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SUMMARY: Cerebral ischemia has not only a high mortality rate, which is the second leading cause of death worldwide, but is also responsible for severe disabilities in working age individuals, generating enormous public expending for treatment and rehabilitation of the affected individuals. The role of microRNAs in the pathophysiology of cerebral ischemia has been highlighted in current investigations. In addition, recent studies have also highlighted physical exercise as a possible protective factor both in the prevention and in the effects of cerebral ischemia, placing it as an important study resource. Thus, we investigated the role of physical exercise in experimental cerebral ischemia associated with the expression of microRNA-27b. 16 animals were used, divided into four experimental groups: Control, Physical Exercise, Cerebral Ischemia and Cerebral Ischemia associated with Physical Exercise. The real-time PCR methodology was used to analyze the expression of microRNA-27b. Although there were no statistically significant differences in the expression of microRNA-27b between the groups studied, the increased expression of microRNA-27b in the Physical Exercise group indicates its neuroprotective role in the pathophysiology of cerebral ischemia.

RESUMEN: La isquemia cerebral no solo tiene una alta tasa de mortalidad y es la segunda causa principal de muerte en todo el mundo, sino también es la causa de enfermedades invalidantes en personas en edad laboral, lo que genera un gasto público enorme para el tratamiento y la rehabilitación de las personas afectadas. El papel de los microARN en la fisiopatología de la isquemia cerebral se ha destacado en las investigaciones actuales. Además, estudios recientes también han destacado el ejercicio físico como un posible factor protector tanto en la prevención como en los efectos de la isquemia cerebral, situándolo como un importante recurso de estudio. Por lo tanto, investigamos el papel del ejercicio físico en la isquemia cerebral experimental asociada con la expresión del microARN-27b. Se utilizaron 16 animales, divididos en cuatro grupos experimentales: Control, Ejercicio Físico, Isquemia Cerebral e Isquemia Cerebral asociada al Ejercicio Físico. Se utili- zó la metodología de PCR en tiempo real para analizar la expresión de microARN-27b. Aunque no se observaron diferencias estadísticamente significativas en la expresión de microARN-27b entre los grupos estudiados, la mayor expresión de microARN-27b en el grupo de Ejercicio Físico indica su papel neuroprotector en la fisiopatología de la isquemia cerebral.

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Expression of AIF, PARP, and miRNAs MIR-145, MIR-210, and MIR-486 Associated with Apoptosis in the Corpus Cavernosum Of Rats Subjected to Chronic Alcoholism Model
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Sarraipo, Vagner Schiavoni

; Lizarte Neto, Fermino Sanches

; Carvalho, Camila Albuquerque Mello de

; Silva, Jairo Pinheiro da

; Durand, Marina Toledo

; Schimming, Bruno Cezar

; Martinez, Marcelo

; Galli, Maria de Fátima Tazima Sorita

; Buono, Fabianna de Oliveira

; Tiezzi, Maria Fernanda Barbirato da Mata

; Novais, Paulo Cezar

; Gula, Isabella Stracieri

; Thomazini, José Antônio

; Fazan, Valeria Paula Sassoli

; Rodrigues, Andressa Romualdo

; Tirapelli, Daniela Pretti da Cunha

; Cirino, Mucio Luiz de Assis

; Molina, Carlos Augusto Fernandes

; Junior, Silvio Tucci

; Tirapelli, Luis Fernando

International Journal of Morphology

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dec 2020, Volume 38 N. 6 Pages 1639 - 1644

Abstract: En Es | Text: En Es | PDF: En | PDF: Es

SUMMARY: Previous studies from our group described the consequences of using ethanol on penile erection. Nevertheless, the molecular mechanisms surrounding microRNAs, apoptosis process and their relationship with erectile dysfunction associated with alcohol consumption are still poorly understood. The objective of this analysis was to evaluate the mechanism of apoptosis by the expression of AIF and PARP, as well as their regulatory microRNAs: miR-145, miR-210 and miR-486, in the corpus cavernosum of rats submitted to a semivoluntary alcoholism model. For this study 24 Wistar rats were divided into two groups: control (C) and treated with 20 % ethanol (A) for seven weeks. The corpus cavernosum samples were prepared for immunohistochemical analysis of AIF and PARP protein expression, and microRNAs miR-145, miR-210, miR-486 gene expression in cavernous tissue was performed by real time PCR. The immunohistochemical analysis showed little nuclear positive labeling for the protein PARP and AIF in the corpus cavernosum of control and ethanol treated animals. After analysis of miR-145, -210 and -486 microRNA expression in the 12 animals studied, no results were found with significant statistical difference between the control and alcoholized groups. The expression of AIF and PARP and their regulatory microRNAs involved in apoptotic process (miR-145, miR-210 and miR-486) were not altered in the corpus cavernosum of rats submitted to semivoluntary alcoholism.

RESUMEN: Estudios previos de nuestro grupo describieron las consecuencias del uso de etanol en la erección del pene. Sin embargo, los mecanismos moleculares que rodean a los microARN, el proceso de apoptosis y su relación con la disfunción eréctil asociada con el consumo de alcohol aún no se conocen bien. El objetivo de este análisis fue evaluar el mecanismo de apoptosis mediante la expresión de AIF y PARP, así como sus microARN reguladores: miR-145, miR-210 y miR-486, en el cuerpo cavernoso de ratas sometidas a un modelo de alcoholismo semivoluntario. Se dividieron 24 ratas Wistar en dos grupos: control (C) grupo de ratas tratadas con etanol al 20 % (A) durante siete semanas. Las muestras del cuerpo cavernoso se prepararon para el análisis inmunohistoquímico de la expresión de la proteína AIF y PARP, y la expresión del gen microRNAs miR-145, miR-210, miR-486 en tejido cavernoso se realizó por PCR en tiempo real. El análisis inmunohistoquímico mostró escaso etiquetado nuclear positivo para la proteína PARP y AIF en el cuerpo cavernoso de los animales de control y tratados con etanol. Después del análisis de la expresión de microARN miR-145, -210 y -486 no se encontraron resultados con diferencias estadísticas significativas entre los grupos control y alcoholizados. La expresión de AIF y PARP y sus microARN reguladores involucrados en el proceso apoptótico (miR-145, miR-210 y miR-486) no se alteraron en el cuerpo cavernoso de las ratas sometidas a alcoholismo semivoluntario.

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AIF, PARP-1 and MicroRNA-9 Expression in Cerebral Ischemia Associated to Alcoholism Experimental Model
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Abrahão, Dayana Pousa Siqueira

; Lizarte Neto, Fermino Sanches

; Rodrigues, Andressa Romualdo

; Cirino, Múcio Luiz de Assis

; Silva, Jairo Pinheiro da

; Sarraipo, Vagner Schiavoni

; Carvalho, Camila Albuquerque Melo de

; Tazima, Maria de Fátima Galli Sorita

; Carlotti-Jr., Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luís Fernando

; Tirapelli, Daniela Pretti da Cunha

International Journal of Morphology

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jun 2020, Volume 38 N. 3 Pages 616 - 621

Abstract: En Es | Text: En Es | PDF: En | PDF: Es

SUMMARY: The chronic consumption of alcohol causes a worsening of the events that follow the cerebral ischemia. These events are regulated through the expression of several genes and microRNAs. The aimof this work was To analyze and describe the expression profile of PARP and AIF and miRNA-9 proteins in rats submitted to focal cerebral ischemia, associated or not with chronic alcoholism model. Methods: Twenty adult Wistar rats, subdivided into: control; ischemic; alcoholic and ischemic / alcoholized for immunohistochemical analysis and miRNA-9 gene expression. Results: There was a reduction in the protein expression of PARP-1 and a positive marking for AIF in the ischemic / alcoholized group. The miRNA-9 did not obtain significant expression. The association of ischemia with chronic alcohol use promoted a tendency to low expression of miRNA-9, low expression of PARP-1 and high expression of AIF, indicating an interference in the protective effect of miRNA-9 be observed in the other groups.

RESUMEN: El consumo crónico de alcohol provoca un empeoramiento de los eventos que siguen a la isquemia cerebral. Estos eventos están regulados a través de la expresión de varios genes y microRNA. El objetivo de este trabajo fue analizar y describir el perfil de expresión de las proteínas PARP y AIF y microRNA-9 en ratas sometidas a isquemia cerebral focal, asociadas o no, con el modelo de alcoholismo crónico. Veinte ratas Wistar adultas se dividieron en: grupo control, isquémico alcohólico, e isquémico / alcoholizado para análisis inmunohistoquímico y expresión de genes microRNA-9. Resultados: Hubo una reducción en la expresión de proteínas de PARP-1 y un marcado positivo para AIF en el grupo isquémico / alcoholizado. No se observó una expresión significativa en el microRNA-9. La asociación de la isquemia con el consumo crónico de alcohol promovió una tendencia a la baja expresión de microRNA-9, baja expresión de PARP1 y alta expresión de AIF, lo que indica una interferencia en el efecto protector de microRNA-9 en los otros grupos.

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Chronic alcoholism associated with diabetes induced apoptosis in the corpus cavernosum of rats
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Schimming, Bruno Cesar

; Cirino, Mucio Luiz de Assis

; Lizarte Neto, Fermino Sanches

; Novais, Paulo Cezar

; Carvalho, Camila Albuquerque Melo de

; Tirapelli, Daniela Pretti da Cunha

; Molina, Carlos Augusto Fernandes

; Tirapelli, Luis Fernando

; Tucci Junior, Silvio

Acta Cirúrgica Brasileira

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2020, Volume 35 N. 8 elocation e202000805

Abstract: En | Text: En | PDF: En

Abstract Purpose To evaluate the effects of alcohol exposure and diabetes on apoptotic process in the corpus cavernosum. Methods Forty eight male Wistar rats were divided into four groups: control, diabetic, alcoholic and diabetic-alcoholic. Samples of the corpus cavernosum were prepared to study protein expression of apoptotic genes (Caspases-3 and 9) by immunohistochemistry and Real-Time PCR. Results The immunoreactivity of Caspases-3 and -9 was diffuse and higher in the treated groups though there was no significant difference between the experimental groups, only when compared with the control group. An increase was observed in the gene expression of Caspases-9 in the diabetic and ethanol-diabetic groups when compared with control and ethanol groups. Conclusions The association of these factors (ethanol and diabetes) probably can affect the apoptosis mechanism in lesions of the cavernous tissue in the rat penis. Both gene and protein expression of Caspase-9 in diabetic and ethanol-diabetic groups suggest the involvement of the apoptosis cascade from this study model.

https://doi.org/10.1590/s0102-865020200080000005 723 downloads

Morphological and molecular analysis of apoptosis in the corpus cavernosum of rats submitted to a chronic alcoholism model
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Meirelles, Rogério José de Azevedo

; Lizarte Neto, Fermino Sanches

; Cirino, Mucio Luiz de Assis

; Novais, Paulo Cezar

; Gula, Isabella Stracieri

; Silva, Jairo Pinheiro da

; Tazzima, Maria de Fátima Galli Sorita

; Fazan, Valéria Paula Sassoli

; Durand, Marina Toledo

; Tirapelli, Daniela Pretti da Cunha

; Carvalho, Camila Albuquerque Melo de

; Schimming, Bruno César

; Molina, Carlos Augusto Fernandes

; Tucci Junior, Silvio

; Tirapelli, Luis Fernando

Acta Cirúrgica Brasileira

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2020, Volume 35 N. 3 elocation e202000305

Abstract: En | Text: En | PDF: En

Abstract Purpose To evaluate the effect of chronic alcoholism on morphometry and apoptosis mechanism and correlate with miRNA-21 expression in the corpus cavernosum of rats. Methods Twenty-four rats were divided into two experimental groups: Control (C) and Alcoholic group (A). After two weeks of an adaptive phase, rats from group A received only ethanol solution (20%) during 7 weeks. The morphometric and caspase-3 immunohistochemistry analysis were performed in the corpus cavernosum. The miRNA-21 expression was analyzed in blood and cavernous tissue. Results Chronic ethanol consumption decreased cavernosal smooth muscle area of alcoholic rats. The protein expression of caspase 3 in the corpus cavernosum was higher in A compared to the C group. There was no difference in the expression of miRNA-21 in serum and cavernous tissue between the groups. Conclusion Chronic ethanol consumption reduced smooth muscle area and increased caspase 3 in the corpus cavernosum of rats, without altered serum and cavernosal miR-21 gene expression.

https://doi.org/10.1590/s0102-865020200030000005 715 downloads

PPARG expression in colorectal cancer and its association with staging and clinical evolution
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Villa, Andre Luiz Prezotto

; Parra, Rogério Serafim

; Feitosa, Marley Ribeiro

; Camargo, Hugo Parra de

; Machado, Vanessa Foresto

; Tirapelli, Daniela Pretti da Cunha

; Rocha, José Joaquim Ribeiro da

; Feres, Omar

Acta Cirúrgica Brasileira

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2020, Volume 35 N. 7 elocation e202000708

Abstract: En | Text: En | PDF: En

Abstract Purpose To evaluate the gene expression of peroxisome proliferator activated receptors gamma (PPARG) in colorectal tumors and to correlate this data with clinical variables of the patients. Methods We analyzed the gene expression of PPARG in 50 samples of colorectal tumors using real-time reverse transcription polymerase chain reaction, and 20 adjacent normal tissue samples as control. The results of these quantifications were correlated with the respective patients’ medical records’ clinical information. Results PPARG expression was not different in the tumor tissue compared to the control tissue. Patients older than 60 years, histological type with mucinous differentiation, more advanced staging at the time of diagnosis, and patients who evolved with recurrence of the disease or death did not present higher PPARG expression. Conclusion Expression of PPARGD was not associated with worse prognosis.

https://doi.org/10.1590/s0102-865020200070000008 724 downloads

Analysis of Caspase-9 protein and microRNAs miR-21, miR-126 and miR-155 related to the apoptosis mechanism in the cerebellum of rats submitted to focal cerebral ischemia associated with an alcoholism model
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SILVA, Jairo Pinheiro da

; LIZARTE NETO, Fermino Sanches

; CIRINO, Mucio Luiz de Assis

; CARVALHO, Camila Albuquerque Melo de

; CARLOTTI JR, Carlos Gilberto

; COLLI, Benedicto Oscar

; TIRAPELLI, Daniela Pretti da Cunha

; TIRAPELLI, Luís Fernando

Arquivos de Neuro-Psiquiatria

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oct 2019, Volume 77 N. 10 Pages 689 - 695

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO O objetivo deste estudo foi analisar o cerebelo de ratos submetidos à isquemia cerebral focal experimental, por oclusão da artéria cerebral média por 90 minutos, seguida de reperfusão por 48 horas, associada a um modelo de alcoolismo. Métodos Foram utilizados 50 ratos Wistar adultos, subdivididos em cinco grupos experimentais: grupo controle (C): animais submetidos apenas à anestesia; grupo sham (S): animais submetidos à simulação completa do procedimento cirúrgico; grupo isquêmico (I): animais submetidos à isquemia cerebral focal por 90 minutos, seguidos de reperfusão por 48 horas; grupo alcoólico (A): animais que receberam etanol absoluto diário diluído em 20% em água por quatro semanas; e grupo isquêmico e alcoólico (I + A): animais que recebem o mesmo tratamento do grupo A e, após quatro semanas, submetidos à isquemia cerebral focal por 90 minutos, seguidos de reperfusão por 48 horas. As amostras de cerebelo foram coletadas e a análise imuno-histoquímica da proteína Caspase-9 e a análise sérica por RT-PCR dos microRNAs miR-21, miR-126 e miR155 foram realizadas. Resultados A expressão de Caspase-9 foi maior nos grupos I, A e I + A. Nas análises de microRNAs, o miR-126 foi maior nos grupos A e I + A, o miR-155 foi maior nos grupos I e I + A. Conclusões Concluímos que a apoptose ocorre no córtex cerebelar, mesmo distante do foco isquêmico, e que os microRNAs 126 e 155 mostram uma correlação com a apoptose celular em ratos isquêmicos e submetidos ao modelo crônico de álcool.

ABSTRACT This study aimed to analyze the cerebellum of rats submitted to an experimental focal cerebral ischemia, by middle cerebral artery occlusion for 90 minutes, followed by reperfusion for 48 hours, associated with an alcoholism model. Methods Fifty adult Wistar rats were used, subdivided into five experimental groups: control group (C): animals submitted to anesthesia only; sham group (S): animals submitted to complete simulation of the surgical procedure; ischemic group (I): animals submitted to focal cerebral ischemia for 90 minutes followed by reperfusion for 48 hours; alcoholic group (A): animals that received daily absolute ethanol diluted 20% in water for four weeks; and, ischemic and alcoholic group (I + A): animals receiving the same treatment as group A and, after four weeks, submitted to focal cerebral ischemia for 90 minutes, followed by reperfusion for 48 hours. The cerebellum samples were collected and immunohistochemical analysis of Caspase-9 protein and serum analysis by RT-PCR of microRNAs miR-21, miR-126 and miR155 were performed. Results The expression of Caspase-9 was higher in groups I, A and I + A. In the microRNAs analyses, miR-126 was higher in groups A and I + A, miR-155 was higher in groups I and I + A. Conclusions We conclude that apoptosis occurs in the cerebellar cortex, even if it is distant from the ischemic focus, and that microRNAs 126 and 155 show a correlation with cellular apoptosis in ischemic rats and those submitted to the chronic alcohol model.

https://doi.org/10.1590/0004-282x20190126 1287 downloads

Morphological and immunohistochemical analysis of proteins CASPASE 3 and XIAP in rats subjected to cerebral ischemia and chronic alcoholism
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Schiavoni, Vagner Sarraipo

; Silva, Jairo Pinheiro da

; Lizarte Neto, Fermino Sanches

; Assis, Múcio Luiz Cirino de

; Tazima, Maria de Fátima Galli Sorita

; Carvalho, Camila Albuquerque Melo de

; Tirapelli, Daniela Pretti da Cunha

; Carlotti Jr, Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luis Fernando

Acta Cirúrgica Brasileira

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aug 2018, Volume 33 N. 8 Pages 652 - 663

Abstract: En | Text: En | PDF: En

Abstract Purpose: To evaluate histopathological and ultrastructural changes and expression of proteins related to apoptosis CASPASE 3 and XIAP after experimental induction of temporary focal cerebral ischemia (90 minutes) due to obstruction of the middle cerebral artery in alcoholism model. Methods: Forty adult Wistar rats were used, subdivided into 5 experimental groups: control group (C); Sham group (S); Ischemic group (I); Alcoholic group (A); and Ischemic and Alcoholized group (I+A): animals submitted to the same treatment of group A and after four weeks were submitted to focal cerebral ischemia during 90 minutes, followed by reperfusion of 48 hours. Were processed for histopathological analysis and immunohistochemistry (for the protein expression of CASPASE -3 and XIAP). Results: Greater histopathological changes were observed in the animals of groups I and I+A in the three areas analyzed. The neuronal loss was higher in the medial striatum region of the animals of groups I and I + A. The protein expression of CASPASE -3 was higher than that of XIAP in the groups I and I + A for both proteins. Conclusion: The expression of XIAP was slightly higher where the histopathological changes and expression of CASPASE -3 was less evident.

https://doi.org/10.1590/s0102-865020180080000001 877 downloads

High expression of XIAP and Bcl-2 may inhibit programmed cell death in glioblastomas
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Tirapelli, Daniela Pretti da Cunha

; Lustosa, Isis Lacrose

; Menezes, Sarah Bomfim

; Franco, Indira Maynart

; Rodrigues, Andressa Romualdo

; Peria, Fernanda Maris

; Marinho, Alexandre Magno da Nóbrega

; Serafini, Luciano Neder

; Carlotti Jr, Carlos Gilberto

; Tirapelli, Luís Fernando

Arquivos de Neuro-Psiquiatria

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dec 2017, Volume 75 N. 12 Pages 875 - 880

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO O glioblastoma (GBM) é o glioma mais maligno e representa 29% de todos os tumores cerebrais. A tumorigênese está intimamente ligada à características adquiridas na via fisiológica de morte celular. Objetivo: Avaliar a expressão de genes anti-apoptóticos (XIAP e Bcl-2) e apoptóticos (citocromo C, a caspase 9, APAF-1), caspase 3 e SMAC/DIABLO, relacionados à apoptose, em 30 amostras de tecido de pacientes com glioblastoma. Métodos: A expressão gênica foi avaliada em trinta glioblastomas e comparada a dez amostras controles de substância branca por PCR em tempo real. Resultados e Conclusão: Houve maior nível de expressão de XIAP (p = 0,0032) e Bcl-2 (p = 0,0351) em comparação com as amostras controle, de cérebro normal. Estes resultados levantam a questão de que os genes Bcl-2 e XIAP podem ser responsáveis pela inibição da morte celular programada em glioblastomas, além disso, proporcionam informação adicional capaz de permitir o desenvolvimento de novas estratégias de terapia alvo.

ABSTRACT Glioblastoma (GBM) is the most malignant glioma and represents 29% of all brain tumors. Tumorigenesis is intimately connected with characteristics acquired in the physiologic pathway of cellular death. Objective: In the present study, the expression of anti-apoptotic (XIAP and Bcl-2) and apoptotic (cytochrome C, caspase 9, APAF-1), caspase 3 and the Smac/DIABLO genes related to the apoptosis pathway were evaluated in 30 samples of glioblastoma. Methods: The gene expression was evaluated in 30 glioblastomas (WHO grade IV) and compared to 10 white matter control samples with real-time PCR. Results and Conclusion: There were higher expressions of XIAP (p = 0.0032) and Bcl-2 (p = 0.0351) in the glioblastoma samples compared to the control samples of normal brain. These results raise the question of whether Bcl-2 and XIAP genes can be responsible for the inhibition of programmed cell death in glioblastomas. Moreover, they provide additional information capable of allowing the development of new target therapy strategies.

https://doi.org/10.1590/0004-282x20170156 1605 downloads

10.

High expression of anti-apoptotic genes in grade I and II meningiomas
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Tirapelli, Daniela Pretti da Cunha

; Menezes, Sarah Bomfim

; Franco, Indira Maynart

; Lustosa, Isis Lacrose

; Rodrigues, Andressa Romualdo

; Novais, Paulo Cézar

; Santiago, Antônio César Mendes

; Peria, Fernanda Maris

; Serafini, Luciano Neder

; Marinho, Alexandre Magno da Nóbrega

; Carlotti Jr, Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luís Fernando

Arquivos de Neuro-Psiquiatria

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apr 2017, Volume 75 N. 4 Pages 209 - 215

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO Um dos diferentes mecanismos genéticos envolvidos na carcinogênese de meningiomas é influenciado por interações entre proteínas que induzem e inibem a apoptose. Objetivos Avaliar a expressão de c-FLIP, XIAP, Bcl-2, caspase 3, 8 e 9, citocromo C, APAF 1 e Smac/DIABLO, genes relacionados com as vias da apoptose. Métodos A expressão gênica foi avaliada em trinta amostras de meningiomas (OMS grau I e II) e em dez amostras normais (de aracnóide) por PCR em tempo real. Resultados Os resultados mostraram maior expressão de genes antiapoptóticos em meningiomas quando comparados com controle, em contraste com a menor expressão de genes próapoptóticos. Conclusão Há um possível bloqueio na ativação de caspases através da via intrínseca da apoptose em meningiomas. O c-FLIP modula a caspase 8 e, desse modo, inibindo a sua ativação pela ausência de ligação com o receptor, há um bloqueio na ativação de FAS pela via extrínseca da apoptose.

ABSTRACT One of the different genetic mechanisms involved in the carcinogenesis of meningiomas is influenced by interactions between proteins that induce and inhibit apoptosis. Objective To evaluate the expression of c-FLIP, XIAP, Bcl-2, caspase 3, 8 and 9, cytochrome c, APAF 1 and Smac/DIABLO genes related to apoptosis pathways. Methods The gene expression was evaluated in 30 meningiomas (WHO grades I and II) and in 10 normal samples (from arachnoid tissue) through PCR-RT. Results The results showed higher expression of anti-apoptotic genes in meningiomas when compared to the control group, which had a low expression of pro-apoptotic genes. Conclusion There is a possible block in the activation of caspases through the intrinsic apoptosis pathway in meningiomas. c-FLIP modulates caspase 8 and, by inhibiting its activation due to the lack of connection with the receiver, there is a block to the FAS activation of apoptosis by its extrinsic pathway.

https://doi.org/10.1590/0004-282x20170027 1533 downloads

11.

Analysis of gene expression EGFR and KRAS, microRNA-21 and microRNA-203 in patients with colon and rectal cancer and correlation with clinical outcome and prognostic factors
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Carvalho, Thais Inácio de

; Novais, Paulo Cezar

; Lizarte Neto, Fermino Sanches

; Sicchieri, Renata Danielle

; Rosa, Marcella Suelma Torrecillas

; Carvalho, Camila Albuquerque Mello de

; Tirapelli, Daniela Pretti da Cunha

; Peria, Fernanda Maris

; Rocha, José Joaquim Ribeiro da

; Féres, Omar

Acta Cirúrgica Brasileira

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mar 2017, Volume 32 N. 3 Pages 243 - 250

Abstract: En | Text: En | PDF: En

Abstract Purpose: To evaluate the expression of EGFR, KRAS genes, microRNAs-21 and 203 in colon and rectal cancer samples, correlated with their age at diagnosis, histological subtype, value of pretreatment CEA, TNM staging and clinical outcome. Methods: Expression of genes and microRNAs by real time PCR in tumor and non-tumor samples obtained from surgical treatment of 50 patients. Results: An increased expression of microRNAs-21 and 203 in tumor samples in relation to non-tumor samples was found. There was no statistically significant difference between the expression of these genes and microRNAs when compared to age at diagnosis and histological subtype. The EGFR gene showed higher expression in relation to the value of CEA diagnosis. The expression of microRNA-203 was progressively lower in relation to the TNM staging and was higher in the patient group in clinical remission. Conclusions: The therapy of colon and rectum tumors based on microRNAs remains under investigation reserving huge potential for future applications and clinical interventions in conjunction with existing therapies. We expect, based on the exposed data, to stimulate the development of new therapeutic possibilities, making the treatment of these tumors more effective.

https://doi.org/10.1590/s0102-865020170030000009 1908 downloads

12.

Expression profiles of eNOS, iNOS and microRNA-27b in the corpus cavernosum of rats submitted to chronic alcoholism and Diabetes mellitus
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Cunha, Joao Paulo da

; Lizarte Neto, Fermino Sanches

; Novais, Paulo Cezar

; Gattas, Daniela

; Carvalho, Camila Albuquerque Mello de

; Tirapelli, Daniela Pretti da Cunha

; Molina, Carlos Augusto Fernandes

; Tirapelli, Luis Fernando

; Tucci Junior, Silvio

Acta Cirúrgica Brasileira

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jan 2017, Volume 32 N. 1 Pages 38 - 45

Abstract: En | Text: En | PDF: En

Abstract Purpose: To evaluate the expression of endothelial and inducible NOS in addition to the miRNA-27b in the corpus cavernosum and peripheral blood of healthy rats, diabetic rats, alcoholic rats and rats with both pathologies. Methods: Forty eight Wistar rats were divided into four groups: control (C), alcoholic (A), diabetic (D) and alcoholic-diabetic (AD). Samples of the corpus cavernosum were prepared to study protein expressions of eNOS and iNOS by immunohistochemistry and expression of miRNA-27b in the corpus cavernosum and peripheral blood. Results: Immunohistochemistry for eNOS and iNOS showed an increase in cavernosal smooth muscle cells in the alcoholic, diabetic and alcoholic-diabetic groups when compared with the control group. Similarly, the mRNA levels for eNOS were increased in cavernosal smooth muscle (CSM) in the alcoholic, diabetic and alcoholic-diabetic groups and miRNA-27b were decreased in CSM in the alcoholic, diabetic and alcoholic-diabetic groups. Conclusion: The major new finding of our study was an impairment of relaxation of cavernosal smooth muscle in alcoholic, diabetic, and alcoholic-diabetic rats that involved a decrease in the nitric oxide pathway by endothelium-dependent mechanisms accompanied by a change in the corpus cavernosum contractile sensitivity.

https://doi.org/10.1590/s0102-865020170105 1446 downloads

13.

Morphological and immunohistochemical analysis of apoptosis in the cerebellum of rats subjected to focal cerebral ischemia with or without alcoholism model
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Carvalho, Camila Albuquerque Melo de

; Tirapelli, Daniela Pretti da Cunha

; Rodrigues, Andressa Romualdo

; Lizarte Neto, Fermino Sanches

; Novais, Paulo Cézar

; Silva, Jairo Pinheiro

; Carlotti Júnior, Carlos Gilberto

; Colli, Benedicto Oscar

; Tirapelli, Luís Fernando

Acta Cirúrgica Brasileira

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sep 2016, Volume 31 N. 9 Pages 629 - 637

Abstract: En | Text: En | PDF: En

ABSTRACT PURPOSE: To evaluated histopathological changes, morphometric and expression of proteins CASPASE-3, BCL-2 and XIAP related to apoptosis in the cerebellum after induction of temporary focal cerebral ischemia followed by reperfusion, with or without a model of chronic alcoholism. METHODS: Fifty Wistar rats were used and divided into: control group (C), sham group (S), ischemic group (I), alcoholic group (A), and ischemic and alcoholic group (IA). The cerebellum samples collected were stained for histopathological and morphometric analysis and immunohistochemistry study. RESULTS: Histopathological changes were observed a greater degree in animals in groups A and IA. The morphometric study showed no difference in the amount of cells in the granular layer of the cerebellum between the groups. The expression of CASPASE-3 was higher than BCL-2 and XIAP in the groups A and IA. CONCLUSION: We observed correlation between histopathological changes and the occurrence of apoptosis in cerebellar cortex.

https://doi.org/10.1590/S0102-865020160090000009 2227 downloads

14.

Comparative evaluation of oncologic outcomes in colon cancer
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Bernardes, Mário Vinícius Angelete Alvarez

; Feitosa, Marley Ribeiro

; Peria, Fernanda Maris

; Tirapelli, Daniela Pretti da Cunha

; Rocha, José Joaquim Ribeiro da

; Feres, Omar

Acta Cirúrgica Brasileira

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2016, Volume 31 Suppl. 1 Pages 34 - 39

Abstract: En | Text: En | PDF: En

PURPOSE: In this paper we report clinical variables on colon cancer series. Oncological outcomes were compared to low-income and high-income countries. METHODS: We analysed a prospective database of 51 colon cancer patients submitted to primary tumor resection between 2010 and 2011, showing clinical variables and oncologic outcomes. RESULTS: R0 resection obtained in 80.4%, 21.6% of patients was TNM stage IV, and only 13.7% showed TNM stage I. Disease-free survival was 32 months, overall survival was 46 months, and the tumoral recurrence rate was 9.8%. Univariate analysis showed association of serum CEA levels ≥ 5 ng/dl (p= 0.004), presence of metastasis at diagnosis (p= 0.012), compromised surgical margins (p < 0.001) and poorer tumor differentiation (p= 0.041) to death. Multivariate analysis identified compromised surgical margins as an independent risk factor for death due to colon cancer (P=0.003; odds ratio=0.36; 95% confidence interval=0.004-0.33). Nowadays, 62.7% of patients are alive. CONCLUSION: Recurrence rate, disease-free survival and overall survival was similar to those observed in more developed countries. Serum CEA levels ≥ 5 ng/dl, the presence of metastasis at diagnosis, compromised surgical margins and poorer tumor differentiation were associated with death. A compromised surgical margin was the only independent risk factor for death.

https://doi.org/10.1590/S0102-86502016001300008 2005 downloads

15.

Serological under expression of microRNA-21, microRNA-34a and microRNA-126 in colorectal cancer
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Almeida, Ana Luiza Normanha Ribeiro de

; Bernardes, Mário Vinícius Angelete Alvarez

; Feitosa, Marley Ribeiro

; Peria, Fernanda Maris

; Tirapelli, Daniela Pretti da Cunha

; Rocha, José Joaquim Ribeiro da

; Feres, Omar

Acta Cirúrgica Brasileira

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2016, Volume 31 Suppl. 1 Pages 13 - 18

Abstract: En | Text: En | PDF: En

PURPOSE: This paper describes the ability of miRNA value predict oncological outcomes in CRC patients and correlates to clinical and pathologic variables. METHODS: We prospectively analyzed the serological expression of microRNA-21, microRNA-34a, and microRNA-126 in 37 stage II - IV CRC patients and correlate to seven fit counterparts. Serological microRNAs were extracted using the miRNeasy Mini Kit(r) (Qiagen, Hilden, Germany). Quantification of microRNAs was performed using TaqMan Master Mix(r) reagent (Applied Biosystems, USA). RESULTS: We obtained serological underexpression microRNA-21, microRNA-34a, and microRNA-126 in CRC group. However, miRNAs serological values do not impact prognosis. Furthermore, miRNAs was not influenced by CEA values, TNM staging, and histological subtype. CONCLUSION: Despite lower expression of miR-21, miR-34a and miR-126 in the CRC group, no association with poor prognosis was found.

https://doi.org/10.1590/S0102-86502016001300004 2615 downloads

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