodes the pathway negative regulator. In order to identify genes affected by the Apc loss, we performed expression profiling of intestinal epithelium isolated from mice harboring a conditional Apc allele. The gene encoding transcriptional factor msh homeobox 1 (Msx1) displayed robust upregulation upon Apc inactivation. Histological analysis of the Apc-deficient epithelium revealed that in the small intestine, the Msx1 protein was localized exclusively in ectopic crypts, i.e., in pockets of proliferating cells abnormally positioned on the villi. Ablation of the Msx1 gene leads to the disappearance of ectopic crypts and loss of differentiated cells. Moreover, tumors arising from Msx1-deficient cells display altered morphology reminiscent of villous adenomas. In human tumor specimens, MSX1 displayed significantly increased expression in colonic neoplasia with a descending tendency during the lesion progression towards colorectal carcinoma. In summary, the results indicate that Msx1 represents a novel marker of intestinal tumorigenesis. In addition, we described the previously unknown relationship between the Msx1-dependent formation of ectopic crypts and cell differentiation.
Assuntos
Proteína da Polipose Adenomatosa do Colo/genética , Mucosa Intestinal/patologia , Intestino Delgado/patologia , Fator de Transcrição MSX1/genética , Fator de Transcrição MSX1/metabolismo , Proteína da Polipose Adenomatosa do Colo/metabolismo , Animais , Diferenciação Celular , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Neoplasias Colorretais/genética , Neoplasias Colorretais/patologia , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Mucosa Intestinal/metabolismo , Camundongos Knockout , Via de Sinalização Wnt , beta Catenina/metabolismoRESUMO
Radiation and chemotherapy represent standard-of-care cancer treatments. However, most patients eventually experience tumour recurrence, treatment failure and metastatic dissemination with fatal consequences. To elucidate the molecular mechanisms of resistance to radio- and chemotherapy, we exposed human cancer cell lines (HeLa, MCF-7 and DU145) to clinically relevant doses of 5-azacytidine or ionizing radiation and compared the transcript profiles of all surviving cell subpopulations, including low-adherent stem-like cells. Stress-mobilized low-adherent cell fractions differed from other survivors in terms of deregulation of hundreds of genes, including those involved in interferon response. Exposure of cancer cells to interferon-gamma but not interferon-beta resulted in the development of a heterogeneous, low-adherent fraction comprising not only apoptotic/necrotic cells but also live cells exhibiting active Notch signalling and expressing stem-cell markers. Chemical inhibition of mitogen-activated protein kinase/ERK kinase (MEK) or siRNA-mediated knockdown of extracellular signal-regulated kinase 1/2 (Erk1/2) and interferon responsible factor 1 (IRF1) prevented mobilization of the surviving low-adherent population, indicating that interferon-gamma-mediated loss of adhesion and anoikis resistance required an active Erk pathway interlinked with interferon signalling by transcription factor IRF1. Notably, a skin-specific protein suprabasin (SBSN), a recently identified oncoprotein, was among the top scoring genes upregulated in surviving low-adherent cancer cells induced by 5-azacytidine or irradiation. SBSN expression required the activity of the MEK/Erk pathway, and siRNA-mediated knockdown of SBSN suppressed the low-adherent fraction in irradiated, interferon-gamma- and 5-azacytidine-treated cells, respectively, implicating SBSN in genotoxic stress-induced phenotypic plasticity and stress resistance. Importantly, SBSN expression was observed in human clinical specimens of colon and ovarian carcinomas, as well as in circulating tumour cells and metastases of the 4T1 mouse model. The association of SBSN expression with progressive stages of cancer development indicates its role in cancer evolution and therapy resistance.
Assuntos
Antígenos de Diferenciação/genética , Antineoplásicos/farmacologia , Azacitidina/farmacologia , Interferons/farmacologia , Proteínas de Neoplasias/genética , Neoplasias/tratamento farmacológico , Animais , Anoikis/efeitos dos fármacos , Anoikis/efeitos da radiação , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Neoplasias/genética , Neoplasias/radioterapia , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/efeitos da radiação , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/efeitos da radiaçãoRESUMO
Neoplastic growth is frequently associated with genomic DNA methylation that causes transcriptional silencing of tumor suppressor genes. We used a collection of colorectal polyps and carcinomas in combination with bioinformatics analysis of large datasets to study the expression and methylation of Hypermethylated in cancer 1 (HIC1), a tumor suppressor gene inactivated in many neoplasms. In premalignant stages, HIC1 expression was decreased, and the decrease was linked to methylation of a specific region in the HIC1 locus. However, in carcinomas, the HIC1 expression was variable and, in some specimens, comparable to healthy tissue. Importantly, high HIC1 production distinguished a specific type of chemotherapy-responsive tumors.
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The activity of the Wnt pathway undergoes complex regulation to ensure proper functioning of this principal signaling mechanism during development of adult tissues. The regulation may occur at several levels and includes both positive and negative feedback loops. In the present study we employed one of such negative feedback regulators, naked cuticle homolog 1 (Nkd1), to follow the Wnt pathway activity in the intestine and liver and in neoplasia originated in these organs. Using lineage tracing in transgenic mice we localized Nkd1 mRNA to the bottom parts of the small intestinal crypts and hepatocytes surrounding the central vein of the hepatic lobule. Furthermore, in two mouse models of intestinal tumorigenesis, Nkd1 expression levels were elevated in tumors when compared to healthy tissue. We utilized a collection of human intestinal polyps and carcinomas to confirm that NKD1 represents a robust marker of neoplastic growth. In addition, expression analysis of NKD1 in liver cancer showed that high expression levels of the gene distinguish a subclass of hepatocellular carcinomas related to aberrant Wnt signaling. Finally, our results were confirmed by bioinformatic analysis of large publicly available datasets that included gene expression profiling and high-throughput sequencing data of human colon and liver cancer specimens.
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Carcinoma Hepatocelular/patologia , Proteínas de Transporte/metabolismo , Neoplasias Intestinais/patologia , Neoplasias Hepáticas/patologia , Proteínas Wnt/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Proteína da Polipose Adenomatosa do Colo/deficiência , Proteína da Polipose Adenomatosa do Colo/genética , Proteína da Polipose Adenomatosa do Colo/metabolismo , Animais , Proteínas de Ligação ao Cálcio , Carcinoma Hepatocelular/metabolismo , Proteínas de Transporte/genética , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Hepatócitos/metabolismo , Hepatócitos/patologia , Humanos , Neoplasias Intestinais/metabolismo , Neoplasias Hepáticas/metabolismo , Camundongos , Camundongos Transgênicos , Mutação , RNA Mensageiro/metabolismo , Transdução de Sinais , Transcrição Gênica , beta Catenina/genética , beta Catenina/metabolismoRESUMO
Muir-Torre syndrome (MTS), a rare variant of the hereditary non polyposis colorectal cancer syndrome, is an autosomal dominant genodermatosis characterised by coincidence of sebaceous gland neoplasms (sebaceous adenoma, epithelioma, or carcinoma) and at least one internal malignancy. The underlying cause of MTS is a germline mutation in DNA mismatch repair genes MSH2, MLH1 and MSH6. We report the case of a 52-year-old caucasian woman with the development of metachronous colon cancer at the age of 38 years, uterine cancer at the age of 43 years, and unique occurrence of synchronous gastric and sebaceous carcinomas related to germline point mutation c. 2194A>T in the last exon of MLH1 gene, resulting in truncated protein in C-terminal region p. Lys732X due to premature stop codon. This mutation, not previously reported in MTS, disrupts the function of MutL complexes presumably by preventing the interaction with PMS1/PMS2 and impairing the endonuclease active site. This case points out the importance of sebaceous neoplasia, especially sebaceous adenocarcinoma, as cutaneous markers of MTS for timely implementation of cancer screening programs.
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Proteínas Adaptadoras de Transdução de Sinal/genética , Adenocarcinoma Sebáceo/genética , Síndrome de Muir-Torre/genética , Neoplasias Primárias Múltiplas/genética , Proteínas Nucleares/genética , Neoplasias das Glândulas Sebáceas/genética , Neoplasias Gástricas/genética , Adenocarcinoma Sebáceo/patologia , Feminino , Mutação em Linhagem Germinativa , Humanos , Pessoa de Meia-Idade , Síndrome de Muir-Torre/patologia , Proteína 1 Homóloga a MutL , Neoplasias Primárias Múltiplas/patologia , Mutação Puntual , Neoplasias das Glândulas Sebáceas/patologia , Neoplasias Gástricas/patologiaRESUMO
Although the effects of glucocorticoids on proliferation, differentiation and apoptosis are well known, and steroid hormones have been identified to play a role in pathogenesis and the development of various cancers, limited data are available regarding the relationship between the local metabolism of glucocorticoids and colorectal adenocarcinoma (CRC) formation. Glucocorticoid metabolism is determined by 11ß-hydroxysteroid dehydrogenases type 1 and 2 (11HSD1, 11HSD2), which increase the local concentration of cortisol due to the reduction of cortisone, or decrease this concentration due to the oxidation of cortisol. The objective of this study was to evaluate the extent of 11HSD1 and 11HSD2 mRNA in pre-malignant colorectal polyps and in CRC. The specimens were retrieved from patients by endoscopic or surgical resection and the expression of 11HSD1 and 11HSD2 was measured by real-time PCR. The polyps were of the following histological types: hyperplastic polyps and adenomas with low- or high-grade dysplasia. The neoplastic tissue of CRC obtained during tumor surgery was also studied. It was found that 11HSD2 was not only downregulated in CRC but already in the early stages of neoplastic transformation (adenoma with low-grade dysplasia). In contrast, the level of 11HSD1 was significantly increased in CRC but not in pre-malignant polyps. The results demonstrate that the downregulation of 11HSD2 gene expression is a typical feature of the development of colorectal polypous lesions and their transformation into CRC.