Meningeome sind Tumore der Hirnhäute und stellen zirka 24-30% aller intrakraniellen Tumore dar. Obwohl sie in den meisten Fällen als solitär, langsam wachsend und benigne (WHO Grad 1) beschrieben werden, ist ihr ausgeprägtes Rezidivverhalten die größte Herausforderung in der Therapie. Bisherige Arbeiten verwendeten zur genetischen Analyse von Meningeomen meist Untersuchungstechniken mit eingeschränkter (molekular-)zytogenetischer Aussagekraft. Mit der Kombination der Methoden Giemsa-Bandendarstellung (GTG-Bänderung), Spektrale Karyotypisierung (SKY-Technik), Fluoreszenz in situ Hybridisierungstechniken (FISH-Analyse) und molekulare Karyotypisierung unter Verwendung von 100K beziehungsweise 6.0 SNP-Arrays (SNP-A Karyotypisierung) sollte es möglich sein, in effizienterer Form bislang unentdeckte chromosomale Aberrationen zu identifizieren und weiterführende tumormechanistische Hinweise zu erhalten. In der vorliegenden Arbeit wurde zunächst ein multipel aufgetretenes Meningeom mit zwei Tumoren unterschiedlicher Malignität (1 WHO Grad 1; 1 WHO Grad 2) analysiert, anschließend erfolgte die Untersuchung einer Gruppe von 10 Meningeomen (5 WHO Grad 1; 5 WHO Grad 2). Bisher nicht beschriebene Aberrationen wie ein dizentrisches Chromosomen 4, die parazentrische Inversion im chromosomalen Bereich 1p36 und die balancierte reziproke Translokation t(4;10)(q12;q26) wurden detektiert. Die genomweite SNP-A Karyotypisierung ermöglichte neben der genaueren Betrachtung der zytogenetischen Ergebnisse die simultane Analyse von Blut und Tumor-DNA der Patienten und lieferte Hinweise auf konstitutionelle Aberrationen. Es zeigte sich eine signifikante Anreicherung von rekurrenten Regionen kopienneutraler Verluste der Heterozygotie als Hinweis auf das Vorliegen potenzieller segmentaler Uniparentaler Disomie (UPD) jeweils in Blut und Tumor der Patienten. Außerdem wurden nur im Tumor befindliche potentielle rekurrente segmentale UPD Regionen detektiert. Die weitere Analyse der konstitutionellen sowie somatischen segmentalen UPD hinsichtlich ihrer Rolle im Rahmen der Tumorgenese ist eine wichtige Aufgabe für die Zukunft.

Purpose: Meningiomas are common intracranial tumors, often well controlled with surgical resection alone. While the efficacy of radiation therapy in improving local control and progression-free survival is well documented, prognostic data substantiate factors that are predictive of poor local control following definitive radiation therapy. PCNA is a DNA polymerase expressed at the highest levels in the S-phase, the most resistant portion of the cell cycle to ionizing radiation in vitro. We investigated the possible correlation between the levels of PCNA expression and the clinical outcome of patients treated with definitive radiation therapy. Methods and Materials: Archival tissue was collected from 33 cases of meningioma treated at our institution for definitive radiation therapy between 1970 and 1990. Age-matched normal meningeal tissue and asymptomatic meningiomas removed at autopsy served as tissue controls. A standard ABC immumoperoxidase technique employing antibodies to PCNA, PC-10 (Dako, California) was used to stain specimen slides for PCNA. PCNA index was defined as the number of positive nuclei per 10 high-power fields at 400x magnification. Two independent observers scored the slides without prior knowledge of the cases at hand. Results: Patients with high PCNA index were less likely to be controlled by therapeutic radiation (p < 0.001, Kaplan- Meier). All patients with a PCNA index greater that 25 failed radiation therapy. Using multivariate analyses, malignant (but not atypical), histology and PCNA index were significant predictors of progression following radiation therapy (p < 0.05, log rank). Conclusion: PCNA index may be a useful adjunct to more standard histopathologic criteria in the determination of meningioma local control and progression-free survival following therapeutic irradiation. Data on a more expanded population evaluated on a prospective basis will be needed before such criteria are routinely employed in the clinical setting.

No description available.

Introdução: Radiocirurgia estereotática (RCE) e radioterapia estereotática fracionada (RCEF) são inovações modernas de procedimentos radioterápicos, de alta precisão que modelam o feixe de radiação para coincidir com o contorno da lesão, por meio de um sistema de imobilização exata do paciente ao aparelho, com definição do alvo através da fusão de imagens de RM, TC, Angiografia e PET/CT; em que pelas coordenadas de referência estereotática, determina-se que a dose de radiação de alta energia prescrita pelo médico seja depositada somente no volume-alvo, com preservação dos tecidos sadios, órgãos ou estruturas localizadas em suas adjacências. Meningeomas do seio cavernoso (MSCs) representam um problema especial porque podem evoluir comprimindo ou infiltrando estruturas neurovasculares presentes no seio cavernoso. Há evidências de que a RCE e a RCEF proporcionam controle satisfatório do crescimento dos meningeomas do seio cavernoso (MSCs) com efeitos adversos reduzidos. Objetivo: Avaliar resultados da avaliação clínica e da neuroimagem de doentes sintomáticos com MSCs tratados com RCEF ou RCE exclusivamente ou de modo adjuvante à neurocirurgia. Casuística e métodos: Estudo tipo coorte e retrospectivo sobre a avaliação de 89 doentes com MSC sintomático tratados com RCE (36%) ou RCEF (64%) entre janeiro de 1994 e março de 2009 e acompanhados até o final de 2012. Haviam sido submetidos à ressecação neurocirúrgica parcial (Simpson IV) ou à biopsia (Simpson V) previamente à radioterapia 29,2% dos doentes. A dose média de RCE foi de 14Gy, e a dose total de RCEF variou entre 50,4 e 54Gy, sendo fracionada em 1,8-2Gy/dose/dia. Resultados: O período de acompanhamento variou entre 36 e 180 meses (mediana de 73 meses). A percentagem de melhora dos sintomas neuroclínicos individuais e de melhora clínica e radiológica (p > 0,05) apresentou valores semelhantes nos doentes tratados com RCE ou RCEF, sendo respectivamente de 41,6% e 48,3%. Em 37% dos doentes, houve manutenção de, pelo menos, uma queixa neurológica apresentada antes do tratamento e, em 43,8%, a imagem do MSC manteve-se inalterada. O período livre de progressão do MSC em 5, 10 e 15 anos foi de 98,8%, 92,3% e 92,3%, respectivamente. Houve progressão da doença em quatro doentes (4,5%). A melhora dos sintomas neurológicos em doentes submetidos previamente à neurocirurgia ocorreu de maneira mais lenta em relação aos não operados, em razão de manipulação de nervos cranianos. Alguns sintomas pós-operatórios, como a ptose palpebral unilateral, persistiram permanentemente. Nenhuma complicação grave foi observada. Sete doentes apresentaram neuropatia óptica transitória durante 3 meses que melhorou com o uso de corticoides, dois neuropatia trigeminal que melhorou com uso de esteroides e um doente apresentou obstrução total da artéria carótida interna sem repercussão neurológica. Letargia e cefaléia foram os sintomas temporários imediatos mais frequentes. Conclusões: A RCEF e a RCE são métodos seguros e eficazes para tratar doentes com MSC sintomático. Proporcionam melhora ou estabilização da sintomatologia na maioria dos casos, e estabilização ou regressão do tumor demonstrado pela neuroimagem em mais de 90% deles. Ocorreu recuperação dos sintomas neurológicos preexistentes mais rapidamente em doentes não submetidos previamente à neurocirurgia. Houve recorrência do tumor em 4,5% dos doentes. Em até 15 anos de acompanhamento, não se evidenciou indução tumoral com o tratamento / Introduction: Stereotactic radiosurgery (SRS) and fractionated stereotactic radiotherapy (FSRS) are modern innovations in radiotherapy procedures, precision shaping the radiation beam to match the contour of the lesion, through a system of accurate patient immobilization to the device, defining target through the fusion of MRI, CT, angiography and PET / CT, which is determined by reference to stereotactic coordinates. The radiation dose of high energy prescribed by the doctor to be delivery only in the target interest, with preservation of healthy tissues, organs or structures located in their vicinity. Cavernous sinus meningiomas (CSMs) pose a special problem because they can evolve compressing or infiltrating the neurovascular structures present of the cavernous sinus. There are evidences that SRS and FRS are efficient in the treatment of CSMs. Objectives: The evaluation of the long-term clinical results and neuroimaging findings in patients with symptomatic CSM treated with FSRT or SRS as single therapy or after a previous neurosurgical treatment. Patients and methods: Retrospective cohort study involving 89 patients with symptomatic CSMs treated with SRS (36%) or FSRS (64%) from January 1994 to March 2009, and followed until the end of 2012. Previous neurosurgical partial resection (Simpson IV) or biopsies (Simpson V) had been performed in 29.2% of the patients. The median dose of SRS was 14Gy and the total dose of FSRT ranged from 50.4 to 54Gy, fractionated in 1.8 to 2Gy/dose/day. Results: The follow-up period ranged from 36 to 180 months (median= 73months). There was improvement in the individual symptoms and in the clinical and radiological findings regardless the radiotherapeutic method in 41.6% and 48.3% of the patients treated with SRS or FSRT, respectively (p > 0,05). In 37% of the patients, at least one neurological complaint present before the treatment did not change and in 43.8% patients, the image of the tumor remained stable. The progression-free survival in 5, 10 and 15 years was 98.8%, 92.3% and 92.3%, respectively. The improvement of neurological symptoms in patients previously treated with neurosurgery was slower or did not occur as in nonpreviously operated patients. Lethargy and headache were the most frequent transient immediate post-radiotherapy symptoms. Seven patients presented transient optic neuropathy during 3 months and improved with corticosteroids, 2 presented trigeminal neuropathy that remitted rapidly with steroids, and one, had total occlusion of the internal carotid artery without neurological consequences. Conclusions: Both FSRT and SRS were equally safe and effective in the management of symptomatic CSMs. There was improvement or stabilization of the neurological symptoms in the majority ofthe patients and stabilization or regression of the neuroimaging of the lesion in more them 90% of them. The recovery of preexisting cranial neuropathies occurred faster and was more frequent in patients not previously treated with surgical procedure. There was recurrence in 4.5% of the patients. No radiation-induced tumor was observed term during the longest 15 years follow-up

Introdução: Radiocirurgia estereotática (RCE) e radioterapia estereotática fracionada (RCEF) são inovações modernas de procedimentos radioterápicos, de alta precisão que modelam o feixe de radiação para coincidir com o contorno da lesão, por meio de um sistema de imobilização exata do paciente ao aparelho, com definição do alvo através da fusão de imagens de RM, TC, Angiografia e PET/CT; em que pelas coordenadas de referência estereotática, determina-se que a dose de radiação de alta energia prescrita pelo médico seja depositada somente no volume-alvo, com preservação dos tecidos sadios, órgãos ou estruturas localizadas em suas adjacências. Meningeomas do seio cavernoso (MSCs) representam um problema especial porque podem evoluir comprimindo ou infiltrando estruturas neurovasculares presentes no seio cavernoso. Há evidências de que a RCE e a RCEF proporcionam controle satisfatório do crescimento dos meningeomas do seio cavernoso (MSCs) com efeitos adversos reduzidos. Objetivo: Avaliar resultados da avaliação clínica e da neuroimagem de doentes sintomáticos com MSCs tratados com RCEF ou RCE exclusivamente ou de modo adjuvante à neurocirurgia. Casuística e métodos: Estudo tipo coorte e retrospectivo sobre a avaliação de 89 doentes com MSC sintomático tratados com RCE (36%) ou RCEF (64%) entre janeiro de 1994 e março de 2009 e acompanhados até o final de 2012. Haviam sido submetidos à ressecação neurocirúrgica parcial (Simpson IV) ou à biopsia (Simpson V) previamente à radioterapia 29,2% dos doentes. A dose média de RCE foi de 14Gy, e a dose total de RCEF variou entre 50,4 e 54Gy, sendo fracionada em 1,8-2Gy/dose/dia. Resultados: O período de acompanhamento variou entre 36 e 180 meses (mediana de 73 meses). A percentagem de melhora dos sintomas neuroclínicos individuais e de melhora clínica e radiológica (p > 0,05) apresentou valores semelhantes nos doentes tratados com RCE ou RCEF, sendo respectivamente de 41,6% e 48,3%. Em 37% dos doentes, houve manutenção de, pelo menos, uma queixa neurológica apresentada antes do tratamento e, em 43,8%, a imagem do MSC manteve-se inalterada. O período livre de progressão do MSC em 5, 10 e 15 anos foi de 98,8%, 92,3% e 92,3%, respectivamente. Houve progressão da doença em quatro doentes (4,5%). A melhora dos sintomas neurológicos em doentes submetidos previamente à neurocirurgia ocorreu de maneira mais lenta em relação aos não operados, em razão de manipulação de nervos cranianos. Alguns sintomas pós-operatórios, como a ptose palpebral unilateral, persistiram permanentemente. Nenhuma complicação grave foi observada. Sete doentes apresentaram neuropatia óptica transitória durante 3 meses que melhorou com o uso de corticoides, dois neuropatia trigeminal que melhorou com uso de esteroides e um doente apresentou obstrução total da artéria carótida interna sem repercussão neurológica. Letargia e cefaléia foram os sintomas temporários imediatos mais frequentes. Conclusões: A RCEF e a RCE são métodos seguros e eficazes para tratar doentes com MSC sintomático. Proporcionam melhora ou estabilização da sintomatologia na maioria dos casos, e estabilização ou regressão do tumor demonstrado pela neuroimagem em mais de 90% deles. Ocorreu recuperação dos sintomas neurológicos preexistentes mais rapidamente em doentes não submetidos previamente à neurocirurgia. Houve recorrência do tumor em 4,5% dos doentes. Em até 15 anos de acompanhamento, não se evidenciou indução tumoral com o tratamento / Introduction: Stereotactic radiosurgery (SRS) and fractionated stereotactic radiotherapy (FSRS) are modern innovations in radiotherapy procedures, precision shaping the radiation beam to match the contour of the lesion, through a system of accurate patient immobilization to the device, defining target through the fusion of MRI, CT, angiography and PET / CT, which is determined by reference to stereotactic coordinates. The radiation dose of high energy prescribed by the doctor to be delivery only in the target interest, with preservation of healthy tissues, organs or structures located in their vicinity. Cavernous sinus meningiomas (CSMs) pose a special problem because they can evolve compressing or infiltrating the neurovascular structures present of the cavernous sinus. There are evidences that SRS and FRS are efficient in the treatment of CSMs. Objectives: The evaluation of the long-term clinical results and neuroimaging findings in patients with symptomatic CSM treated with FSRT or SRS as single therapy or after a previous neurosurgical treatment. Patients and methods: Retrospective cohort study involving 89 patients with symptomatic CSMs treated with SRS (36%) or FSRS (64%) from January 1994 to March 2009, and followed until the end of 2012. Previous neurosurgical partial resection (Simpson IV) or biopsies (Simpson V) had been performed in 29.2% of the patients. The median dose of SRS was 14Gy and the total dose of FSRT ranged from 50.4 to 54Gy, fractionated in 1.8 to 2Gy/dose/day. Results: The follow-up period ranged from 36 to 180 months (median= 73months). There was improvement in the individual symptoms and in the clinical and radiological findings regardless the radiotherapeutic method in 41.6% and 48.3% of the patients treated with SRS or FSRT, respectively (p > 0,05). In 37% of the patients, at least one neurological complaint present before the treatment did not change and in 43.8% patients, the image of the tumor remained stable. The progression-free survival in 5, 10 and 15 years was 98.8%, 92.3% and 92.3%, respectively. The improvement of neurological symptoms in patients previously treated with neurosurgery was slower or did not occur as in nonpreviously operated patients. Lethargy and headache were the most frequent transient immediate post-radiotherapy symptoms. Seven patients presented transient optic neuropathy during 3 months and improved with corticosteroids, 2 presented trigeminal neuropathy that remitted rapidly with steroids, and one, had total occlusion of the internal carotid artery without neurological consequences. Conclusions: Both FSRT and SRS were equally safe and effective in the management of symptomatic CSMs. There was improvement or stabilization of the neurological symptoms in the majority ofthe patients and stabilization or regression of the neuroimaging of the lesion in more them 90% of them. The recovery of preexisting cranial neuropathies occurred faster and was more frequent in patients not previously treated with surgical procedure. There was recurrence in 4.5% of the patients. No radiation-induced tumor was observed term during the longest 15 years follow-up

Introduction et objectifs : Les méningiomes sont des tumeurs intracrâniennes pour la plupart de bas grade, dont les récidives sont cependant fréquentes. Le premier objectif de cette étude est d'évaluer la valeur pronostique de marqueurs de prolifération dans une série de 60 méningiomes. HuR (ELAV-like 1) est une protéine pro-oncogène pouvant stimuler la prolifération et la survie cellulaire. Dans la seconde partie de cette étude, nous analysons l'expression de HuR et les effets de l'inhibition de HuR sur la prolifération, l'apoptose et la résistance à l'hypoxie. Résultats : Il existe une corrélation significative entre le grade histologique et l'expression nucléaire de MCM6 (p<0,001), Ki-67 (p<0,001) et PHH3 (p<0,001), et une corrélation inverse entre ces marqueurs et la survie sans récidive (Cox ; MCM6 : p<0,001 ; Ki-67 : p=0,003 ; PHH3 : p=0,037). L'expression cytoplasmique de HuR est corrélée négativement avec la survie sans récidive (p=0,028), et positivement avec le grade (I vs II : p=0,0007), l'indice mitotique (p=0,0001), Ki-67 (p=0,0007) et MCM6 (p=0,0009). In vitro, l'inhibition de HuR (siRNA) entraîne une diminution de la croissance cellulaire (p=0,0004) et de la prolifération (Ki-67, p=0,0004), et une augmentation de l'apoptose (caspase 3 clivée, p=0,002), ces différents effets étant significativement augmentés en hypoxie. L'inhibition de HuR entraîne également une diminution de SIRT1 en normoxie (p=0,01) et en hypoxie (p=0,0006). Conclusion : MCM6 est un marqueur efficace pour identifier les méningiomes à haut risque de récidive. HuR est un marqueur de mauvais pronostic, dont l'inhibition permet de diminuer la prolifération cellulaire et d'augmenter l'apoptose / Introduction and objectives: Meningiomas are frequent and in most of cases low grade intracranial tumors, with frequent recurrences. The first objective of this study was to evaluate the prognostic value of proliferation and cell cycle markers in a series of 60 meningiomas. HuR (ELAV-like 1) is a pro-oncogenic protein that stimulates cell proliferation and survival. In the second part of this study, the objective was to evaluate HuR expression, and the effects of HuR inhibition on proliferation, apoptosis and resistance to hypoxia. Results: We found a significant correlation between histological grade and nuclear staining for MCM6 (p<0.001), Ki-67 (p<0.001) and PHH3 (p<0.001), and an inverse correlation between these markers and reccurence free survival (Cox; MCM6: p<0.001; Ki-67: p=0.003; PHH3: p=0.037). Cytoplasm staining for HuR was significantly stronger in atypical meningiomas (grades I vs II: p=0.0007), inversely correlated with progression free survival (p=0.028), and was positively correlated with mitotic index (p=0.0001), Ki-67 (p=0.0007) and MCM6 (p=0.0009). In vitro, HuR inhibition (siRNA) led to a significant decrease of cell growth (p=0.0004) and proliferation (Ki-67, p=0.0004), and an increase of apoptosis (cleaved caspase 3, p=0.002). These effects were significantly increased under hypoxia, comparing with normoxia. When HuR was inhibited, SIRT1 was decreased, both under normoxia (p=0.01) and hypoxia (p=0.0006). Conclusion: In conclusion, MCM6 is an efficient marker to identify meningiomas with high risk of recurrence. HuR is correlated with a poor prognosis. Its inhibition allows to decrease cell proliferation and to increase apoptosis

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No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Tese_AlteracoesGeneticasEpigeneticas.pdf: 1341053 bytes, checksum: 3c3dc3d55f7d58401af456f5fcaabb7c (MD5) Previous issue date: 2013 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / FAPESPA - Fundação Amazônia de Amparo a Estudos e Pesquisas / Os meningiomas são os tumores intracranianos mais frequentes, originando-se das meninges que revestem o cérebro e cordão espinhal. Apesar de terem sido um dos primeiros neoplasmas sólidos estudados citogeneticamente, ainda são escassos os estudos genéticos e epigenéticos nesses tumores. O presente trabalho teve como objetivo investigar alterações genéticas e epigenéticas que pudessem contribuir na iniciação e progressão tumoral em meningiomas na população paraense. Essa tese está subdivida em três capítulos. No Capítulo I foi investigada a associação entre o polimorfismo MTHFR C677T e meningioma em 23 pacientes da população paraense, utilizando 96 indivíduos sem histórico de lesões pré-neoplásicas como grupo controle. Essa associação não foi encontrada, apesar de sugerir um aumento não estatisticamente significante no risco de desenvolver meningioma em portadores do genótipo TT quando comparados ao genótipo CC. No Capítulo II foi avaliado o padrão de metilação em duas famílias do microRNA124 em meningiomas na população paraense. Hipermetilação na região promotora de miRN124a2 e miRNA124a3 parece ser um evento frequente, uma vez que foi encontrada em 73,9% e 69,56% das amostras analisadas, respectivamente. No Capítulo III, foi analisado o padrão de metilação dos genes APC, BRCA1, CDH1, CDH13, CDKN2A, DAPK1, ESR1, FHIT, GSTP1, MGMT, MLH1, NEUROG1, PDLIM4, PTEN, RARB, RASSF1, RUNX3, SOCS1, TIMP3, TP73, VHL e WIF1 em um meningioma de grau I e um de grau II através de uma placa comercial desenvolvida através da tecnologia MethylScreen. O padrão de metilação do gene CDKN2B também foi analisado na amostra coletada em 25 pacientes com meningioma através da conversão por bissulfito, PCR e sequenciamento direto. O gene RASSF1A apresentou-se metilado em 16,73% e 63,66% dos sítios CpGs analisados na amostra de meningioma de grau I e grau II, respectivamente. O gene RUNX3 se apresentou metilado apenas na amostra de grau II em 52,88% dos sítios CpG analisados. Nossos resultados apontam a importância das alterações epigenéticas na tumorigênese e progressão tumoral em meningiomas. / Meningiomas are the most common intracranial tumors that originate from the meninges surrounding the brain and spinal cord. Despite meningiomas were among the first solid neoplasms to be studied cytogenetically, little is known about their genetic and epigenetic profile. This study aimed to investigate genetic and epigenetic alterations that could contribute to tumor initiation and progression in meningiomas in the population of Pará, Brazil. This thesis is subdivided into three chapters. In Chapter I we investigated the association between the MTHFR C677T and meningioma in 23 patients in the population of Pará. A total of 96 healthy individuals with no previous pre-neoplastic lesions were selected for the control group. This association was not found. Although not statistically significant, our observation suggests that the TT genotype increases the risk of developing meningioma when compared to CC genotype. In Chapter II we evaluated the methylation pattern in two members of microRNA124 family in meningiomas in the population of Pará. Hypermethylation of the promoter region of miRN124a2 and miRNA124a3 appears to be a frequent event, as was found in 73.9% and 69.56% of the samples, respectively. In Chapter III, we analyzed the methylation pattern of the APC, BRCA1, CDH1, CDH13, CDKN2A, DAPK1, ESR1, FHIT, GSTP1, MGMT, MLH1, NEUROG1, PDLIM4, PTEN, Rb, RASSF1, RUNX3, SOCS1, TIMP3, TP73, VHL and WIF1 genes in a grade I and in a grade II meningiomas through an assay developed by MethylScreen. Pattern of methylation of CDKN2B was also analyzed in 25 patients with meningioma through bisulfite conversion, PCR and direct sequencing. RASSF1A was methylated in 16.73% and 63.66% of the CpG sites analyzed in the grade I and grade II meningioma, respectively. RUNX3 is methylated only in grade II meningioma in 52.88% of the CpG sites analyzed. Our results point to the importance of epigenetic changes in tumorigenesis and tumor progression in meningiomas.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-03-27T13:33:17Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AnaliseGenesTP53.pdf: 906769 bytes, checksum: 868736765e91f437e42d5aca6b356441 (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-03-30T12:15:36Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AnaliseGenesTP53.pdf: 906769 bytes, checksum: 868736765e91f437e42d5aca6b356441 (MD5) / Made available in DSpace on 2017-03-30T12:15:36Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AnaliseGenesTP53.pdf: 906769 bytes, checksum: 868736765e91f437e42d5aca6b356441 (MD5) Previous issue date: 2016-06-17 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / Apesar da considerável incidência, estudos de alterações genéticas nos genes TP53, PTEN, IDH1 e IDH2, em tumores não gliais, são raros e, em alguns casos, inexistentes. Os tumores não gliais são classificados geralmente como benignos e raramente evoluem à malignidade, apresentando diferentes classificações, incidências e localizações. Os genes supressores tumorais e de resposta a danos ao DNA, TP53 e PTEN, estão entre os genes mais frequentemente mutados em tumores humanos. Os genes IDH1 e IDH2 estão envolvidos no metabolismo celular e, também, foram encontrados frequentemente mutados em gliomas, melanomas e leucemias, sendo atualmente considerados como bons marcadores em gliomas. Foram realizadas análises de alterações genéticas nos genes citados, a fim de verificar se estão associados à etiologia e/ou progressão de tumores não gliais do Sistema Nervoso Humano (SNH). Foram utilizadas as técnicas de PCR-SSCP para amplificação da região de interesse e triagem mutacional das amostras para posterior sequenciamento. Foram analisadas 37 amostras de tumores não gliais (14 schwannomas, 3 Meningiomas, 4 Meduloblastomas, 2 Neurocitomas e 14 Metástases do Sistema Nervoso Central (SNC). Somente o gene IDH1 apresentou polimorfismos na SSCP em 12 (32,4%) amostras, sendo, então, submetidas ao sequenciamento. No entanto, as reações de sequenciamento foram satisfatórias em apenas em 5 amostras, entre as polimórficas, (1 metástase, 1 meningioma e 3 schwanomas,). Análises dessas 5 amostras identificaram diferentes mutações, uma delas, presente em todas, uma transversão T→A no éxon 4 do códon 106 do gene IDH1, resultando na substituição do aminoácido treonina por serina. Foram, também, identificadas outras mutações em regiões não codificantes (íntron 4) do gene IDH1 em duas dessas amostras. As mutações encontradas em nosso estudo ainda não haviam sido relatadas na literatura. Nossos resultados indicam a participação do gene IDH1 na patogênese desses tumores. / Despite the considerable incidence, studies of genetic changes in gene TP53, PTEN, IDH2 and IDH1, in not glial tumors are rare and, in some cases, nonexistent. Glial tumors are usually not classified as benign and rarely evolve to malignancy, with different classifications, effects and locations. The tumor suppressor genes and response to DNA damage, TP53 and PTEN are among the most commonly mutated gene in human tumors. The genes IDH1 and IDH2 are involved in cell metabolism and also were frequently found mutated in gliomas, melanomas and leukemias, currently being considered as good markers for gliomas. Genetic analyzes were performed in those genes, in order to verify that are associated with the etiology and/or progression of non-glial tumors of Human Nervous System (HNS). SSCPPCR techniques for the amplification of the region of interest and mutational screening of samples for subsequent sequencing were used. We analyzed 37 samples of non-glial tumors (14 schwannomas, meningiomas 3, 4 Medulloblastomas, 2 neurocytomas and 14 metastases of Central Nervous System (CNS). Only the gene IDH1 polymorphisms presented on the SSCP 12 (32.4%) samples, and then subjected to sequencing. However, sequencing reactions were satisfactory in only 5 samples, of the polymorphic, (1 metastasis, meningioma 1 and 3 schwannomas). Analysis of these samples have identified 5 different mutations, one present in all, one transversion T → A in codon 106 of exon 4 of the IDH1 gene resulting in amino acid substitution of threonine by serine. Were also identified other mutations in noncoding regions (intron 4) of gene IDH1 in two of these samples. The mutations found in our study had not yet been reported in the literature. Our results indicate the participation the gene IDH1 in the pathogenesis of these tumors.

Malignant gliomas are the most common primary brain tumors in adults. Patients with these highly malignant tumors have an extremely poor prognosis. The situation with a highly proliferative tumor in a non-proliferating tissue should favor cytostatic treatment but so far the role of conventional chemotherapy has been adjunctive. The concentrations of three sex steroids, estradiol, progesterone and testosterone, were analyzed by radioimmunoassay after celite chromatography in brain tumor samples. Some malignant gliomas had high tissue concentrations of estradiol. Low progesterone levels may suggest steroid consumption. Estramustine (EM), a conjugate of estradiol-17ß and nornitrogen mustard had a dose-dependent antiproliferative effect on several human malignant glioma cell lines. At equimolar concentrations the inhibitory effects of the EM complex were clearly more pronounced than those of estradiol and nornitrogen mustard given alone or in combination. A specific binding protein (EMBP) is important for the cytotoxic action of EM. Using a mouse monoclonal antibody and an indirect antibodyperoxidase technique, EMBP was demonstrated in human glioma cells. Significant amounts of EMBP were also detected in human brain tumor tissue by radioimmunoassay. The mean concentrations (ng/mg protein) in 16 astrocytomas (2.6) and 7 meningiomas (5.1) were higher (p&lt;0.001) than in 18 samples of normal brain (0.5). The presence of the specific binding protein may suggest a selective binding and effect of EM in human brain tumor tissue. Human glioma cells displayed significant uptake, retention and metabolism of estramustine phosphate (EMP). After incubation with ^H-EMP a progressive uptake of radioactivity was recorded during 24 hours. Metabolism of parent EMP into estramustine and estromustine, which is a well known part of the metabolic pathway in man, was also demonstrated. A dose- dependent increase in DNA strand breaks was recorded at EMP- concentrations ranging 10-40 yg/ml. The uptake of ®^Rb, used as a tracer for potassium to study ion transport and membrane permeability, was reduced after incubation with EMP. Scanning electron microscopy gave further evidence for membrane damage. According to flow cytometric analyses exponentially growing glioma cells were accumulated in the G2/M stage and the fraction of Gi/Gq was reduced. EM seems to attack malignant cells in a multifocal fashion on several vital functions including the microtubule, the nucleus, and the cell membrane. The intact EM complex may be important for effects related to microtubule function which add to the cytotoxic potential of its constituents. These experimental findings justify further investigations on the role of sex hormones in brain tumor growth and development and of hormone-linked cytostatics in clinical treatment. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1990, härtill 6 uppsatser.</p> / digitalisering@umu

Abstract The findings on depression and anxiety among brain tumor patients have so far been based on case series and case samples. In Finland, psychiatric research in relation to psychiatric symptoms among patients with different types of brain tumors is lacking. The study population of this thesis consisted of 101 patients (39 males and 62 females) aged between 20 and 82 years with a solitary primary brain tumor treated surgically at the Oulu Clinic for Neurosurgery, Oulu University Hospital between February 1990 and March 1992. The major histological subgroup consisted of gliomas (40%), and the rest were meningiomas (33%), acoustic neurinomas (13%), pituitary adenomas (8%) and other types (6%). The psychiatric symptoms of the patients were assessed at three time points, namely before tumor operation as well as at three months and at one year after operation by two valid measurement instruments, the Beck Depression Inventory and the Crown Crisp Experiential Index. In addition, the patients' functional state was evaluated by the Karnofsky Performance Scale and their quality of life according to Sintonen 15 D. Prevalence of at least mild depression before tumor operation was 30% for males and 38% for females. The mean depressive scores decreased significantly for up to one-year during follow-up for both males and females, but they remained notably high in all patients. Decreased functional status (KPS under 70) in the patients was significantly associated with high depressive scores at all measurement points. The decrease in the mean depressive scores was significant among patients with an anterior tumor and those with a pituitary adenoma. Five-year survival of the brain tumor patients was found to be mainly associated with the histology of the tumor. Survival time in months (SD) of the patients with high-grade (III–IV) gliomas was shown to be 22.5 (21.4), while it was 50.2 (19.9) for the patients with low-grade (I–II) gliomas, and 58.2 (9.4) for the rest of the patients. Depression among low-grade glioma patients was significantly associated with worse survival at five years follow-up. The level of anxiety was shown to be significantly higher among patients with a primary brain tumor in the right hemisphere compared to the anxiety scores among patients with left hemispheric tumors. A significant increase was found in the level of obsessionality over time in the female patients with a brain tumor in the left anterior location of the brain at three months after operation. The level of quality of life (QOL) was significantly worse among female brain tumor patients compared to males. Depressive females had significantly lower quality of life compared to that of non-depressive females up to one-year follow-up after surgical operation of the tumor. Depression, anxiety and obsessive-compulsive symptoms have to be recognized and be treated by psychotherapy and pharmacotherapy as soon as possible at every unit where brain tumor patients are followed and encountered.