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Lack of expression of the tumour suppressor gene caudal-type homeobox 2 (CDX2) associates with poor outcomes in early stage colorectal cancer (CRC). Yet its prognostic value in the context of other prognostic biomarkers in metastatic CRC (mCRC) is unknown. Overexpressed cyclooxygenase-2 (COX2) has been reported in advanced CRC. However, CDX2 and COX2 relationship in mCRC remains undetermined. We aimed to assess their expression in mCRC tumours from a clinically characterised cohort and their influence on overall survival (OS) and progression-free survival (PFS) in first line. Among 720 consecutive mCRC patients, 346 had tumour samples appropriate for tissue microarray assembly and immunohistochemistry analyses. Clinical and survival data were retrospectively assessed. Loss of CDX2 expression was detected in 27 (7.8%) samples, enriched in poorly differentiated tumours (20%; p < 0.01) and in those with the BRAF p.V600E variant (40%; p < 0.01). Most tumours (93.4%) expressed COX2. COX2-negative samples were enriched in poorly differentiated mCRC. In unadjusted analyses, median OS (p < 0.001) and median PFS (p < 0.05) were inferior for patients with CDX2-negative versus CDX2-positive tumours. In conclusion, loss of CDX2 was significantly associated with poorly differentiated mCRC and BRAF p.V600E allele and a prognostic marker of worse OS.
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Oocyte activation via dual inhibition of protein synthesis and phosphorylation has improved in vitro embryo production in different mammalian species. In this study, we evaluated the effects of the combination of cycloheximide (CHX), dimethyl amino purine (DMAP), and anisomycin (ANY) on the activation of bovine oocytes, particularly on dynamics of MPF and MAPKs, embryonic developmental potential, and quality. The results showed that the cleavage and blastocyst rates, as well as levels of CCNB1, CDK1, p-CDK1Thr161, and p-CDK1Thr14-Tyr15, were similar among groups; ANY and ANY + CHX reduced the expression of ERK1/2 compared to DMAP-combinations (p < 0.05), whereas ANY + DMAP, CHX + DMAP, and ANY + CHX + DMAP reduced p-ERK1/2 compared to ANY and ANY + CHX treatments (p < 0.05). The quality of blastocysts in terms of cell counts, their allocation, and the numbers of TUNEL-positive cells did not differ among groups. However, transcript levels of POU5F1 were higher in embryos derived from ANY + CHX + DMAP treatment compared to other groups, while expression levels of CDX2 did not show differences. In addition, the BCL2A1/BAX ratio of the ANY + CHX + DMAP treatment was significantly low compared to the ANY treatment (p < 0.05) and did not differ significantly from the other treatments. In conclusion, oocyte activation by dual inhibition of protein synthesis and phosphorylation induces MPF inactivation without degradation of CCNB1, while MAPK inactivation occurs differentially between these inhibitors. Thus, although the combined use of these inhibitors does not affect early developmental competence in vitro, it positively impacts the expression of transcripts associated with embryonic quality.
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Fator Promotor de Maturação , Partenogênese , Bovinos , Animais , Proteínas Quinases Ativadas por Mitógeno , Adenina/farmacologia , Oócitos , Cicloeximida/farmacologia , Blastocisto , Anisomicina/farmacologia , MamíferosRESUMO
Background: Colorectal cancer is the most frequent gastrointestinal malignancy worldwide. The value of adjuvant treatment is controversial in Stages I and II. Objective: The aim of this study was to construct post-operative prognostic models applicable to patients with stages I-II colon carcinoma (CC). Methods: This is a retrospective cohort study of patients with Stage I-II CC treated over a 25-year period. Exposure was defined as clinical, histopathological, and immunohistochemical factors (including CDX2 and MUC2 expression). Patients were randomly allocated to either a "modeling set" or a "validation set". Factors associated with recurrence, disease-free survival (DFS), and overall survival (OS) were defined in the "modeling set". Their performances were tested in the "validation set". Results: From a total of 556 recruited patients, 339 (61%) were allocated to the "modeling set" and 217 (39%) to the "validation set". Three models explaining recurrence, DFS, and OS were described. Tumor location in the left colon (Hazards ratio [HR] = 1.57; 95% Confidence interval [CI] 0.99-2.48), lymphocyte (HR = 0.46; 96% CI 0.27-0.88) and monocyte (HR = 0.99; 95% CI 0.99-1) counts, neutrophil/platelet ratio (HR = 1.3; 95% CI 0.74-2.3, and HR = 2.3; 95% CI 1.3-4.1; for second and third category, respectively), albumin/monocyte ratio (HR = 0.43; 95% CI 0.21-0.87), and microscopic residual disease after surgery (HR = 8.7; 95% CI 3.1-24) were independently associated with OS. T classification and expression of CDX2 and/or MUC2 were not independently associated with recurrence or prognosis. Conclusion: These models are simple and readily available, and distinguish the risk and prognosis in patients with CC stages I and II; these models require cheaper processes than the use of more sophisticated molecular biology techniques. They may guide either the need for adjuvant therapy versus post-operative surveillance only, as well as aid in the design of clinical trials.
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Carcinoma , Neoplasias do Colo , Humanos , Prognóstico , Estudos Retrospectivos , Neoplasias do Colo/cirurgia , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/patologia , Carcinoma/patologia , Estadiamento de NeoplasiasRESUMO
ABSTRACT Background: Colorectal cancer is the most frequent gastrointestinal malignancy worldwide. The value of adjuvant treatment is controversial in Stages I and II. Objective: The aim of this study was to construct post-operative prognostic models applicable to patients with stages I-II colon carcinoma (CC). Methods: This is a retrospective cohort study of patients with Stage I-II CC treated over a 25-year period. Exposure was defined as clinical, histopathological, and immunohistochemical factors (including CDX2 and MUC2 expression). Patients were randomly allocated to either a "modeling set" or a "validation set". Factors associated with recurrence, disease-free survival (DFS), and overall survival (OS) were defined in the "modeling set". Their performances were tested in the "validation set". Results: From a total of 556 recruited patients, 339 (61%) were allocated to the "modeling set" and 217 (39%) to the "validation set". Three models explaining recurrence, DFS, and OS were described. Tumor location in the left colon (Hazards ratio [HR] = 1.57; 95% Confidence interval [CI] 0.99-2.48), lymphocyte (HR = 0.46; 96% CI 0.27-0.88) and monocyte (HR = 0.99; 95% CI 0.99-1) counts, neutrophil/platelet ratio (HR = 1.3; 95% CI 0.74-2.3, and HR = 2.3; 95% CI 1.3-4.1; for second and third category, respectively), albumin/monocyte ratio (HR = 0.43; 95% CI 0.21-0.87), and microscopic residual disease after surgery (HR = 8.7; 95% CI 3.1-24) were independently associated with OS. T classification and expression of CDX2 and/or MUC2 were not independently associated with recurrence or prognosis. Conclusion: These models are simple and readily available, and distinguish the risk and prognosis in patients with CC stages I and II; these models require cheaper processes than the use of more sophisticated molecular biology techniques. They may guide either the need for adjuvant therapy versus post-operative surveillance only, as well as aid in the design of clinical trials.
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Siendo el cáncer gástrico la 3ª causa de muerte por cáncer en Chile, y existiendo estrategias de tamizaje consistentes en pesquisa de lesiones preneoplásicas de la mucosa gástrica, es relevante conocer los aspectos genéticos y moleculares que puedan ser aplicados, en la optimización de dichas estrategias a grupos de mayor riesgo. El objetivo de este manuscrito fue revisar la evidencia actual en los aspectos señalados, y de la inmunohistoquímica de 4 marcadores (p53, CDX2, MUC2 y S100A9) en la mucosa gástrica normal y en las lesiones preneoplásicas de la misma.
SUMMARY: Since gastric cancer is the 3rd leading cause of death from cancer in Chile, and there are screening strategies consisting of screening for preneoplastic lesions of the gastric mucosa, it is important to know certain genetic and molecular aspects that can be applied in optimizing these strategies for higher risk groups. The aim of this manuscript was to review the current evidence on the aforementioned aspects, and on the immunohistochemistry of 4 markers (p53, CDX2, MUC2 and S100A9) in normal gastric mucosa and in its preneoplastic lesions.
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Humanos , Lesões Pré-Cancerosas/patologia , Neoplasias Gástricas/patologia , Mucosa Gástrica/patologia , Lesões Pré-Cancerosas/genética , Lesões Pré-Cancerosas/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Imuno-Histoquímica , Biomarcadores Tumorais , Programas de Rastreamento , Fatores de Risco , Genes p53 , Mucina-2 , Fator de Transcrição CDX2 , Mucosa Gástrica/metabolismo , MetaplasiaRESUMO
Supplementing embryonic culture medium with fetal bovine serum (FBS) renders this medium undefined. Glucose and growth factors present in FBS may affect the results of cell differentiation studies. This study tested the hypothesis that FBS supplementation during in vitro culture (IVC) alters cell differentiation in early bovine embryo development. Bovine embryos were produced in vitro and randomly distributed into three experimental groups at 90 h post insemination (90 hpi): the KSOM-FBS group, which consisted of a 5% (v/v) FBS supplementation; the KSOM33 group, with the renewal of 33% of medium volume; and the KSOM-Zero group, without FBS supplementation nor renewal of the culture medium. The results showed that the blastocyst rate (blastocyst/oocytes) at 210 hpi in the KSOM-FBS group was higher than in the KSOM-Zero group but not different from the KSOM33 group. There were no significant changes in metabolism-related aspects, such as fluorescence intensities of CellROX Green and MitoTracker Red or reduced nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD+). Immunofluorescence analysis of CDX2 revealed that the lack of FBS or medium supplementation reduced the number of trophectoderm (TE) cells and total cells. Immunofluorescence analysis revealed a reduction of SOX17-positive cell numbers after FBS supplementation compared with the KSOM33 group. Therefore, we concluded that FBS absence reduced blastocyst rates; however, no reduction occurred when there was a 33% volume renewal of the medium at 90 hpi. We also concluded that FBS supplementation altered TE and primitive endoderm cell allocation during early bovine embryo development.
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Fertilização in vitro , Soroalbumina Bovina , Endoderma , Técnicas de Cultura Embrionária/métodos , Desenvolvimento Embrionário , Blastocisto , Meios de Cultura/farmacologiaRESUMO
A perda de expressão do gene supressor de tumores CDX2 (fator de transcrição homeótico tipo caudal 2) está associada a resultados desfavoráveis em câncer colorretal em estágio inicial. No entanto, o seu valor prognóstico no contexto de outros biomarcadores prognósticos em câncer colorretal metastático (CCRm) é desconhecido. Superexpressão da proteína ciclooxigenase-2 (COX2) foi relatada em câncer colorretal avançado. No entanto, a relação entre CDX2 e COX2 em CCRm permanece indeterminada. Nosso objetivo foi avaliar a sua expressão em tumores de CCRm de uma coorte clinicamente caracterizada bem como o seu impacto na sobrevida global (SG) e na sobrevida livre de progressão (SLP) na primeira linha de tratamento.Dentre 720 pacientes consecutivos com CCRm, 346 apresentavam amostras tumorais apropriadas para montagem de microarranjos de tecidos e análises de imuno-histoquímica. Dados clínicos e de sobrevida foram avaliados retrospectivamente. A perda de expressão de CDX2 foi detectada em 27 (7,8%) amostras, enriquecidas em tumores pouco diferenciados (20%; p<0,01) e naqueles com a variante BRAF p.V600E (40%; p<0,01). A maioria dos tumores (93,4%) expressou COX2. Amostras negativas para COX2 foram mais comuns em CCRm pouco diferenciados. Em análises não ajustadas, a mediana da SG (p<0,001) e a mediana da SLP (p<0,05) foram inferiores para pacientes com tumores CDX2-negativos em comparação com tumores CDX2-positivos. Em conclusão, a perda de CDX2 mostrou uma associação significativa com CCRm pouco diferenciado e o alelo BRAF p.V600E, sendo um marcador prognóstico de piora da sobrevida global.
INTRODUCTION: Lack of expression of the tumor suppressor gene CDX2 associates with poor outcomes in early stage colorectal cancer. Yet its prognostic value in the context of other prognostic biomarkers in metastatic CCR (CCRm) is unknown. Overexpressed cyclooxygenase-2 (COX2), encoded by the prostaglandin-endoperoxide synthase 2 gene has been reported in advanced CCR. However, CDX2 and COX2 relationship in CCRm remains undetermined. We aimed to assess their expression in CCRm tumors from a clinically characterized cohort and their influence on overall survival (OS) and progression-free survival (PFS) in first line. METHODS: Demographic and clinical data from mCRC were retrospectively analyzed. Appropriate tumor samples were collected for tissue microarray and analyzed by immunohistochemistry. RESULTS: Three hundred and forty six mCRC were included. Loss of CDX2 expression was detected in 27 (7.8%) samples, enriched in poorly differentiated tumors (20%; p<0.01) and in those with the BRAF p.V600E variant (40%; p<0.01). Most tumors (93.4%) expressed COX2. COX2-negative samples were enriched in poorly differentiated CCRm. In unadjusted analyses, median OS (p<0.001) and median PFS (p<0.05) were inferior for patients with CDX2-negative versus CDX2-positive tumors. CONCLUSION:Loss of CDX2 was significantly associated with poorly differentiated CCRm and BRAF p.V600E allele and a prognostic marker of worse OS.
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HumanosRESUMO
SUMMARY OBJECTIVE: Thyroid neoplasm incidence has increased worldwide, mostly due to the advancements in medical imaging and screening rates. The aberrant Wnt/β-catenin pathway has been identified as a key mechanism, and it has also been related to the metastatic activity of differentiated thyroid cancer. We aimed to verify the difference in the expression of Wnt3a, a canonical activator of the β-catenin signaling, and CDX-2, a transcription factor upregulated by Wnt/β-catenin pathway, in multinodular goiter and differentiated thyroid cancer and to determine their prognostic value. METHODS: We included 194 thyroid tissue surgical specimen and their clinicopathological data: study group (differentiated thyroid cancer, n=154) and control group (multinodular goiter, n=40). Immunohistochemistry (IHC) was performed on formalin-fixed, paraffin-embedded tissue by the primary antibodies Wnt3a and CDX-2. RESULTS: High Wnt3a expression was significantly associated with differentiated thyroid cancer (p=0.031). CDX-2 was negative in all differentiated thyroid cancer cases (100%) and also in multinodular goiter. Wnt3a expression was significantly associated with tumors ≤20 mm (p=0.044) and with the absence of capsule invasion (p=0.031). The multivariate analyses suggested that older age (≥55), independent of capsular invasion and tumor size, was an independent prognostic factor for Wnt3a expression (p=0.058). CONCLUSIONS: Wnt3a expression but not CDX-2 is correlated with differentiated thyroid cancer samples in comparison to multinodular goiter. Although its prognostic value was limited to tumor size and capsule invasion, a combined model in a panel of immune markers can add accuracy in the classification of challenging thyroid follicular-derived lesions.
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Introdução: A proteína CDX2 é um fator de transcrição específico do intestino, que está presente no tecido gástrico apenas quando há uma metaplasia intestinal. A metaplasia intestinal é uma lesão precursora do adenocarcinoma gástrico. O Ki67 é um biomarcador de proliferação celular. Objetivo: Verificar a presença da proteína CDX2 no adenocarcinoma gástrico. Comparar a expressão da CDX2 entre os diferentes graus de diferenciação e entre os níveis de proliferação celular. Método: A partir de 62 blocos histológicos contendo amostras de adenocarcinoma gástricos (4 bem diferenciados, 30 moderadamente diferenciados e 28 pouco diferenciados), foi feita a construção de blocos multiamostrais (TMA). Procedeu-se a marcação imunoistoquímica com os anticorpos escolhidos e realizou-se a leitura da área positiva imunocoradas. Resultados: 31 amostras foram positivas para a CDX2 e 31 negativas, sem diferença significativa entres os graus de diferenciação (p = 0,576). 38 amostras foram classificadas como de baixo grau de proliferação celular e 24 como de alto grau. Não houve diferença estatística de grau de proliferação celular entre os graus de diferenciação (p = 0,676). O grau de proliferação celular variou dependendo da expressão da CDX2 (p = 0,036). Conclusão: A expressão da proteína CDX2 esteve presente em 50% dos adenocarcinomas gástricos. Não houve diferença estatística da expressão do CDX2 entres os graus de diferenciação do adenocarcinoma gástrico. A proliferação celular variou dependendo da expressão da CDX2, havendo um maior nível de proliferação celular nas amostras que apresentaram expressão positiva para CDX2
Background: CDX2 protein is an intestinal specific transcription factor that is present in the gastric tissue only when there is intestinal metaplasia. Intestinal metaplasia is a gastric adenocarcinoma precursor injury. Ki67 is a cell proliferation biomarker. Objective: Verify the presence of CDX2 protein in gastric adenocarcinoma. Compare the CDX2 expression between the differentiation degree groups and between the cell proliferation degrees. Methods: It was collected 62 paraffin blocks containing the gastric adenocarcinoma samples (4 well differentiated adenocarcinoma, 30 moderately differentiated and 28 poorly differentiated). It was made the tissue microarrays blocks (TMA), so it was proceeded to immunohistochemical staining with the anti-CDX2 and the anti-Ki67 antibodies and it was accomplished the read of the positive immune-stained area. Results: 31 samples were positive for CDX2 expression, and 31 were negative. There was no statistical difference of the CDX2 expression between the differentiation degree groups (p = 0.576). 38 samples were classified as low degree of cell proliferation and 24 as high degree. There was no statistical difference of the cell proliferation degree between the differentiation degree groups (p = 0.676). The degree of cell proliferation ranged depending on the CDX2 expression (p = 0.036). Conclusion: Protein CDX2 expression was observed in 50% of gastric adenocarcinoma samples. There was no statistical difference of the CDX2 expression between the differentiation degree groups. The cell proliferation ranged depending of CDX2 expression, with a higher index of cell proliferation in the positive CDX2 expression samples
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Humanos , Neoplasias Gástricas , Imuno-Histoquímica , Adenocarcinoma , Fator de Transcrição CDX2 , Antígeno Ki-67RESUMO
ABSTRACT Background: Colorectal cancer (CRC) is one of the most common types of cancer in the world. Over time, intestinal epithelial cells undergo mutations that may lead to proliferative advantage and the emergence of cancer. Mutations in the beta-catenin pathway are amongst those described in the development of CRC. Aim: To verify the existence of a relation between the presence of Wnt3, beta-catenin and CDX2 in colorectal cancer samples and clinical outcomes such as disease progression or death. Method: Wnt3a, beta-catenin and CDX2 immunohistochemistry was performed on CRC tissue microarray samples (n=122), and analysis regarding the relation between biomarker expression and disease progression or death was performed. Results: No significant difference was found between the presence or absence of CDX2, beta-catenin or Wnt3a expression and clinical stage, tumor grade, disease progression or death. Conclusion: CDX2, beta-catenin and Wnt3a are not useful to predict prognosis in patients with CRC.
RESUMO Racional: O câncer colorretal (CCR) é um dos tipos mais comuns no mundo. As células epiteliais intestinais podem sofrer mutações que ocasionam vantagem proliferativa e culminam com o surgimento do câncer. Mutações da via da beta-catenina foram descritas entre as que podem ocasioná-lo. Objetivo: Verificar a existência de relação entre a expressão de Wnt3, beta-catenina e CDX2 em amostras de câncer colorretal com os eventos clínicos progressão de doença e óbito. Método: Foi realizada análise imunoistoquímica de Wnt3a, beta-catenina e CDX2 em blocos multiamostrais de CRC (n=122), e avaliada a relação entre a expressão dos biomarcadores e os desfechos progressão de doença e óbito. Resultados: Não foram encontradas diferenças significativas entre a expressão ou ausência de CDX2, beta-catenina ou Wnt3a e estádio clínico, grau de diferenciação tumoral, presença de progressão de doença ou evolução ao óbito. Conclusão: Os marcadores CDX2, beta-catenina e Wnt3a não são úteis para predizer prognóstico em pacientes com CCR.
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Humanos , Neoplasias Colorretais/diagnóstico , beta Catenina/genética , Proteína Wnt3/genética , Fator de Transcrição CDX2/genética , Imuno-Histoquímica , Neoplasias Colorretais/genética , Progressão da DoençaRESUMO
AIM: The aim of the present study was to evaluate the distribution of single-nucleotide polymorphisms (SNP) (variants FOKI [rs2228570], CDX2 [rs47908762], and GATA [rs4516035]) in the vitamin D receptor in individuals with type 2 diabetes mellitus and chronic periodontitis (DM2 + CP), CP alone, and healthy individuals, and to investigate the relationship with susceptibility to CP. METHODS: In total, 280 individuals (116 with DM2 + CP, 95 with CP alone, and 69 healthy individuals) were genotyped using real-time polymerase chain reaction with allele-specific probes. Significant differences (P < .05) were found among the groups with regard to socio-epidemiological variables (sex, marital status, income, smoking habit, and schooling) and clinical-epidemiological variables (age, number of teeth, probing depth, clinical attachment loss, gingival bleeding index, and visible plaque index). RESULTS: The C allele was significantly more frequent among the healthy individuals (34.8%) than those with DM2 + CP (23.5%) (odds ratio [OR] = .58, 95% confidence interval [CI]: . 35-.94, P = .022). Likewise, the CC allele was significantly more frequent among healthy individuals (11.6%) than those with DM2 + CP (2.6%) (OR = .17, 95% CI: .03-.79, P = .015). CONCLUSION: The results suggest that the presence of these variants could lead to a lower susceptibility to DM2 and CP. No other significant differences among groups were found for the other SNP investigated.
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Fator de Transcrição CDX2/genética , Periodontite Crônica/genética , Diabetes Mellitus Tipo 2/genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Receptores de Calcitriol/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Alelos , Brasil , Estudos de Casos e Controles , Periodontite Crônica/complicações , Periodontite Crônica/epidemiologia , Índice de Placa Dentária , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/epidemiologia , Feminino , Genótipo , Técnicas de Genotipagem , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Adulto JovemRESUMO
RESUMO Objetivo: Verificar a relação dos polimorfismos do gene do receptor de vitamina D (RVD) com sinais clínicos e níveis de vitamina D (VD) em asmáticos. Métodos: Estudo transversal com 77 crianças de 7 a 14 anos de um ambulatório especializado, divididas em 3 grupos: asmáticos, em uso de corticoide inalatório (ICS) por mais de um ano; asmáticos sem necessidade de ICS; não asmáticos e não alérgicos (de acordo com o International Study of Asthma and Allergies in Childhood - ISAAC. Foram avaliados: espirometria, testes alérgicos, presença do polimorfismo CDX2 do promotor do RVD por reação em cadeia da polimerase (PCR) e genotipagem de polimorfismos dos éxons 2 e 3 por PCR-SSCA (single-strand conformational analysis), imunoglobulina E (IgE) total e IgE específica para ácaros e gramíneas nos três grupos estudados. Níveis de 25-hidroxivitamina D foram dosados nos asmáticos. Resultados: A média de idade foi 10,8±2,2 anos, 57% masculinos, 38 asmáticos com ICS, 22 sem ICS e 17 não asmáticos. Rinite alérgica esteve presente em 90% dos asmáticos, polimorfismo CDX2 em 23% dos asmáticos e ausente nos controles (p=0,03). Menores níveis de volume expiratório forçado no primeiro segundo (VEF1%) foram observados nos asmáticos homozigotos para CDX2 (p=0,001). Variações nas sequências dos éxons 2 e 3 não foram relacionadas com a asma ou demais testes. Deficiência ou insuficiência de VD foi diagnosticada em 98% dos asmáticos. Não houve associação entre níveis de VD e polimorfismos genéticos dos éxons 2 e 3. Conclusões: Observou-se associação positiva entre polimorfismo CDX2 em homozigoze com asma e menores valores de VEF1%. O CDX2 pode modificar a interação celular do RVD com a vitamina, bem como pode estar associado com a asma e com a dificuldade de controle da doença.
ABSTRACT Objective: To verify the relationship between polymorphisms of the vitamin D receptor gene (VDR), clinical findings, and serum vitamin D (VD) levels in asthmatics. Methods: A cross sectional study of 77 children aged 7 to 14 years old, who were attended at a specialized clinic. The children were divided into 3 groups: asthmatics who had been using inhaled corticosteroids (ICS) for more than one year; asthmatics who had not been using ICS; non-asthmatics, and children without allergies (according to the International Study of Asthma and Allergies in Childhood - ISAAC). Spirometry, skin prick tests, the presence of a VDR promoter CDX2 polymorphism from an allele-specific polimerase chain reaction (PCR), exons 2 and 3 polymorphisms genotyping by PCR-SSCA (single-strand conformational analysis), total immunoglobulin E (IgE) and specific IgE to mites and grass were evaluated in these three groups. Levels of 25-hydroxyvitamin D were determined in asthmatics only. Results: The mean age of the children was 10.8±2.0 years old, 57% were male, 38 were asthmatic and using ICS, 22 were asthmatic and not using ICS, and 17 were non-asthmatic. Allergic rhinitis was present in 90% of asthmatics. Homozygous CDX2 was detected in 23% of the patients and absent in the control group (p=0.03). Lower forced expiratory volume in 1 second (FEV1%) values were observed in CDX2 homozygous asthmatics (p=0.001). Variations in the exon 2 and 3 sequences were not related to asthma or the other tests. VD deficiency or insufficiency was detected in 98% of asthmatics. There was no association between VD levels and genetic polymorphisms from exons 2 and 3. Conclusions: There was a positive association between homozygous CDX2 polymorphism, asthma and lower FEV1% values. CDX2 is capable of modifying cell interaction between VDR and VD, and it could be associated with the prevalence of asthma, and the difficulty in controlling the disease.
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Humanos , Masculino , Feminino , Criança , Adolescente , Asma/sangue , Receptores de Calcitriol/genética , Polimorfismo Genético , Asma/tratamento farmacológico , Vitamina D/sangue , Cálcio/sangue , Estudos Transversais , Corticosteroides/uso terapêutico , MutaçãoRESUMO
RESUMEN De acuerdo a la historia natural del cáncer del cuello uterino, en donde las lesiones preneoplásicas de bajo y alto grado pueden presentar fenómenos de regresión o progresión, existe gran interés en la búsqueda de biomarcadores que permita predecir la evolución de las lesiones preneoplásicas del cérvix hacia la progresión o regresión de la enfermedad. Estos biomarcadores pudieran ser de origen genético, o epigenético que alteren la expresión de los genes y que pudieran estar asociados con la carcinogénesis en diferentes tipos de tejido humano. El objetivo del estudio fue analizar la expresión del mARN de los genes SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 y HAND1 en muestras negativas para lesiones intraepiteliales cervicales (n=9), muestras con lesiones intraepiteliales de bajo grado (n=10) y alto grado (n=11). Se realizó análisis de expresión de los genes mencionados mediante qRT-PCR y el análisis de los datos se realizó mediante la prueba no paramétrica de ANOVA. La diferencia estadística se determinó en valores p< 0,05. Para los genes EDNRB y CDX2 se observó disminución 66,7% en las muestras sin alteraciones histológicas cervicales, comparado con una disminución en la expresión del 50% en muestras con LIEBG y para el grupo de LIEAG del 36,4% para el gen EDNRB y del 27,3% para el gen CDX2 dando una diferencia estadísticamente significativa p= 0,02. Sugiriendo que EDNRB y CDX2 podrían ser útiles como posibles biomarcadores en la carcinogénesis cervical.
ABSTRACT According into account the natural history of cervical cancer, where low- and high-grade preneoplastic lesions may present regression or progression phenomena, there is great interest in the search for biomarkers to predict the behavior of preneoplastic lesions of the cervix. These biomarkers may be of genetic origin, or epigenetics that alter the expression of genes and that may be associated with carcinogenesis in different types of human tissue. The objective of the study was to analyze the expression of the mRNA of the SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 and HAND1 genes in samples negative for cervical intraepithelial lesions (n = 9), low grade intraepithelial lesions (n=10) and high grade (n = 11). Expression analysis of the mentioned genes was performed using qRT-PCR and data analysis was performed using the non-parametric ANOVA test. The statistical difference was determined in values p <0.05. For the EDNRB and CDX2 genes, a 66.7% decrease was observed in the samples without cervical histological alterations, compared to a decrease in expression of 50% in LIEBG samples and 36.4% in the LIEAG group for the EDNRB gene And 27.3% for the CDX2 gene giving a statistically significant difference p = 0.02. Suggesting that EDNRB and CDX2 could be useful as potential biomarkers in cervical carcinogenesis.
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Preterm birth (PTB) is featured by less than 37weeks of gestational age or fewer than 259days since the first day from the last menstrual period. Complications of PTB are the major cause of neonatal deaths, several factors are linked to PTB increased risk including immunological and genetics. Vitamin D plays an important role in immune response modulation and its action occurs through the vitamin D receptor (VDR), which recently has been described as overexpressed in human placenta during the pregnancy. Herein we assessed two single nucleotide polymorphisms (SNPs) FokI (rs2228570 A>G) and Cdx-2 (rs11568820 T>C), within VDR, using TaqMan fluorogenic probes, and differential susceptibility to SPTB. We assessed 104 pregnant women with SPTB and 85 women with normal birth in a Northeastern Brazilian population. Statistically significant differences for both SNPs where found when comparing allele and genotype frequencies in both groups: the T allele for rs2228570 and A allele for rs11568820 were significantly more frequent in SPTB group than in normal birth group (p=0.000013 and p=0.00466, respectively). The rs11568820 A/A genotype was associated to clinical/demographic variables such as: premature birth (p=0.007), neonate weight (p=0.039), presence of infection during pregnancy (p=0.011) and premature birth among multiparous (p=0.015). The rs2228570 T/T genotype associated with gestational diabetes mellitus (p=0.044) and chorioamnionitis during pregnancy (p=0.043). In conclusion our findings indicate an association between polymorphisms FokI and Cdx-2 within VDR gene and SPTB, suggesting their involvement in the triggering of these syndromes.
Assuntos
Polimorfismo de Nucleotídeo Único , Nascimento Prematuro/genética , Receptores de Calcitriol/genética , Brasil , Estudos de Casos e Controles , Feminino , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Gravidez , Fatores de RiscoRESUMO
Acase of intestinal mucinous adenocarcinoma with metastasis to gonadal tissue is reported. A13-year-old, male, poodle dog presented with intestinal and peritoneal masses, as well as infiltrative masses in testicular tunics. Samples werebiopsied and submitted for histopathological analysis. Microscopically, intestinal lesion consisted of an adenocarcinoma (mucinous type), with infiltration of muscular layers and mesenteric adipose tissue. In gonadal tissue, there was neoplastic infiltration of epididymis and tunica albuginea (with a predominantly tubular pattern), and testicular parenchyma (with a predominantly signet-ring cell pattern). Immunohistochemistry was positive for CDX2 and pancytokeratin, and negative for vimentin, supporting the diagnosis of intestinal mucinous adenocarcinoma with metastases to epididymis, testis and tunica albuginea.(AU)
Assuntos
Animais , Masculino , Cães , Adenocarcinoma Mucinoso/veterinária , Metástase Neoplásica , Epididimo/patologia , Neoplasias Testiculares/veterinária , Imuno-Histoquímica/veterinária , Fator de Transcrição CDX2RESUMO
Acase of intestinal mucinous adenocarcinoma with metastasis to gonadal tissue is reported. A13-year-old, male, poodle dog presented with intestinal and peritoneal masses, as well as infiltrative masses in testicular tunics. Samples werebiopsied and submitted for histopathological analysis. Microscopically, intestinal lesion consisted of an adenocarcinoma (mucinous type), with infiltration of muscular layers and mesenteric adipose tissue. In gonadal tissue, there was neoplastic infiltration of epididymis and tunica albuginea (with a predominantly tubular pattern), and testicular parenchyma (with a predominantly signet-ring cell pattern). Immunohistochemistry was positive for CDX2 and pancytokeratin, and negative for vimentin, supporting the diagnosis of intestinal mucinous adenocarcinoma with metastases to epididymis, testis and tunica albuginea.
Assuntos
Masculino , Animais , Cães , Adenocarcinoma Mucinoso/veterinária , Epididimo/patologia , Metástase Neoplásica , Neoplasias Testiculares/veterinária , Imuno-Histoquímica/veterináriaRESUMO
Reduced CDX2 and cytokeratin 20 (CK20) expression in colorectal carcinoma with BRAF mutation and high-level microsatellite instability (MSI-H) has been well documented. The immunophenotype of BRAF-mutated microsatellite stable (MSS) colorectal carcinoma has not been reported. We analyzed 205 colorectal carcinomas including 28 BRAF-mutated MSS, 53 BRAF-mutated MSI-H, and 124 BRAF wild-type MSS tumors for CDX2, cytokeratin 7 (CK7), and CK20 immunohistochemical expression. CDX2 was scored semiquantitatively for both staining intensity and percent of tumor cells staining and a modified CDX2 H-score was calculated. Patients with BRAF-mutated MSS colorectal carcinomas were more frequently stage IV at presentation compared to patients with BRAF-mutated MSI-H colorectal carcinomas and BRAF wild-type MSS colorectal carcinomas (32% versus 8% versus 15%, P < .001). BRAF-mutated MSS colorectal carcinoma displayed reduced CDX2 expression compared to BRAF wild-type MSS colorectal carcinoma (75% versus 94%; mean CDX2 H-score 98 versus 150, P < .001). CK7 expression was more often identified in BRAF-mutated MSS colorectal carcinoma compared to both BRAF-mutated MSI-H colorectal carcinoma and BRAF wild-type MSS colorectal carcinoma (39% versus 6% versus 6%, P = .0001). BRAF-mutated MSI-H colorectal carcinomas were less often CK20 positive compared to BRAF-mutated MSS and BRAF wild-type MSS tumors (70% versus 93% versus 90%, P = 0.001). In summary, BRAF-mutated MSS colorectal carcinoma often displays reduced CDX2 and increased CK7 expression. Knowledge of this altered immunophenotype is important as patients with BRAF-mutated MSS colorectal carcinoma often present with metastatic disease and the altered tumor immunophenotype may lead to the erroneous assumption that origin from the colon/rectum is unlikely.
Assuntos
Adenocarcinoma/metabolismo , Neoplasias Colorretais/metabolismo , Proteínas de Homeodomínio/metabolismo , Queratina-7/metabolismo , Proteínas Proto-Oncogênicas B-raf/metabolismo , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Fator de Transcrição CDX2 , Neoplasias Colorretais/patologia , Feminino , Humanos , Imuno-Histoquímica , Queratina-20/metabolismo , Masculino , Instabilidade de Microssatélites , Repetições de Microssatélites , Pessoa de Meia-Idade , Mutação , PrognósticoRESUMO
INTRODUÇÃO: O esôfago de Barrett (EB) corresponde à substituição do epitélio escamoso por um do tipo intestinal, em resposta ao refluxo crônico nos pacientes com doença do refluxo gastroesofágico (DRGE). É um importante precursor do adenocarcinoma esofágico. A fundoplicatura de Nissen (FN) é uma cirurgia antirrefluxo que visa a reduzir a agressão à mucosa esofágica. Alterações no padrão de expressão imuno-histoquímica de mucinas e de CDX2 no EB antes e depois da FN podem ser úteis na identificação de um padrão de expressão desses marcadores e, eventualmente, na identificação de casos com risco de evolução para malignidade. OBJETIVOS: Avaliar e comparar a imunoexpressão de CDX2 e mucinas no EB de pacientes com DRGE submetidos à FN antes e após a cirurgia. MATERIAIS E MÉTODOS: Estudo retrospectivo de 25 pacientes com diagnóstico de DRGE e EB submetidos à FN, acompanhados por, pelo menos, três anos. Foram feitos análise histológica e estudo imuno-histoquímico das biópsias endoscópicas antes e após a cirurgia, comparando-se a inflamação e a imunoexpressão de MUC1, MUC2, MUC5AC, MUC6 e CDX2. Estimou-se a porcentagem de células com expressão para os marcadores estudados na mucosa de Barrett: 0%-25%, 25%-75% e 75%-100% das células positivas. Foram utilizados os testes de McNemar e Stuart-William e adotou-se o nível de 5% de significância estatística. RESULTADOS E CONCLUSÃO: Não houve diferenças significativas quanto a presença ou intensidade de inflamação, nem da imunoexpressão de mucinas e CDX2 no EB antes e após a FN. O tratamento cirúrgico não influenciou a mudança da expressão dessas glicoproteínas no EB.
INTRODUCTION: Barrett´s esophagus (BE) is characterized by the exchange of esophageal squamous epithelium for intestinal type in response to chronic reflux in patients with gastroesophageal reflux disease (GERD).It is an important precursor of esophageal adenocarcinoma. Nissen fundoplication (NF) is an antireflux surgery which aims to reduce esophageal mucosa inflammation. Changes in the immunohistochemical expression patterns of mucins (MUC1, MUC2, MUC5AC and MUC6) and CDX2 in BE before and after NF may be useful to identify the expression patterns of these markers and, possibly, to detect cases with risks of malignancy. OBJECTIVES: To investigate and compare mucin and CDX2 immunoexpression in BE patients with GERD before and after NF. MATERIAL AND METHODS: This retrospective study comprised 25 patients with GERD and BE who had been submitted to NF. The patients had a 3-year minimum follow up. Histological and immunohistochemical analyses of endoscopic biopsies were performed before and after the surgery, comparing inflammation and MUC1, MUC2, MUC5AC, MUC6 and CDX2 immunoexpression. The percentage of Barrett mucosa cells with expression to the studied markers was estimated at 0%-25%, 25%-75% and 75%-100%. McNemar and Stuart-William tests were used and the significance level of <0.05 was applied. RESULTS AND CONCLUSION: Concerning the presence or the intensity of inflammation and mucin and CDX2 expression in BE, there were no significant differences before and after NF. The surgical procedure did not promote any changes in the expression of these glycoproteins in BE.
Assuntos
Humanos , Esôfago de Barrett/genética , Fundoplicatura , Imuno-Histoquímica , Mucinas/genética , Refluxo Gastroesofágico/cirurgia , Refluxo Gastroesofágico/genéticaRESUMO
The micropapillary carcinoma is regarded as an aggressive variant of adenocarcinoma in any location. Histologically is characterized by papillary cell clusters surrounded by clear spaces. The reported proportion of micropapillary carcinoma component to the entire tumor ranged from 5 to 80 percent and no pure cases has been reported. There are near of 130 cases reported to date in colorectum. We experienced a patient with a pure micropapillary carcinoma showing coexpression of CK7, CK20, and absence of CDX2, which had an aggressive neoplasm with extense perineural, vascular and lymphatic invasion also extensive nodal metastasis. The presence of a micropapillary carcinoma in the colorectum seemed to be closely related with nodal metastasis, similar to the case for micropapillary carcinomas in other organs. Therefore, if a micropapillary component is identified in a tumor, particularly in a biopsy specimen, even if the pre-operative diagnosis is a pedunculated early colorectal cancer, should be carefully consider the extent of surgical resection due to the high potential for nodal metastasis.
El carcinoma micropapilar es considerado como una variante agresiva del cáncer en cualquier localización. Histológicamente se caracteriza por grupos de células papilares rodeada de espacios libres. Se informó que la proporción del componente carcinoma micropapilar en la totalidad de un tumor varió entre 5 por ciento a 80 por ciento y no se han reportado casos puros. Existen cerca de 130 casos reportados hasta la fecha en colon y recto. Se describe el caso de un paciente con un carcinoma micropapilar puro que muestra coexpresión de CK7, CK20, y la ausencia de CDX2, que tenía un tumor agresivo con extensa invasión perineural, vascular y linfática además de metástasis nodular extensa. La presencia de un carcinoma micropapilar en la región colorrectal parece estar estrechamente relacionada con metástasis nodular, similar al caso del carcinomas micropapilar en otros órganos. Por lo tanto, si un componente micropapilar se identifica en un tumor, sobre todo en una muestra de biopsia, incluso si el diagnóstico pre-operatorio es un cáncer colorrectal temprano pediculado, se debe considerar cuidadosamente la extensión de la resección quirúrgica debido a la alta probabilidad de metástasis nodular.
Assuntos
Idoso , Carcinoma Papilar , Neoplasias Retais/enzimologia , Neoplasias Retais/ultraestrutura , Adenocarcinoma/enzimologia , Adenocarcinoma/ultraestrutura , Regulação Neoplásica da Expressão GênicaRESUMO
Neste relato, aproveitamos um caso pouco usual de adenocarcinoma em mucosa sinonasal para ilustrar o processo de decisão diagnóstica em imuno-histoquímica. Trata-se de paciente do sexo masculino, de 86 anos, apresentando massa intranasal. No estudo imuno-histoquímico, notou-se expressão de antígeno prostático específico (PSA) e do CDX-2, indicando possibilidades diagnósticas divergentes, de metástase de adenocarcinoma prostático ou de adenocarcinoma sinonasal do tipo intestinal (ITAC). A partir desse dilema diagnóstico, buscou-se estabelecer como esses marcadores poderiam ser confrontados em uma decisão diagnóstica, definindo-se as qualidades que um bom marcador imuno-histoquímico deve ter, e como os marcadores utilizados no painel se comparam em relação a essas qualidades e às hipóteses diagnósticas levantadas. Nesse exercício, são desvendados aspectos importantes do processo diagnóstico em patologia.
In this report, we explore an unusual case of adenocarcinoma detected in the sinonasal mucous membrane in order to illustrate the diagnostic decision process in the immunohistochemical diagnosis. The patient is a 86-year-old man with an intranasal mass. In the immunohistochemical study, it was found expression for both PSA and CDX-2, raising the divergent diagnostic possibilities of a prostatic adenocarcinoma metastasis or an intestinal type sinonasal adenocarcinoma. In search for a solution for this diagnostic dilemma, we tried to establish how these immunohistochemical markers could be confronted in a diagnostic decision by defining the qualities that good immunohistochemical markers consist of and how the panel markers may be compared as to these qualities and the diagnostic hypotheses. In this exercise, important aspects of the diagnostic process in pathology are revealed.