Das kolorektale Karzinom (KRK) ist die zweithäufigste Krebserkrankung und die Metastasierung die häufigste Todesursache hierbei. Das neu identifizierte Gen MACC1 (metastasis associated in colon cancer 1) wurde als prognostischer Marker für die Metastasierung des KRK beschrieben. Im Zuge dieser Arbeit wurden die Exons 14-19 des Protoonkogens MET (met proto-oncogene (hepatocyte growth factor receptor)) und die kodierenden Exons von MACC1 in kolorektalen Tumoren sequenziert. Es waren in 60 Tumoren nur zwei MET Mutationen zu finden. In 154 kolorektalen Tumoren wurden die drei MACC1 single nucleotide polymorphisms (SNPs) rs47211888, rs975263 und rs3735615 identifiziert. Diese MACC1 SNPs veränderten nicht die MACC1 Expression in Tumoren oder KRK-Zelllinien. Sie waren nicht mit klinischen Daten von Patienten, nicht mit dem Gesamtüberleben oder dem metastasenfreien Überleben aller Patienten mit KRK assoziiert. Der MACC1 SNP rs975263 war signifikant mit einem kürzeren metastasenfreien Überleben in einer kleineren Gruppe von jüngeren Kolonkarzinom Patienten in frühen Stadien assoziiert. Zudem wurden mittels Microarray Analyse Targetgene von MACC1 identifiziert. MACC1 regulierte die Expression von S100P (S100 calcium binding protein P) und SPON2 (spondin 2, extracellular matrix protein) in den Zelllinien SW480 und SW620. Eine S100P oder SPON2 Überexpression förderte die Zellproliferation, Zellmigration und Zellinvasion. Intraspenal transplantierte Zellen mit hoher S100P oder SPON2 Expression führten im Gegensatz zu Kontrollzellen in Xenograft Modellen zur Bildung von Metastasen. Des Weiteren war die S100P oder SPON2 Expression in humanen metachron metastasierenden kolorektalen Tumoren höher als in nicht metastasierenden Tumoren. Patienten mit einer hohen S100P oder SPON2 Expression in ihren Tumoren hatten ein kürzeres metastasenfreies Überleben im Vergleich zu Patienten mit niedriger Expression. S100P und SPON2 könnten somit eine wichtige Rolle in der Metastasierung spielen. / Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the Western World, mainly due to metastasis. The gene MACC1 (metastasis associated in colon cancer 1) was described as a prognostic marker for CRC metastasis. In this study, we sequenced the exons 14-19 of the protooncogene MET (met proto-oncogene (hepatocyte growth factor receptor)) and the coding exons of MACC1 in colorectal tumors. We found two MET mutations in 60 tumors. In 154 tumors we identified the MACC1 single nucleotide polymorphisms (SNPs) rs47211888, rs975263 and rs3735615. These SNPs did neither modify the MACC1 expression in tumors nor in CRC cell lines. They were not associated with clinical parameters of the patients or with the overall survival and metastasis-free survival time of all CRC patients. Only in a subgroup, younger patients with colon cancer in early stages, the SNP rs975263 was significantly associated with a shorter metastasis-free survival time. Additionally, we identified new target genes of MACC1 by microarray analysis. MACC1 regulated the expression of S100P (S100 calcium binding protein P) and SPON2 (spondin 2, extracellular matrix protein) in the cell lines SW480 and SW620. Cell with a high S100P and SPON2 expression, intrasplenically transplanted into NOD/SCID mice, led to metastasis formation whereas transplanted control cells did not metastasize at all. The S100P and SPON2 expression was higher in colorectal tumors with metachronous metastasis than in non-metastasizing tumors. CRC patients with a high S100P or SPON2 expression in their primary tumors had a shorter metastasis-free survival time compared to patients with a low expression. Thus, S100P and SPON2 might play an important role in CRC metastasis.

Septinas constituem uma família de proteínas de ligação a GTP que foram inicialmente identificadas em levedura Saccharomyces cerevisiae, mas também estão presentes em outros eucariotos com exceção de plantas. Septinas são purificadas de leveduras, Drosophila e cérebros de mamíferos na forma de filamentos, porém o mecanismo através do qual acorre a formação destes filamentos ainda não é muito bem compreendido. Septinas são constituídas de três regiões principais: um N-terminal variável, um domínio central GTPase altamente conservado e um domínio coiled-coil C-terminal. O gene SEPT4 foi identificado por M. Tanaka e colaboradores a partir do cDNA de cérebro humano e apresentou duas distintas transcrições: Bradeiona ? e ?. Interessantemente, além de cérebro e coração, as proteínas Bradeiona Α e Β. são detectadas somente em câncer do cólon, reto, próstata e melanoma maligno. Neste trabalho, o gene da proteína Bradeiona Β foi subclonado em um vetor de expressão bacteriano, produzido em E. coli e purificado com sucesso. O espectro de dicroísmo circular (CD) mostrou o perfil característico de proteínas com hélices a na estrutura secundária. Resultados de cromatografia de exclusão molecular (SEC) e espalhamento dinâmico de luz (DLS) indicam que a septina Bradeina foi produzida na forma de um estável oligômero com características monodispersivas, que foi subseqüentemente cristalizado em PEG6000. A atividade GTPase da Bradeiona Β foi comprovada através da técnica de eletroforese capilar (CE), mostrando-se absolutamente dependente de íons Mg2+. Inibição da atividade GTPase foi verificada em altas concentrações de Mg2+ (maiores que 5 mM). Com a finalidade de caracterizar os domínios preditos da Bradeiona Β (Fragmento Conservado e domínio GTPase), essas regiões foram previamente definidas, expressas em E. cozi e purificadas com sucesso. Resultados de CD, SEC, espectroscopia de fluorescência e NMR-600MHz indicam que o FC foi produzido na forma de um estável monômero com pouca estrutura secundária regular. Resultados de DLS e CD indicam que a fusão 6xHis-DGTPase foi produzida na forma de um oligômero com a presença de hélices a na estrutura secundária. A fusão 6xHis-DGTPase mostrou-se instável a altas concentrações na ausência de imidazol. A atividade GTPase da fusão GST+DGTPase foi comprovada, similarmente a Bradeiona , através da técnica de CE. Novamente, verificou-se dependência de íons Mg2+ (para a atividade catalítica) e inibição em altas concentrações de Mg2+. A fusão GST+DGTPase também foi capaz de hidrolisar ATP. Espera-se que as informações relatadas neste estudo proporcionem um alicerce para estudos estruturais/funcionais futuros das proteínas Bradeiona Α e Βoutras septinas. / Septins form a class of eukaryoyic guanine nucleotide-binding proteins that were first identified in budding yeast. Septins purified from yeast, Drosophila and mammalian brain form filaments, however the mechanism by which the filaments assemble is unclear. Septins have a highly conserved structure, which includes a central GTP-binding domain, a variable N-terminal region, and most septins also contain a coiled coil domain at the Cterminus. Bradeion p is one of the splice variants of the human septin gene, SEPT4, recently isolated by expression screening of an adult human brain cDNA library. The Bradeion gene resides at 17q23, and has been shown to present specific expression in both human colorectal cancer, urologic cancers and malignant melanoma. In order to characterize the GTPase activity of Bradeion Β , the protein was successfully expressed in E. coli and purified. The recombinant protein was characterized by circular dichroism (CD), dynamic light scattering (DLS) and a novel non-radioactive enzyme assay, which utilizes capillary electrophoresis (EC) to monitor GTP hydrolysis. The CD spectrum exhibited the typical shape characteristic of the presence of helical elements of secondary structure and the DLS pattern was indicative of a monodisperse solution, which was readily crystallized in the presence of PEG6000. GTP hydrolysis was shown to be Mg2+ dependent within the low millimolar range but at 5 mM was inhibitory. In order to characterize the predicted domains of Bradeion Β, these defined regions were successfully expressed in E. cozi and purified. The CD spectrum of CF exhibited the shape typically found for non-regular structure. The results of fluorescence spectroscopy, gel filtration (SEC) and NMR-600MHz also corroborate with the CD results indicating an irregular structure. The fusion protein 6xHis-DGTPase exhibited a CD spectrum with the typical shape characteristic of the presence of helical elements but was show to be instable at high concentrations in the absence of imidazole. To characterize the GTPase activity of the fusion protein GST+DGTPase, the CE technique was used to monitor GTP hydrolysis. Analysis by CE showed that GST+DGTPase was functional, since both GTP and ATP hydrolysis was observed in a Mg2+ dependent manner. This work provides novel approaches, which should aid in the fbture study of the structure and fùnction of Bradeion Α e Β, others septins and related GTPases.

Le Cancer Colo Rectal (CCR) est la deuxième cause de mortalité par cancer dans le monde. Le risque de récidive après traitement curatif atteint 45% pour les stades 3. Une des hypothèses à l’heure actuelle pouvant expliciter le processus métastatique et les récidives est la présence en son sein de cellules « souches », pouvant « initier » le cancer. Notre réflexion s’inscrit dans la continuité des travaux réalisés au sein de notre Laboratoire, intitulés «Stratégies d’isolement et de caractérisation des cellules initiatrices de cancer colorectal», (Mélin et al, 2012). Dans une première partie, notre travail a porté sur l’analyse in vitro de la sensibilité des fractions enrichies en CIC aux différentes molécules de chimiothérapie les plus couramment utilisées en cancérologie colorectale.Puis, dans une deuxième partie, l’analyse des tumeurs obtenues après greffe a été faite ex ovo sur la Membrane Chorio-Allantoïdienne d’embryon de poulet (CAM), modèle facilement manipulable, peu onéreux et très rapide. Ce modèle étant naturellement immunodéprimé, il permet d’obtenir des informations concernant les phénomènes clés de la tumorigénèse et de la néoangiogénèse. Des analyses de la croissance tumorale, de l’histologie (prolifération, apoptose et vascularisation) et des analyses protéiques ont été menées en parallèle. De cette dernière étude, un marqueur particulier la E cadhérine, a été mis en évidence comme témoin indirect d’agressivité. En effet, au sein des tumeurs obtenues à partir de F1 HCT116, fraction himiosensible, l’expression de la E cadhérine est augmentée contrairement aux tumeurs obtenues à partir de F3 WiDr, fraction chimiorésistante, montrant une diminution d’expression de la E cadhérine.Ainsi, dans une troisième partie, sachant qu’un lien entre cellules initiant le cancer et E cadhérine a été mis en évidence, nous nous sommes focalisés sur son expression. Nous avons alors étudié son expression in vitro sur des cellules résistantes au 5 Fluorouracile. Puis, son expression a été étudiée ex vivo au sein de tissus et de sérums de patients opérés de cancer colorectal. / The ColoRectal Cancer (CCR) is the second leading cause of cancer mortality in the world. The risk of recurrence after curative treatment reaches 45% for stages 3. One of the hypotheses currently able to explain the metastatic process and the recurrences is the presence within it of "stem" cells, which can "initiate" the cancer. Our reflection is in line with the work carried out in our Laboratory, entitled "Strategies for the isolation and characterization of cells that initiate colorectal cancer" (Mélin et al, 2012).In the first part, our work focused on in vitro analysis of the sensitivity of fractions enriched in CIC to different chemotherapy molecules most commonly used in colorectal cancer (CRC). Then, in a second part, the analysis of the tumors obtained after transplantation was made ex ovo on Chicken Embryo Chorio-Allantoid Membrane (CAM), an easily manipulated model, inexpensive and very fast. This model provides information on the key phenomena of tumorigenesis and neoangiogenesis. Analyzes of tumor growth, histology (proliferation, apoptosis and vascularization) and protein analyzes were carried out in parallel. From this last study, a particular marker, E cadherin, was highlighted as an indirect link with witness of aggressiveness. Indeed, within the tumors obtained from F1 HCT116, the chemosensitive fraction, the expression of cadherin E was increased in contrast to the tumors obtained from F3 WiDr, chemoresistant fraction, showing a decrease in expression of E cadherin.Thus, in a third part, knowing that a link between cells initiating cancer and E cadherin was highlighted, we focused on its expression. We first studied its expression in vitro on 5 Fluorouracilresistant cells. Its expression was then studied ex vivo in tissues and sera of operated patients of CRC.

La chronothérapie des cancers administre les médicaments anticancéreux à des moments précis de la journée afin d’en optimiser la tolérance et l’efficacité. Cependant le système circadien qui règle sur 24 h la prolifération et le métabolisme cellulaires peut être altéré par un décalage horaire chronique, la mutation d’un gène de l’horloge, ou un traitement, favorisant ainsi la survenue de pathologies métaboliques, comportementales ou malignes. La disparité des profils circadiens de température corporelle des patients cancéreux ainsi que leurs modifications sous chimiothérapie fournit les bases d’une personnalisation de la chronothérapeutique. La capacité d’un cycle thermique à entraîner sur 24 h l’horloge circadienne de cellules d’hépatocarcinome en culture indique que ce biomarqueur est aussi un effecteur de la synchronisation des cellules cancéreuses, et constitue un repère circadien pour la chronothérapeutique in vitro et in vivo. / Chronotherapy delivers anticancer drugs at specific times of the day to optimize tolerability and efficacy. However, the circadian system that controls cell proliferation and metabolism over 24 h, can be altered by a chronic jet lag, a clock gene mutation, or a xeniobiotic treatment, thus favoring the occurrence of metabolic, behavioral or malignancies. The disparity of circadian body temperature patterns of cancer patients as well as its disruption during the treatment provides a clincher for chronotherapy personalization. The ability of a thermal cycle to drive the circadian clock in cultured hepatocarcinoma cells of 24 h indicates that this biomarker is also an effector of the synchronization of cancer cells, as well as a marker for the circadian in vitro and in vivo chronotherapeutic.

O objetivo do estudo foi avaliar o efeito da escuta terapêutica na ansiedade pré-operatória e nos medos relacionados à cirurgia em pacientes hospitalizados para tratamento cirúrgico de câncer colorretal. Tratou-se de um ensaio clínico aleatorizado realizado em um hospital geral. Os participantes foram aleatorizados em dois grupos: experimental (GE) (n=25) e controle (GC) (n=25). No GE era realizada a escuta terapêutica, de modo que os pacientes tinham 30 minutos para falar com a pesquisadora sobre sua experiência com a hospitalização para o tratamento da doença. No GC os pacientes eram informados que teriam alguns dados coletados, em seguida a pesquisadora se ausentaria por 30 minutos e que, após este intervalo, a mesma retornaria para a conclusão da pesquisa. As variáveis dependentes (alfa-amilase, cortisol, frequência de pulso, frequência respiratória, pressão arterial, ansiedade estado e medos relacionados à cirurgia) foram coletadas antes e após a realização da intervenção no GE e antes e após o intervalo supracitado no GC. Na comparação das variáveis dependentes no GE em relação ao GC no momento pós-intervenção, não foram identificadas diferenças significativas para nenhuma das variáveis estudadas. Na comparação das variáveis no GE e GC nos momentos pré e pós-intervenção, constatou-se diferenças significativas apenas entre os momentos no GC para as variáveis cortisol (Z=-2,023; p=0,043), frequência de pulso (FP) (Z=-2,121; p=0,034) e medos relacionados à cirurgia (Z=-2,171; p=0,030), com redução dos valores destas variáveis. Na relação entre as variáveis estudadas obteve-se significância entre a ansiedade estado e as variáveis cortisol, idade, escolaridade e religião; os medos relacionados à cirurgia foram relacionados ao sexo, tempo de confirmação do diagnóstico de câncer colorretal, nível de escolaridade, cortisol, estado civil e à pressão arterial diastólica. Apenas a variável renda familiar mensal apresentou diferença significativa em suas distribuições no GE e no GC, no entanto, esta não teve influência sobre as variáveis dependentes. A maioria dos pacientes referiu satisfação quanto ao desenvolvimento da intervenção. Conclui-se que, no momento pré-operatório, nas condições investigadas neste estudo, os dados evidenciaram que a mensuração das variáveis imediatamente após a intervenção de escuta terapêutica de 30 minutos não evidenciou a redução esperada dos valores das variáveis estudadas; estudos com medidas dessas variáveis após um tempo para o paciente processar os efeitos da intervenção poderão confirmar se a mensuração imediata ou uma hora após a intervenção, por exemplo, conduziria a resultados distintos. Contudo, houve redução dos valores de cortisol, frequência de pulso e medos relacionados à cirurgia nos participantes do GC, o que pode ser atribuído ao contato da pesquisadora com tais participantes durante os momentos de coleta de dados / The aim of the study was to evaluate the effect of therapeutic listening on preoperative anxiety and fears related to surgery in patients hospitalized for surgical treatment of colorectal cancer. This is a randomized clinical trial conducted in a general hospital. Participants were randomized in two groups: experimental (n=25) and control (n=25). In the experimental group was performed therapeutic listening. The patients had thirty minutes to talk to the researcher about their experience with hospitalization for the treatment of disease. In the control group patients were informed that they would have some data collected, then the researcher would be gone for thirty minutes and after this range, she would return to complete the research. The dependent variables (alpha-amylase, cortisol, pulse rate, respiratory rate, blood pressure, anxiety state and fears related to surgery) were collected before and after the intervention in experimental group and before and after the break in control group. In comparing the dependent variables in the experimental group compared to the control group in the post-intervention time, no significant differences were identified for any of the variables studied. In comparing the variables in the experimental group and the control group in the pre and post-intervention, there was significant differences only between times in the control group for cortisol (Z=-2,023; p=0,043), pulse rate (Z= -2,121; p=0,034) and fears related to surgery (Z= -2,171; p=0,030), with reduced of these variables. In the relationship between the studied variables, there was a significant difference between state anxiety and the variables cortisol, age, education level and religion; the fears related to the surgery were related to sex, time of confirmation of the diagnosis of colorectal cancer, education level, cortisol, marital status and diastolic blood pressure. Only the variable monthly income showed a significant difference in their distributions in the experimental group and the control group, however, this had no influence on the dependent variables. Most patients report satisfaction with the development of the intervention. It was concluded that, in the preoperative period, under the conditions investigated in this study, the data showed that the measurement of the variables immediately after the intervention of 30 minutes of therapeutic listening did not show the expected reduction of the values of the studied variables; Studies with measures of these variables after a time for the patient to process the effects of the intervention could confirm whether the immediate measurement or one hour after the intervention, for example, would lead to different results. However, there was reduction of cortisol levels, pulse rate and fears related to surgery in the control group, which can be attributed to contact the researcher with these participants during times of data collection

Inscription de la thèse. La présente thèse a fait l’objet d’une Convention Industrielle de Formation par la Recherche (CIFRE) entre le laboratoire de Psychologie EA-4139 Santé et Qualité de Vie de Bordeaux et l’Association pour le Dépistage du Cancer Colorectal (ADECA Alsace) à Colmar.Objectifs. L’objectif principal de la thèse était d’améliorer la participation des 50-75 ans au programme de dépistage organisé du cancer colorectal en Alsace, cela dans le respect de leur autonomie de décision. La thèse était aussi l’occasion de mieux cerner les facteurs d’adhérence aux recommandations de santé. Plan de la thèse. La thèse se compose en deux grandes parties. Dans une première partie, théorique, nous nous sommes interrogés sur la place de la prévention dans nos sociétés hypermodernes (Aubert, 2006) tout en proposant une lecture intégrative de l’adhérence à travers le concept de motivation (Carré et Fenouillet, 2009). Le modèle articule en particulier les apports de l’autorégulation (Hall et Fong, 2007), de l’autodétermination (Ryan et Deci, 2000) et de la dissonance (Festinger, 1957). Il adresse aux professionnels de santé un certain nombre de préconisations destinées à les aider à optimiser leur communication de prévention. Dans une seconde partie, empirique, nous présentons les études randomisées contrôlées mettant à l’épreuve des faits les éléments du modèle articulateur. Deux stratégies motivationnelles ont été testées. Toutes deux greffées sur le dépistage organisé du cancer colorectal en Alsace. La première étude (N=48 413) a fait l’objet d’un article publié dans la revue médicale Gastroenterology et de deux communications, dont une internationale à la Disease Digestive Week de San Diego. Elle évalue l’apport d’un conseil téléphonique ajusté guidée par une approche transthéorique de la motivation (Weinstein, 1988), en particulier l’entretien motivationnel (Miller et Rollnick, 2006). La deuxième étude randomisée contrôlée (N=22397) est une intervention motivationnelle postale axée sur l’autonomie et l’implication. Elle s’inspire du choix informé (Gøtzsche et al., 2008) et de la communication engageante (Girandola et Joule, 2008). Principaux résultats. Les résultats démontrent dans la première étude qu’un conseil téléphonique personnalisé décuple le retour des exclusions médicales (19,2%>1,8%) χ2 (1, N=26425) = 2,603E3 ; p=.000 et permet de doubler, voire tripler la participation par rapport au courrier (30,4%>9,2%) χ2 (1, N=22535) = 1,140E3 ; p=.000 quel que soit le type de conseil χ2 (1, N=2 182) =1,195 ; p=.274. La procédure témoigne en revanche d’un réel écueil de faisabilité par rapport au courrier (3,8%<9,2%) χ2 (1, N=46773) = 5,781E2 ; p.000. La deuxième étude montre un bénéfice du choix informé sur la satisfaction du sujet eu égard au processus d’information F (4, N=63) = 8,570 ; p=.000 mais aucun effet du type de courrier sur la participation en phase de première invitation χ2 (4, N=11470) = 3,012 ; p=.556 comme en relance χ2 (4, N=10610) = 4,352 ; p=.360. Discussion. Les résultats sont discutés et interprétés à l’appui des considérations théoriques du modèle articulateur. Des préconisations pour les professionnels et pour des recherches futures sont soumises en fin de thèse. / Context of the thesis: The present thesis is part of an Industrial Convention of Formation by Research (CIFRE) between the laboratory of Psychology EA-4139 Santé et Qualité de Vie of Bordeaux and the Association pour le Dépistage du Cancer Colorectal en Alsace (ADECA Alsace) in Colmar.Aims of the study: The main objective of the thesis was to improve the involvement of the 50-75 years old segment of the population of Alsace in the organized colorectal cancer screening, whilst respecting their personal decision. The thesis was also the opportunity to get a better understanding of the adherence factors to health recommendations. Structure of the thesis: The thesis is made up of two main parts. In a first theoretical part, we looked into the place of the prevention in our hypermodern societies (Aubert, 2006) while proposing an integrative reading of adherence through the concept of motivation (Carré & Fenouillet, 2009). The model underlines especially the contribution of self-regulation (Hall & Fong, 2007), self-determination (Ryan & Deci, 2000) and dissonance (Festinger, 1957). It provides health professionals with a wide range of recommendations to help them optimize their communication on prevention. In a second empirical part, we have put to the test the components of the articulator model by the randomized controlled studies and, in this way, tested two motivational strategies both related to organized colorectal cancer screening program in Alsace. The first study (N=48 413) resulted in an article published in the Gastroenterology medical review and two conferences, one of which in the international Disease Digestive Week in San Diego. It evaluates the contribution of an adjusted phone counselling guided by a transtheoric approach of motivation (Weinstein, 1988), amongst other things motivational interviewing (Miller & Rollnick, 2006). The second randomized controlled study (N=22397) is a postal motivational intervention focused on self-reliance and involvement inspired by the informed choice (Gøtzsche et al., 2008) and the engaging communication (Girandola & Joule, 2008). Main outcomes: The results of the first study demonstrate that an individualized phone counselling enables to get a tenfold increase in the return of medical exclusions (19,2%>1,8%) χ2 (1, N=26425) = 2,603E3 ; p=.000 and to double or even triple the participation compared to mail (30,4%> 9,2%) χ2 (1, N=22535) = 1,140E3; p=.000 whatever the type of advice χ2 (1, N=2 182) =1,195; p=.274. However, the procedure encounters barriers with regard to its feasibility compared to mail (3,8%< 9,2%) χ2 (1, N=46773) = 5,781E2; p.000. The second study shows a benefit of the informed choice on the satisfaction of the subject related to the information process F (4, N=63) = 8,570 ; p=.000 but no effects of the type of mail on the participation in the course of the phase of first invitation χ2 (4, N=11470) = 3,012 ; p=.556 and even in the relaunching process χ2 (4, N=10610) = 4,352 ; p=.360. Discussion: The results are discussed and interpreted with the support of the theoretical consideration of the articulator model. Recommendations for health professionals and further research projects are submitted at the end of the thesis.

Introdução: O câncer de localização colorretal é o terceiro tipo de câncer mais comumente diagnosticado no mundo. As taxas de incidências do câncer colorretal não são homogêneas, apresentando diferenças entre os países. Não há estudos brasileiros que investiguem a variação geográfica da incidência de câncer colorretal conjuntamente com indicadores socioeconômicos. Esta avaliação pode revelar diferenças locais importantes na ocorrência da doença. Objetivos: Descrever as taxas de incidência e de mortalidade do câncer colorretal no Município de São Paulo, segundo sexo e faixa etária, no período de 1997 a 2009 e realizar análise da distribuição espacial segundo distrito dos casos de câncer colorretal diagnosticados em residentes no Município de São Paulo entre 1997 e 2009. Material e Métodos: Foram analisados os novos casos de câncer colorretal diagnosticados em residentes no Município de São Paulo de 1997 a 2009. Estes dados foram fornecidos pelo Registro de Câncer de Base Populacional de São Paulo (RCBP-SP). A análise dos dados foi realizada em duas etapas: na primeira, com cárater exploratório/descritivo, os dados analíticos foram utilizados para descrever a incidência e mortalidade por câncer colorretal no período pesquisado. Na segunda etapa, os casos de câncer colorretal foram geocodificados, agrupados por distrito administrativo e estudados segundo a metodologia de análise para dados de área. Toda análise foi implementada no software R. Resultados: Com 7,7 por cento e 7,3 por cento dos casos respectivamente em homens e mulheres, câncer colorretal foi o segundo tipo de câncer mais frequente, sendo a quarta (9,0 por cento dos óbitos) e a segunda (11,0 por cento dos óbitos) causa de morte respectivamente em homens e mulheres. Do total de casos incidentes (39.250), 47,50 por cento são do sexo masculino e 52,50 por cento do sexo feminino. Destes, 4.784 (37,7 por cento ) evoluíram a óbito, sendo 48,1 por cento no sexo masculino e 51,9 por cento no sexo feminino. As taxas específicas por sexo e faixa etária de incidência aumentam fortemente com a idade, na faixa etária de 80 ou mais anos chega a 377,9 e 282,9 (por 100 mil hab.) para o sexo masculino e feminino respectivamente, sendo relativamente próximas em ambos os sexos até a idade de 49 anos e maiores para homens nas faixas etárias subsequentes. As taxas específicas por sexo e faixa etária de mortalidade, apresentam comportamento análogo, aumentam fortemente com a idade, na faixa etária de 80 ou mais anos chega a 206,9 e 159,9 (por 100 mil hab.) para o sexo masculino e feminino respectivamente. A taxa anual de incidência ajustada pela população de SEGI (1960) e modificada por DOLL et al. (1966) apresenta-se em torno de 30,0 (por 100 mil hab.) nos três primeiros anos observados (1997-1999), chega a 19,0 (por 100 mil hab.) em 2002, volta a crescer nos anos seguintes (2003-2005), chegando a 31,7 (por 100 mil hab.) e matem-se estável de 2007 a 2009. A taxa anual de mortalidade de câncer colorretal ajustada pela população crescente até 2004, chegando a 15,7 (por 100 mil hab.) e decrescem nos anos seguintes, chegando a aproximadamente 3,6 mortes por 100 mil habitantes em 2009. A média anual da taxa bruta de incidência e os indicadores socioeconômicos apresentam dependência forte dependência espacial, sendo o menor Índice I de Moran observado foi para o índice de exclusão/inclusão dos anos potenciais de vida perdidos (IEX apvp = 0,29), os demais são acima de 0,6. Os indicadores apresentam forte correlação linear com a média anual da taxa bruta de incidência. Conclusões: As distribuições da incidência e da mortalidade apresentam padrões semelhantes ao identificado mundialmente. O Município de São Paulo tem taxas equivalentes às encontradas nas regiões em transição econômica. Foi identificada forte dependência espacial na distribuição da incidência de câncer colorretal no Município de São Paulo, com a formação de clusters nas áreas centrais e periféricas. As maiores taxas são encontradas nas áreas centrais e nas periferias. A distribuição espacial da incidência de câncer colorretal apresenta forte associação com a distribuição dos indicadores de status socioeconômico no Município de São Paulo, em particular apresenta associação positiva com indicadores de renda e escolaridade. / Introduction: Colorectal cancer is the third most common diagnosed cancer worldwide. Colorectal cancer incidence rates are not homogeneous, with differences between countries. No Brazilian studies investigated the geographical variation of colorectal cancer incidence with socioeconomic indicators. This study may reveal important local differences in the occurrence of the disease. Objectives: To describe colorectal cancer incidence and mortality in São Paulo, by sex and age using 1997-2009 data and perform the spatial distribution analysis according to district colorectal cancer cases diagnosed in residents at Municipality of São Paulo between 1997 and 2009. Methods: Colorectal cancer cases diagnosed from 1997 to 2009 in São Paulo residents were analyzed. These data were provided by Population Based Cancer Registry of São Paulo (RCBP-SP). Data analysis was performed in two stages. First, analytical data were used to describe the incidence and mortality from colorectal cancer. Second, colorectal cancer cases were geocoded, grouped by administrative district and studied according data area analysis methodology. All analysis was implemented in software R. Results: 7.7 per cent and 7.3 per cent of observed cases was respectively in men and women, colorectal cancer was the second most common cancer, the fourth (9.0 per cent ) cause of death in men and the second (11.0 per cent ) cause in women. It was diagnosed 39,250 colorectal cancer new cases, 47.50 per cent in men and 52.50 per cent in women. And 4,784 (37.7 per cent ) died, with 48.1 per cent in male and 51.9 per cent in female. The specific incidence rates strongly increase with age, at the 80 years or more age reaches 377.9 and 282.9 (per 100,000 inhabitants) for male and female respectively. The mortality specific rates, have similar behavior, strongly increase with age and at the 80 years or more age reaches 206.9 and 159.9 (per 100,000 inhabitants), for males and female respectively. The annual age adjusted incidence rate was around 30.0 (per 100,000 inhab.) in the first observed years (1997-1999), arrives to 19.0 (per 100,000 inhab.) in 2002, grow back reaching 31.7 (per 100,000 inhab.) and kill stable from 2007 to 2009. The annual age colorectal cancer mortality rate grow reaching 15.7 (per 100,000 inhab.) and decrease in the following years, reaching approximately 3.6 deaths per 100,000 inhabitants in 2009. The average annual the crude incidence rate and the socio-economic indicators show strong spatial dependence, the lowest Moran´s I Index was observed for the exclusion/inclusion potential years of life lost index (IEX apvp = 0.29). The indicators show strong linear correlation with the average annual crude incidence rate. Conclusions: Distributions of incidence and mortality have similar worldwide patterns. The Municipality of São Paulo has equivalent rates founded in regions in economic transition. It was identified strong spatial dependence in the distribution of the incidence of colorectal cancer, with the formation of clusters in the central and peripheral areas of Municipality of São Paulo. The highest rates were found in the central areas and lowest were found in the suburbs. The spatial distribution of colorectal cancer incidence has a strong association with the socioeconomic status indicators distribution in Municipality of São Paulo. It was identified positive association between colorectal cancer incidence with income and education indicators.

L’acide docosahexaénoïque (DHA) est un acide gras polyinsaturé oméga-3 avec des propriétés anti-inflammatoires et anti-tumorales. L’effet du DHA dans le cadre du cancer colorectal pourrait être la conséquence d'une action anti-proliférative directe sur les cellules cancéreuses et de sa capacité à réduire l’inflammation propice au développement de la tumeur. Le Tumor Necrosis Factor-alpha (TNFa) est une cytokine pro-inflammatoire et présente des effets paradoxaux. En fonction du contexte cellulaire, le TNFa activera une voie de signalisation dépendante de la kinase RIP1 engageant la cellule cancéreuse vers la prolifération ou la mort cellulaire. Notre objectif fut d'évaluer le rôle du TNFa dans l'effet anti-prolifératif du DHA sur des cellules cancéreuses coliques et de préciser les mécanismes moléculaires régulant l'expression de cette cytokine. Le DHA induit l'expression de TNFa et sa sécrétion par les cellules cancéreuses. Nous avons montré que des anticorps neutralisant l'action autocrine du TNFa sur les cellules cancéreuses prévenait l'effet pro-apoptotique du DHA et abolissait l'effet anti-cancéreux observé dans des souris nude avec tumeurs HCT-116 sous régime DHA. L’induction de l'expression de TNFa par le DHA prend son origine à un niveau post-transcriptionnel par la répression du microARN miR-21 perdant sa capacité à dégrader l'ARNm TNFa. Le DHA par l'activation des kinases AMPKa et RIP1 déclenche la translocation nucléaire du facteur de transcription FOXO3a se fixant sur le promoteur miR-21 et diminuant l’expression de ce microARN. Nos travaux mettent en évidence un nouveau mécanisme moléculaire soutenant l'action anti-tumorale du DHA. / Docosahexaenoic acid (DHA) is an omega-3 polyunsaturated fatty acid with anti-inflammatory and anti-tumoral properties. The anti-tumor effect of DHA in colorectal cancer might be attributed to direct anti-proliferative action on cancer cells and to its ability to reduce inflammatory status involved in tumor growth. Tumor Necrosis Factor-alpha (TNFa) is an inflammatory cytokine with paradoxical effect in cancer biology. According to the cellular context, TNFa activates RIP1 kinase dependent signaling pathway leading to proliferation or cell death. Our aim was to evaluate the role of TNFa in anti-proliferative effect of DHA in colon cancer cells and to precise the molecular mechanisms regulating TNFa expression.DHA treatment increased TNFa expression and secretion by cancer cells. We have shown that neutralization of autocrine TNFa action prevented the pro-apoptotic effect of DHA colon cancer cells and abolished anti-cancer effect in tumor HCT-116 bearing nude mice fed a DHA-enriched diet. Induction of TNFa expression by DHA occured at post-transcriptional level through microRNA miR-21 repression reducing its ability to induce TNFa mRNA degradation. DHA activates AMPKa and RIP1 kinases triggering nuclear translocation of the transcription factor Foxo3a which bound to miR-21 promoter and repressed the microRNA expression. Our works highlight a new molecular mechanism supporting the anti-cancer action of DHA.

Die Leber nimmt als Hauptzielorgan des metastasierenden Kolorektalkarzinoms (CRC) eine exponierte Rolle ein; bei mehr als 50 % der betroffenen Patienten werden im Laufe ihrer Tumorerkrankung Lebermetastasen diagnostiziert, die trotz multimodaler Therapiekonzepte immer noch den fatalen Kranksheitsverlauf bestimmen. Zwecks Entwicklung neuer Therapiestrategien ist ein fundiertes Verständnis der Entstehungsmechanismen und des Genexpressionsprofils der Metastasen erforderlich. Makrophagen wird in diesem Kontext ein großer Einfluss auf die Tumorentstehung und -progression zugeschrieben, weshalb von einer klinischen Relevanz der Makrophagenpolarisation auszugehen ist. Im Rahmen der vorliegenden Arbeit wurde mit der CRC Zelllinie CMT-93 ein syngenes, orthotopes Lebermetastasenmodell in C57BL/6N Mäusen etabliert. Basierend auf RNA Sequenzierungsdaten und bioinformatischen Analysemethoden wurde eine Entwicklung und fein abgestimmte Adaptation der CMT-93 Zellen im Zuge ihrer Propagation im hepatischen Milieu nachgewiesen. Diese resultierte in divergierenden Genexpressionsprofilen zwischen Zelllinie und Metastasen. Von insgesamt 3329 differentiell exprimierten Genen wurden mittels einer Selektionsliste 32 signifikant exprimierte Gene identifiziert. Insbesondere Matrix-Metalloproteasen (MMP-2, -7, -9), Chemokinrezeptoren (CXCR2, CXCR4), Zelladhäsionsgene (ITGA6, ITGB3) sowie Wif1 als Feinregulator des kanonischen Wnt Signalweges nehmen eine bedeutende Rolle ein und tragen zum Invasions- und Metastasierungsprofil von CMT-93 Zellen unter dem Einfluss des Lebermilieus bei. Invasive und aggressive Eigenschaften der CMT-93 induzierten Lebermetastasen wurden insbesondere im frühen und späten Metastasierungsstadium nachgewiesen. Hier wurde u.a. die Expression der EMT Marker Vimentin, MMP-7 und CD44, Ki-67, NFkb1 und Stat3 untersucht. Eine Gene Ontology Analyse und RNA Sequenzierungsdaten verschiedener Leberareale zeigten, dass die vielschichtige Interaktion zwischen CMT-93 Zellen und der hepatischen Mikroumgebung u.a. durch immunregulatorische Prozesse gesteuert wird, die in veränderten Genexpressionsprofilen zwischen Zelllinie, Metastase und tumorumgebendem Gewebe resultiert. In Lebermetastasen konnte eine Mischpopulation aus M1 und M2 Makrophagen nachgewiesen werden, die einen tendenziell M2 geprägten Charakter aufweisen. Dieser ist höchstwahrscheinlich an einer Stimulation der invasiven Eigenschaften der Tumorzellen beteiligt. Durch qRT-PCR und immunhistochemische Analysen konnten Hinweise gesammelt werden, dass nicht explizit die Quantität oder spezifische Lokalisation von Makrophagenphänotypen, sondern mutmaßlich eher das Zytokinprofil des Tumors und der Mikroumgebung und damit einhergehend der Polarisationsstatus der Makrophagen zum Metastasierungserfolg beiträgt. Eine Inkubation von CMT-93 Zellen mit konditionierten Medien der verschiedenen Makrophagenphänotypen bestätigte die Rolle sezernierter Faktoren. Eine indirekte Interaktion zwischen Tumorzelle und M2 Makrophagen reicht offensichtlich aus, um tumorstimulierende Eigenschaften, Aggressivität und Proliferationsfähigkeit der Zelllinie zu beeinflussen. Somit wurden Gene und Signalwege identifiziert, die relevant sind für eine erfolgreiche Induktion und Progression von Lebermetastasen infolge der Implantation von CRC-Zellen. Eine Adaptation des Genexpressionsprofils der CMT-93 Zelllinie im Zuge der Leberkolonisation konnte nachgewiesen werden. Das molekulare Profil bzw. die Gensignatur der Metastasen korrelierte dabei in hohem Maße mit der Migrations- und Invasionsfähigkeit der Tumorzellen. Insbesondere die Anwendung bioinformatischer Methoden erwies sich dabei als nützliches Werkzeug zur Analyse von großen Datensätzen, die mittels RNA Sequenzierung generiert wurden. Diese werden nun zur Formulierung prädiktiver Modelle der Metastasierungsvorgänge genutzt, um deregulierte Gene und damit einhergehend die Aggressivität von Tumorzellen gezielt identifizieren und konsekutiv beeinflussen zu können. Eine Analyse der Makrophagenphänotypen und ihrer Interaktion mit Tumorzellen verdeutlichte den durchaus tumorstimulierend geprägten Charakter der CMT-93 induzierten Lebermetastasen und den Einfluss des Zytokinprofils auf die Tumorprogression. Eine weiterführende Untersuchung der Makrophagenpolarisation bedarf jedoch eine rigorose Charakterisierung der Phänotypen anhand weiterer spezifischer Marker. Die subtile Analyse wird Rückschlüsse der organspezifischen Einflüsse des Zytokinprofils auf die kolorektale Karzinogenese erlauben.

Submitted by Michele Mologni (mologni@unoeste.br) on 2019-01-29T18:40:59Z No. of bitstreams: 1 Sandra Cristina Genaro.pdf: 1632390 bytes, checksum: 89f6ec224705d20a5ce967976b70a4c3 (MD5) / Made available in DSpace on 2019-01-29T18:40:59Z (GMT). No. of bitstreams: 1 Sandra Cristina Genaro.pdf: 1632390 bytes, checksum: 89f6ec224705d20a5ce967976b70a4c3 (MD5) Previous issue date: 2018-11-22 / Among the existing types, colorectal cancer (CRC) affects approximately one million people per year, considered the second most common cause of death among women and the third most prevalent in men. Risk factors are genetic syndromes; inflammatory bowel diseases; family history; sedentary lifestyle; obesity, low fiber, high saturated fats, smoked foods or built-in food, excess red meat (> 300g/week), preparation mode in high temperatures and on the ember; medications; smoking, and excessive alcohol. These factors alter the intestinal microbiome which is colonized by pathogenic bacteria capable of provoking a local inflammatory response that, in chronic cases, activates carcinogenic components. Probiotics have increasingly attracted the attention of researchers in order to understand their action in the intestinal microbiota, aiding in the prevention and treatment of colorectal cancer. The objective of this study was to evaluate the effect of a probiotic In aggressiveness of the chemically induced colorectal tumor in rats. Twenty-five male Fisher 344 rats, 250 g, receiving ration and water ad libitum, were randomly divided into 5 groups (5 rats/group): GControl, without treatment; GTumor, tumor induction; GTumor + 5FU, tumor induction, 5-fluorouracil applied; GTumor + prob, tumor induction, supplemented with probiotic; GTumor + 5-FU + prob, tumor induction, applied 5-fluorouracil, supplemented with probiotic. For tumor induction, the animals received four intraperitoneal injections of the carcinogen 1.2-dimethylhidrazine (DMH) at the dose of 20 mg/kg body weight, being two applications per week, for four consecutive week. A 15-day interval was given and DMH applications were repeated for another four week. After 5 weeks of the last dose of the carcinogen, the treatment was initiated for ten consecutive weeks, applying a weekly dose of 15 mg/kg body weight of 5-fluorouracil, Intraperitoneal route and commercial probiotic containing Lactobacillus and Bifidobacterium at the dose of 1x109 UFC, administered by gavage, daily. Datas were analyzed by the analysis of variance One Way and the averages compared by the test of Dunnett. Used Software Statistical GraphPad Prism. The histopathologic analyses evaluated by the Chi-square ratio test. It was considered type-I error of 5% as statistically significant. Compared to the GTumor, the GTumor + prob (P < 0,0373) and GTumor + 5-FU + prob (P < 0,0003) showed attenuating effect on the aggressiveness of the colorectal tumor, with a reduction in the count of Aberrant Crypts Foci; and lower percentage of malignant neoplastic lesions in the GTumor + prob (40% of low-grade tubular adenoma, 40% of carcinoma in situ, 20% of low-grade adenocarcinoma) and GTumor + 5-FU + prob (40% of low-grade tubular adenoma and 60% of carcinoma in situ). The suplementation with probiotic has the potential to decrease the formation of aberrant crypts and mitigate the progression of tumor malignancy, potentializing the antitumor effect of 5-fluorouracil chemotherapy in the colic segments. / O câncer colorretal (CCR) acomete aproximadamente um milhão de pessoas por ano, considerado a segunda causa de morte mais comum entre mulheres e a terceira mais prevalente em homens. Os fatores de risco incluem as síndromes genéticas; doenças inflamatórias intestinais; história familiar; sedentarismo; obesidade, alimentação pobre em fibras, rica em gorduras saturadas, alimentos defumados ou embutidos, carne vermelha em excesso (>300g/sem), modo de preparação em altas temperaturas e na brasa; medicamentos; tabagismo e bebida alcoólica em excesso. Esses fatores levam a alteração da microbiota intestinal a qual é colonizada por bactérias patogênicas capazes de provocar uma resposta inflamatória local que, em casos crônicos, ativam componentes cancerígenos. Os probióticos têm atraído cada vez mais a atenção de pesquisadores com o intuito de compreender a sua ação na microbiota intestinal, auxiliando na prevenção e tratamento do câncer colorretal. O objetivo desse estudo foi avaliar o efeito de um probiótico na agressividade do tumor colorretal induzido quimicamente em ratos. Vinte e cinco ratos machos Fisher 344, 250 g, recebendo ração e água ad libitum, foram divididos aleatoriamente em 5 grupos (5 ratos/grupo): GControle, sem tratamento; GTumor, indução do tumor; GTumor+5FU, indução do tumor, aplicado 5-Fluorouracil; GTumor+Prob, indução do tumor, suplementado com probiótico; GTumor+5-FU+Prob, indução do tumor, aplicado 5-Fluorouracil, suplementado com probiótico. Para indução do tumor colorretal, os animais receberam quatro injeções intraperitoneais do carcinógeno 1,2-dimetilhidrazina (DMH) na dose de 20 mg/kg de peso corporal, sendo duas aplicações por semana, durante quatro semanas consecutivas. Deu-se um intervalo de 15 dias e as aplicações de DMH foram repetidas por mais quatro semanas. Após 5 semanas da última dose do carcinógeno, iniciou-se o tratamento por dez semanas consecutivas, com 5-Fluorouracil: uma dose de 15 mg/kg por semana, via intraperitoneal e probiótico comercial: 1x109 UFC, diariamente, por gavagem. Os dados foram analisados pela Análise de Variância One Way e as médias comparadas pelo teste de Dunnett. Utilizado software estatístico GraphPad Prism. As análises histopatológicas avaliadas pelo teste de proporção Qui-quadrado. Foi considerado erro tipo-I de 5% como estatisticamente significante. Comparados com o GTumor, o GTumor+Prob (p<0,0373) e GTumor+5-FU+Prob (p<0,0003) exibiram efeito atenuante na agressividade do tumor colorretal obervando redução na contagem de Focos de Criptas Aberrantes; e menor porcentagem de lesões neoplásicas malignas no GTumor+Prob (40% de adenoma tubular de baixo grau, 40% de carcinoma in situ, 20% de adenocarcinoma de baixo grau) e GTumor+5-FU+Prob (40% de adenoma tubular de baixo grau e 60% de carcinoma in situ). Concluimos que a suplementação com probiótico tem potencial para diminuir a formação de criptas aberrantes e amenizar a progressão da malignidade do tumor, potencializando o efeito antitumoral da quimioterapia com 5-Fluorouracil nos segmentos cólicos.