Inmunoexpresión de E-cadherina y vimentina en mucosa oral normal, displasia epitelial oral y carcinoma oral de células escamosas / Immunoexpression of E-cadherin and vimentin in normal oral mucosa, oral epithelial displasia and oral squamous cell carcinoma

El objetivo fue evaluar la inmunoexpresión de E-cadherina y Vimentina en mucosa oral normal (MON), displasia epitelial oral (DEO) y carcinoma oral de células escamosas (COCE). Se realizó un estudio descriptivo de una serie de casos analizandolos mediante técnica de inmunohistoquímica contra E-cadherina y Vimentina 16 muestras de MON, 16 de DEO y 19 de COCE. La inmunotinción fue evaluada cualitativamente considerando extensión e intensidad para E-cadherina e intensidad para Vimentina. El análisis de la extensión e intensidad de la inmunotinción de E-cadherina y Vimentina según diagnóstico reveló una asociación estadísticamente significativa (p<0,001). Siendo la expresión de E-cadherina más alta en MON, seguido por DEO y más baja en COCE, inversamente a lo que se observó con Vimentina. El presente estudio reveló la subregulación del marcador molecular E-cadherina junto con la expresión aberrante por parte de células epiteliales del marcador mesenquimal Vimentina en muestras de MON, DEO y COCE.

The aim was to evaluate the expression of E-cadherin and Vimentin in oral epithelial dysplasia (OED) and oral squamous cell carcinoma (OSCC), in comparison with normal oral mucosa (NOM) in a descriptive case study using immunohistochemistry. A total of fifty-one (N=51) histological samples were included; as follows: n = 16 (NOM), n = 16 (OED) and n = 19 (OSCC). All samples were analyzed using immunohistochemistry against the expression of E-cadherin and Vimentin. Immunostaining was qualitatively evaluated by extent and intensity of its expression for E-cadherin and intensity for Vimentin. Extension and intensity analysis of E-cadherin and Vimentin immunostaining according to group revealed a statistically significant association (r<0.001). E-cadherin expression was found to be highest in NOM followed by OED and lowest in OSCC, inverse to what was observed with Vimentin. The present study revealed the down regulation of the molecular marker E-cadherin, suggestive of reduction in dysplastic cells on comparison to NOM cells, and aberrant expression of the mesenchymal marker Vimentin by epithelial cells in samples of NOM, OED and OSCC; questioning their value as a prognostic marker.

Efecto de las alteraciones genéticas y epigenéticas de la cadherina E y su expresión en la transcripción en la propensión al cáncer de mama / Effect of genetics, epigenetics and variations in the transcriptional expression of cadherin-E in breast cancer susceptibility

RESUMEN Introducción. La cadherina E (CDH1) cumple un papel importante en la transición epitelio-mesénquima y está relacionada con la invasión y las metástasis en varios tipos de carcinomas. Sin embargo, el efecto de las mutaciones y 'epimutaciones' germinales en la propensión al cáncer de mama no es claro. Objetivo. Evaluar el polimorfismo rs5030625, los cambios en el patrón de metilación del promotor y la expresión en la transcripción del gen CDH1 en pacientes con cáncer de mama. Materiales y métodos. Se tomaron muestras de sangre periférica de 102 pacientes con cáncer de mama y 102 mujeres de control. La genotipificación del polimorfismo rs5030625 se hizo mediante reacción en cadena de la polimerasa (PCR) y análisis de polimorfismos de longitud del fragmento de restricción; la PCR y el análisis de disociación de alta resolución sensible a metilación se emplearon para determinar el estado y el nivel de metilación del promotor del CDH1; por último, el nivel de expresión en la transcripción del CDH1 se evaluó mediante PCR cuantitativa con transcripción inversa. Resultados. Los resultados no evidenciaron asociación entre el polimorfismo rs5030625 y el cáncer de mama. Se encontraron perfiles aberrantes de metilación del promotor del CDH1 en las pacientes con cáncer de mama relacionados con las primeras etapas de desarrollo del cáncer. La disminución de la expresión del CDH1 se asoció con la presencia de metástasis y el estado de metilación del promotor. Conclusión. Las alteraciones en el CDH1 se asociaron con la invasión y las metástasis en el cáncer de mama. Se proporcionó evidencia adicional sobre la relevancia del CDH1 en el desarrollo y la progresión del cáncer de mama.

ABSTRACT Introduction: Cadherin-E (CDH1) is an important regulator of epithelial-mesenchymal transition, invasion and metastasis in many carcinomas. However, germinal epimutations and mutations effect in breast cancer susceptibility is not clear. Objective: To evaluate rs334558 polymorphism, promoter methylation status and CDH1 expression profile in breast cancer patients. Materials and methods: We collected peripheral blood samples from 102 breast cancer patients and 102 healthy subjects. The identification of rs334558 polymorphism was performed using PCR-RFLP, while methylation-specific PCR (MSP) and methylation-sensitive high-resolution melting (MS-HRM) were used to explore CDH1 methylation status; finally, CDH1 transcriptional expression profile was evaluated using RT-qPCR. Results: We found no association between rs334558 polymorphism and breast cancer. Aberrant promoter methylation profile was found in breast cancer patients and it was related with early cancer stages. CDH1 down-regulation was significantly associated with metastasis and promoter methylation. Conclusion: CDH1 alterations were associated with invasion and metastasis in breast cancer. Our results offer further evidence of CDH1 relevance in breast cancer development and progression.

Epithelial-mesenchymal transition (EMT): principles and clinical impact in cancer therapy / Transición epitelio-mesenquimática (TEM): principios e impacto clínico en la terapia contra el cáncer

The epithelial-mesenchymal transition (EMT) is a biological phenomenon responsible for the formation of different tissues and organs during normal metazoan development. Because of the connection of the EMT with the pathogenesis of certain diseases, such as cancer, the attention of the scientific community has been directed towards the search for and identification of effective therapeutic targets. These targets include signal transduction in cancerous stem cells and the use of microRNAs, which would inhibit EMT-associated phenotypic changes and tumoral progression. In an attempt to compile relevant and current information, this work addresses concepts that define the EMT and the advances in this field. The wealth of knowledge gained from areas such as the loss of cell polarity and intracellular adhesion complexes, the signaling pathways implicated, microRNA participation in this process, and stemness acquisition in embryonic and cancerous cells, all of which allow for the visualization of promising perspectives, particularly, methods for targeting advanced malignancies, are presented herein.

La transición epitelio-mesenquimática (TEM) es el fenómeno biológico responsable de la formación de los diferentes tejidos y órganos durante el desarrollo normal de los organismos metazoarios. En razón de su conexión con la patogénesis de ciertas enfermedades como el cáncer, la atención de la comunidad científica se ha redireccionado hacia la búsqueda e identificación de blancos terapéuticos efectivos, como la transducción de señales de las células madre cancerosas o la utilización de microARNs, que permitirían bloquear los cambios fenotípicos asociados con la TEM y, por ende, la progresión tumoral. En un intento por recopilar información relevante y actualizada, el presente trabajo aborda conceptos que definen a la TEM y avances alcanzados en este campo. El acervo de conocimiento obtenido en aspectos como pérdida de la polaridad celular y de los complejos de adhesión intercelular, vías de señalización implicadas y participación de los microARNs en el proceso, así como adquisición de stemness o troncalidad, tanto en células embrionarias como cancerosas, hace posible visualizar perspectivas promisorias, en especial en lo que se refiere a las terapias contra las malignidades de alto grado.

Las cadherinas en el diagnostico histopatológico y pronostico del carcinoma de células renales / The cadherins in the histopathology diagnosis and prognosis of renal cell carcinoma

El carcinoma de células renales es el cáncer renal más común en el adulto. Se describen 5 tipos histológicos principales, que, en general, muestran al microscopio óptico características que permiten diagnosticarlos. Sin embargo, el solapamiento morfológico observado entre los diferentes tipos y la heterogeneidad histológica dentro del mismo tumor, continúan generando problemas en su clasificación. Algunos marcadores inmunohistoquímicos son empleados en el diagnóstico diferencial de estos carcinomas, pero tienen sus limitaciones. En el presente trabajo, se hizo una revisión de varios estudios, en los que se identificaron alteraciones en la expresión de algunas cadherinas en tejidos renales y líneas celulares con carcinoma de células renales. Estas alteraciones estuvieron asociadas con diferentes estados del desarrollo y progresión del tumor, y sirvieron para predecir el comportamiento neoplásico. Incrementar nuestro conocimiento en este campo, contribuiría al hallazgo de marcadores mucho más específicos para el diagnóstico y pronóstico de estos tumores.

Renal cell carcinoma is the most common adult renal cancer. Five main histological types are described, that in general display characteristic light microscopic features that lead to a correct diagnosis. However, the morphologic overlap between tumors and the histologic heterogeneity within a single tumor continue to create problems in the classification. Some immunohistochemical markers are used in the differential diagnosis of these carcinomas, but they have limitations. In the present work, a review of several studies was made, in which alterations in expression of some cadherins in renal tissue and cell lines with renal cell carcinoma were identified. These alterations were associated with different stages of tumor development and progression, and used to predict the neoplastic behaviour. To increase our knowledge in this field, will contribute to find more specific markers for the diagnosis and prognosis of these tumors.

Expression of cell adhesion molecules in the normal and T3 blocked development of the tadpole's kidney of Bufo arenarum (Amphibian, Anuran, Bufonidae) / Expressão das moléculas de adesão celular no desenvolvimento normal e com a inibição do hormônio tireoidea do rim nas larvas do Bufo arenarum (Amphibia, Anura, Bufonidae)

Cell adhesion molecules act as signal transducers from the extracellular environment to the cytoskeleton and the nucleus and consequently induce changes in the expression pattern of structural proteins. In this study, we showed the effect of thyroid hormone (TH) inhibition and arrest of metamorphosis on the expression of E-cadherin, β-and α-catenin in the developing kidney of Bufo arenarum. Cell adhesion molecules have selective temporal and spatial expression during development suggesting a specific role in nephrogenesis. In order to study mechanisms controlling the expression of adhesion molecules during renal development, we blocked the B. arenarum metamorphosis with a goitrogenic substance that blocks TH synthesis. E-cadherin expression in the proximal tubules is independent of thyroid control. However, the blockage of TH synthesis causes up-regulation of E-cadherin in the collecting ducts, the distal tubules and the glomeruli. The expression of β-and α-catenin in the collecting ducts, the distal tubules, the glomeruli and the mesonephric mesenchyme is independent of TH. TH blockage causes up-regulation of β-and α-catenin in the proximal tubules. In contrast to E-cadherin, the expression of the desmosomal cadherin desmoglein 1 (Dsg-1) is absent in the control of the larvae kidney during metamorphosis and is expressed in some interstitial cells in the KClO4 treated larvae. According to this work, the Dsg-1 expression is down-regulated by TH. We demonstrated that the expression of E-cadherin, Dsg-1, β-catenin and α-catenin are differentially affected by TH levels, suggesting a hormone-dependent role of these proteins in the B. arenarum renal metamorphosis.

Moléculas de adesão celular atuam como tradutores do ambiente extracelular para o citoesqueleto e o núcleo e, conseqüentemente, induzindo mudanças no padrão da expressão das proteínas estruturais. Neste estudo, observamos os efeitos da inibição do hormônio tireóidea (TH) e detenção da metamorfose na expressão da E-caderina, β- e α- catenina no desenvolvimento do rim do Bufo arenarum. As moléculas de adesão celular durante o desenvolvimento têm uma expressão temporal e espacial seletiva, sugerindo um papel específico na nefrogênese. Com o propósito de estudar os mecanismos de controle da expressão das moléculas de adesão durante o desenvolvimento renal, bloqueou-se a metamorfose do B. arenarum com uma substancia goitrogênica que bloqueia a síntese de TH. A expressão da E-caderina nos tubos proximais é independente do controle da tireóide. Entretanto, o bloqueio da síntese de TH provoca uma sobre elevação da E-caderina nos dutos coletores, nos tubos distais e nos glomérulos. A expressão da β- e α-catenina nos dutos coletores, nos tubos distais, nos glomérulos e no mesênquima mesonéfrico é independente da TH. O bloqueio da TH causa uma sobre-regulação da β- e α-catenina nos tubos proximais. Em contraste com a E-caderina, a expressão da caderina desmossomal demogloína 1 (Dsg-1) é ausente no controle durante a metamorfose da fase larval dos rins e se expressa em algumas células intersticiais nas larvas tratadas com KClO4. De acordo com este trabalho, a expressão Dsg-1 é subregulada pela TH. Demonstramos que a expressão da E-caderina, Dsg-1, β-catenina e α-catenina são afetadas de forma diferencial pelos níveis de TH, sugerindo um dependência hormonal destas proteínas na metamorfose renal do B. arenarum.

Expression of cell adhesion molecules in the normal and T3 blocked development of the tadpole's kidney of Bufo arenarum (Amphibian, Anuran, Bufonidae)

Cell adhesion molecules act as signal transducers from the extracellular environment to the cytoskeleton and the nucleus and consequently induce changes in the expression pattern of structural proteins. In this study, we showed the effect of thyroid hormone (TH) inhibition and arrest of metamorphosis on the expression of E-cadherin, -and -catenin in the developing kidney of Bufo arenarum. Cell adhesion molecules have selective temporal and spatial expression during development suggesting a specific role in nephrogenesis. In order to study mechanisms controlling the expression of adhesion molecules during renal development, we blocked the B. arenarum metamorphosis with a goitrogenic substance that blocks TH synthesis. E-cadherin expression in the proximal tubules is independent of thyroid control. However, the blockage of TH synthesis causes up-regulation of E-cadherin in the collecting ducts, the distal tubules and the glomeruli. The expression of -and -catenin in the collecting ducts, the distal tubules, the glomeruli and the mesonephric mesenchyme is independent of TH. TH blockage causes up-regulation of -and -catenin in the proximal tubules. In contrast to E-cadherin, the expression of the desmosomal cadherin desmoglein 1 (Dsg-1) is absent in the control of the larvae kidney during metamorphosis and is expressed in some interstitial cells in the KClO4 treated larvae. According to this work, the Dsg-1 expression is down-regulated by TH. We demonstrated that the expression of E-cadherin, Dsg-1, -catenin and -catenin are differentially affected by TH levels, suggesting a hormone-dependent role of these proteins in the B. arenarum renal metamorphosis.

Moléculas de adesão celular atuam como tradutores do ambiente extracelular para o citoesqueleto e o núcleo e, conseqüentemente, induzindo mudanças no padrão da expressão das proteínas estruturais. Neste estudo, observamos os efeitos da inibição do hormônio tireóidea (TH) e detenção da metamorfose na expressão da E-caderina, - e - catenina no desenvolvimento do rim do Bufo arenarum. As moléculas de adesão celular durante o desenvolvimento têm uma expressão temporal e espacial seletiva, sugerindo um papel específico na nefrogênese. Com o propósito de estudar os mecanismos de controle da expressão das moléculas de adesão durante o desenvolvimento renal, bloqueou-se a metamorfose do B. arenarum com uma substancia goitrogênica que bloqueia a síntese de TH. A expressão da E-caderina nos tubos proximais é independente do controle da tireóide. Entretanto, o bloqueio da síntese de TH provoca uma sobre elevação da E-caderina nos dutos coletores, nos tubos distais e nos glomérulos. A expressão da - e -catenina nos dutos coletores, nos tubos distais, nos glomérulos e no mesênquima mesonéfrico é independente da TH. O bloqueio da TH causa uma sobre-regulação da - e -catenina nos tubos proximais. Em contraste com a E-caderina, a expressão da caderina desmossomal demogloína 1 (Dsg-1) é ausente no controle durante a metamorfose da fase larval dos rins e se expressa em algumas células intersticiais nas larvas tratadas com KClO4. De acordo com este trabalho, a expressão Dsg-1 é subregulada pela TH. Demonstramos que a expressão da E-caderina, Dsg-1, -catenina e -catenina são afetadas de forma diferencial pelos níveis de TH, sugerindo um dependência hormonal destas proteínas na metamorfose renal do B. arenarum.