O objetivo deste estudo foi investigar as relações entre os parâmetros de halitose e sialometria em pacientes submetidos à radioterapia de cabeça e pescoço, avaliando a presença de saburra, classificada em termos de peso seco, fluxo salivar, teste BANA e halimetria oral e comparar os resultados entre os grupos. Foram examinados 48 pacientes divididos em três grupos. O grupo 1 (\"saúde\") foi constituído de pacientes que haviam terminado o tratamento na Faculdade de Odontologia de Bauru (FOB), sem queixa de halitose e com íntegra saúde oral; o grupo 2 (\"N. O.\") por indivíduos com necessidades odontológicas e o grupo 3 (\"radio\") pacientes que haviam sido submetidos à radioterapia na região de cabeça e pescoço. Foram avaliadas as concentrações bucal de Compostos Sulfurados Voláteis (CSVs) através do monitor portátil de sulfetos HalimeterTM, os valores relacionados à taxa de fluxo salivar em repouso e estimulado, teste BANA com material proveniente da saburra lingual e peso seco da mesma. Os resultados foram analisados utilizando o teste de análise de variância que quantificaram o hálito antes e após a remoção da saburra. As relações entre halimetria bucal, peso seco da saburra, fluxo salivar com e sem estímulo, e a relação entre fluxo salivar em repouso e estimulado foram feitas através do teste de correlação de Pearson. Para verificar correlações entre halimetria bucal inicial e teste BANATM foi utilizado o teste \"t\" de Student. Os resultados mostraram que houve diferença estatisticamente significante entre a halimetria oral nos grupo \"saúde\" e \"radio\" e, nos grupos \"saúde\" e \"N.O.\" Pode-se observar também que houve relação entre a presença de saburra e os níveis de CSVs. Em relação ao fluxo salivar, foi observada uma redução estatisticamente significante entre o grupo \"radio\" e os outros dois. Nas condições dessa pesquisa, foi constatado que a halitose pode ser considerada um efeito adverso da radioterapia, vinculada à hipossalivação e saúde oral deficiente. / The aim of this study was to verify the halitosis parameters in patients who had received head and neck radiotherapy. The degree of halitosis was determined through the presence of tongue coating, classified according to its dry weight; the salivary flow rate, the BANATM test and HalimeterTM oral measurements. A total of 48 subjects were examined, which were divided into three groups. Group 1 was consisted for patients with good oral health and no complains of bad breath; Group 2 consisted of patients with bad oral health condition and Group 3 comprised individuals who had received head and neck radiotherapy. The concentration of Volatile Sulfur Compounds (VSC) was assessed by means of a portable sulfide monitor (HalimeterTM) and the values were correlated to the resting and stimulated salivary flow rate and to the BANATM test with material collected from the tongue coating and its dry weight. Halitosis was quantified before and after the removal of the tongue coating and the data was analyzed by means of the ANOVA test. The correlation among salivary flow rate, weight of the tongue coating and HalimeterTM oral measurements was evaluated by the Pearson test. The results showed a statistically significant difference between group 1 and 3, and between groups 1 and 2. A relationship between the presence of tongue coating and the VSC levels was also demonstrated. In concerning to the salivary flow rate, there was a statistically significant reduction between group 1 and 3, and group 2 and 3. In the accordance of this study, halitosis can be considered a side effect of radiotherapy, tied with the hyposalivation and deficient oral health.

Os microRNAs regulam uma parte significativa dos genes humanos, dentre os quais genes responsáveis por controlar o ciclo celular. Em estudos anteriores identificou-se maior expressão do miR-196a em amostras de carcinoma epidermoide de cabeça e pescoço (CECP), um dos tipos de tumor mais comuns no mundo, quando comparadas a margens cirúrgicas. Através de estudos de ganho de função, observou-se diminuição da proliferação de queratinócitos orais e de linhagens celulares após a super-expressão do miR-196a. Neste trabalho avaliamos a expressão gênica e de proteínas em linhagem celular em diferentes tempos após super-expressão do microRNA. Identificamos alterações na expressão de genes ativadores e repressores do ciclo e diminuição na expressão de p27, alvo do miR-196a. Observamos que após 48 horas de super-expressão a maioria dos genes repressores do ciclo celular estavam mais expressos, fato possivelmente associado à diminuição da proliferação. Sugerimos que a super-expressão de miR-196a possa ser considerada como uma possível terapia para o tratamento do câncer. / MicroRNAs regulate a proportion of human genes, among which genes responsible for cell cycle control. Squamous cell carcinoma of head and neck (HNSCC) is among the most common malignancies worldwide. In a previous study we identified enhanced expression of miR-196a in HNSCC samples compared with surgical margins and increased expression of miR-196a in HNSCC-derived cell lines compared to oral keratinocytes. Following a gain of function study, there was decreased oral keratinocyte and HNSCC cell line proliferation. In this study, gene expression and protein levels were assessed within 48 hours after overexpression of miR-196a. Changes in gene expression levels of cell cycle activators and repressors was observed, as well as the decreased expression of p27, a gene targeted by miR-196a. At 48 hours following the overexpression of miR-196a most inhibitor genes were more expressed. This fact could be associated with the decrease in cell proliferation after overexpression of miR-196a. We suggest that the overexpression of miR-196a is considered a tool for CECP therapy.

Os avanços na compreensão da biologia das neoplasias de cabeça e pescoço têm aberto novas direções na ciência. As pesquisas estão sendo direcionadas para o desenvolvimento de terapias com alvos moleculares específicos, os quais são úteis tanto na predição dos tratamentos, quanto na seleção de pacientes que podem responder a uma determinada terapia com base nas alterações moleculares dos tumores. As proteínas hnRNP K e SET, recentemente identificadas como superexpressas em câncer de cabeça e pescoço, representam um novo e atrativo alvo terapêutico para esse tipo de câncer. As mitocôndrias também tem sido objeto de estudo, pois participam nos processos de morte celular por apoptose, e estão envolvidas na sobrevivência celular. Neste estudo avaliamos os efeitos in vitro e in vivo em carcinoma oral e em célula não-tumoral, de novos compostos com alvo-específico (hnRNP K e SET) ou com ação na mitocôndria, para aplicação tanto como antitumoral, como citoprotetor. A citotoxicidade dos compostos foi avaliada pelo método de resazurina nas linhagens tumorais de carcinoma de células escamosas de cabeça e pescoço (HN13, HN12, HN6 e CAL27). Os compostos 11 e 17, alvos específicos da proteína hnRNP K, apresentaram baixa citotoxicidade; o peptídeo OP449, alvo específico da proteína SET, e o composto YV-241, com ação na mitocôndria, apresentaram alta citotoxicidade, com valores de IC50 5,11 e 7,77 ?M, respectivamente. OP449 alterou as proteínas reguladas por SET e reduziu a proliferação das células tumorais no modelo de xenoenxerto ortotópico em camundongo BALB/ c nude; os resultados, porém, não foram significativos. A associação de OP449 com FTY720 promoveu um efeito sinérgico significativo (CID<0,7) na linhagem celular HN12 e reduziu os tumores do xenoenxerto no dorso em camundongos BALB/c nude. O composto YV-241 alterou o potencial de membrana mitocondrial das células tumorais e aumentou o número de mitocôndrias, observado por microscopia eletrônica de transmissão e por microscopia confocal, reduziu proteínas envolvidas com vias de sinalização de sobrevivência, proliferação, ciclo celular e angiogênese, e induziu apoptose com o envolvimento da mitocôndria. Além disso, o composto reduziu os tumores do modelo de xenoenxerto. O possível efeito citoprotetor do composto JM-E-H foi observado na linhagem NOK-SI, por meio da regulação da via de sinalização de HIF-1?. Portanto, OP449 + FTY720 e o composto YV-241 apresentam potencial terapêutico contra carcinoma oral, e o composto JM-E-H, potencial efeito citoprotetor / Advances in understanding the biology of head and neck cancer have opened new directions in science. Research is being directed at the development of therapies with specific molecular targets that are useful in predicting treatments or in selecting patients who may respond to a particular molecular therapy based on molecular changes of the tumors. The hnRNP K and SET proteins, recently identified as overexpressed in head and neck cancer, represent a new and attractive therapeutic target for this type of cancer. Mitochondria have also been the object of study, since they participate in the processes of cell death by apoptosis, and are involved in cell survival. In this work we evaluated the in vitro and in vivo effects in oral carcinoma and non-tumor cell of new compounds with specific target (hnRNP K and SET) or with action in mitocondria, for application either as antitumor or cytoprotectant. The cytotoxicity of the compounds was evaluated by the resazurin method in head and neck squamous cell carcinoma cell lines (HN13, HN12, HN6 and CAL27). Compounds 11 and 17, specific targets of hnRNP K protein, showed low cytotoxicity; the peptide OP449, specific target of SET protein, and compound YV-241, acting on mitochondria, showed high cytotoxicity, with IC50 values of 5.11 and 7. 77?M, respectively. OP449 altered SET-regulated proteins and decreased proliferation of tumor cells in the orthotopic xenograft model in BALB/c nude mouse. The results, however, were not significant. The association of OP449 with FTY720 caused a significant synergistic effect (CID <0.7) on HN12 cell line, and decreased the xenograft tumors. The YV-241 compound altered mitochondrial membrane potential of tumor cells and increased the number of mitochondria, observed by transmission electron microscopy and by confocal microscopy, reduced proteins involved with signaling pathways for survival, proliferation, cell cycle and angiogénesis, and induced apoptosis with the involvement of mitochondria. In addition,the compound reduced tumors of the xenograft model. The possible cytoprotective effect of the compound JM-E-H was observed in the NOK-SI lineage through the regulation of HIF-1? signaling pathway. Therefore, OP449 + FTY720 and compound YV-241 show therapeutic potential against oral carcinoma, and the compound JM-E-H, potential cytoprotective effect

Made available in DSpace on 2016-08-10T10:38:45Z (GMT). No. of bitstreams: 1 Cristiana da Costa Luciano.pdf: 1987242 bytes, checksum: 2f3402c7b9c457e5f89de59897be59ea (MD5) Previous issue date: 2012-11-14 / The head and neck cancer is the fifth most common in Brazil, being the most predominant histology type the squamous cell carcinoma. Radiation therapy is a procedure for treatment with the efficacy variable, and may play an important role in controlling tumor growth. Faced with this therapy, the patient is exposed to ionizing radiation that can cause adverse effects resulting cessation of treatment. The objective was to evaluate the association of polymorphisms of TP53 and ATM genes in patients with head and neck cancer with adverse effects on normal tissues presented as a result of radiotherapy. Materials: The DNA was extracted 54 samples of peripheral blood of patients with head and neck cancer, then the fragments of TP53 and ATM were amplified and subsequently sequenced to check for any polymorphism which may be responsible for the radiosensitivity of patients. Statistical analysis was performed using the SPSS 17.0 software. Results and Discussion: In univariate analysis, patients who had experienced adverse effects RT suspended acute low and high grade with RTOG skin (p = 0.012). Those who had a family history of cancer showed higher adverse acute laryngeal RTOG TGI high (p = 0.040). The exchange C> T at position 11322 of TP53 (intron 3), the ATM and TP53 polymorphisms analyzed and the frequency of acute and chronic adverse effects were not significant (p>05). Conclusions: Based on these results is of utmost importance that alternatives are created to predict the dose to be prescribed during radiotherapy, preventing adverse effects and discontinuation of treatment and also providing better tumor control. / O câncer de cabeça e pescoço, no Brasil, é o quinto mais comum, sendo o tipo histológico mais predominante de carcinoma de células escamosas. A radioterapia é uma das modalidades de tratamento com eficácia variável, podendo desempenhar um papel importante no controle do tumor. Diante essa terapêutica, o paciente está exposto a radiações ionizantes que podem causar efeitos adversos gerando a interrupção do tratamento. O objetivo foi avaliar a associação de polimorfismos dos genes TP53 e ATM em pacientes com câncer de cabeça e pescoço com efeitos adversos sobre os tecidos normais apresentados como resultado da radioterapia. Materiais: O DNA foi extraído de 54 amostras de sangue periférico de pacientes com câncer de cabeça e pescoço, em seguida os fragmentos de TP53 e ATM foram amplificados e posteriormente sequenciados a fim de verificar se os polimorfismos poderiam estar associados à radiossensibilidade dos pacientes selecionados. A análise estatística foi realizada utilizando o software SPSS 17.0. Resultados e Discussão: Por meio de análise univariada, pacientes que tiveram a RT suspensa apresentaram efeitos adversos agudo de baixo e alto grau com RTOG de pele (p=0,012). Aqueles que tinham história familiar de câncer apresentaram maiores efeitos adversos agudo de laringe com RTOG TGI alto (p=0,040). A troca C>T na posição 11322 do gene TP53 (intron 3), os polimorfismos de TP53 e ATM analisados e a frequência de efeitos adversos agudos e crônicos não foram significativos para (p>0,05). Conclusões: Diante dos resultados obtidos é de suma importância que alternativas sejam criadas para predizer a dose a ser prescrita durante a radioterapia, prevenindo os efeitos adversos e a interrupção do tratamento e ainda, promovendo melhor controle tumoral.

Neste estudo clínico verificaram-se os efeitos do laser em baixa intensidade na prevenção e tratamento da mucosite oral radio e/ou radioquimioinduzida. Foram selecionados 31 pacientes portadores de câncer em cabeça e pescoço a serem submetidos à radioterapia ou a radioterapia em associação a quimioterapia, os quais foram distribuídos aleatoriamente em três grupos: grupo 1 (controle) tratamento medicamentoso; grupo 2 tratamento medicamentoso e laserterapia diária, a partir do início da ulceração referente à mucosite grau 2 e grupo 3 tratamento medicamentoso e laserterapia diária a ser iniciada imediatamente antes do início da radioterapia. Os parâmetros de irradiação foram: comprimento de onda de 660nm, potência de 100mW, modo de operação contínuo, aplicação pontual, energia de 2J por ponto em 30 pontos pré-determinados, sendo o tempo utilizado de 20s por ponto. O grupo controle recebeu tratamento medicamentoso que consistia no uso de um conjunto de medidas preventivas e terapêuticas para abordagem dos efeitos adversos agudos radioinduzidos. Os resultados foram avaliados, quanto à ocorrência, graus e escore de dor associados à mucosite oral, perda de massa corpórea, uso de sonda nasogástrica, necessidade de internação e interrupção do tratamento oncológico decorrentes da mucosite oral. Os resultados indicam que o protocolo de aplicação do laser em sua forma preventiva é o mais efetivo na prevenção e tratamento da mucosite oral e que seu uso diário contribuiu para o alívio da sintomatologia dolorosa colaborando para melhora da qualidade de vida do paciente oncológico. / In this clinical study verified the effects of low intensity laser in the prevention and treatment of oral mucositis radio and/or chemical induced. Thirty one patients with head and neck cancer were selected before being submitted to cancer exclusive radiotherapy or radio and associated chemotherapy. The patients were distributed into three randomly groups as follows: group 1- (control) conventional medicine treatment; group 2 conventional medicine treatment and daily lasertherapy as soon as grade two oral mucositis appeared; group 3 conventional medicine treatment and daily lasertherapy to be initiated immediately before radiotherapy sessions.The irradiation parameters were: wavelength of 660nm, potency of 100mW, continuous mode, punctual application, 2J energy on thirty pre-determined 30 points, with 20s of exposure per point. The control group received medical treatment which consisted in using a set of preventive and therapeutic approach for acute radiation-induced adverse effects. Results were evaluated observing occurrence and grade of oral mucositis, score of pain, loss of body mass, use of nasogastric sound line, internment and interruption of oncologic treatment due to oral mucositis. The results showed that the preventive protocol as used was the most effective in prevention and treatment of oral mucositis and that its daily application contributed in relieving the painful symptomatology so collaborating to maintain and/or bettering the life quality of oncologic patients.

Introdução: pacientes em tratamento de câncer de cabeça e pescoço podem apresentar alterações vocais decorrentes do tratamento oncológico de radioterapia, quimioterapia e/ou cirurgia. Essas alterações, que podem causar impacto na qualidade de vida, podem estar presentes mesmo que a laringe não seja acometida pelo tumor nem esteja no campo de radiação. Objetivo: avaliar os aspectos vocais e de qualidade de vida relacionados ao tratamento de câncer de cabeça e pescoço não glótico. Método: nos momentos pré e pós-tratamento oncológico, foram avaliados os aspectos vocais de 17 pacientes em tratamento de câncer com localização em cavidade oral, orofaringe, hipofaringe e supraglote por meio de análise perceptivo-auditiva, análise acústica e autoavaliação com questionários de qualidade de vida (UW-QOL e QVV) e de sintomas vocais e sensações na garganta (CPV). Os pacientes foram divididos em dois grupos para análise: submetidos à radioterapia associada à quimioterapia e submetidos à cirurgia e/ou quimioterapia. Resultados: na comparação pré e pós-tratamento, observou-se: pior impacto de qualidade de vida para os domínios de recreação, paladar, saliva e cálculo total após radioterapia; pior impacto de qualidade de vida para o domínio de mastigação após cirurgia e para o domínio dor antes deste tratamento; diminuição da média da frequência fundamental na emissão grave após a radioterapia; aumento dos valores de porcentagem dos parâmetros de perturbação de frequência após a cirurgia. Na comparação entre os grupos foi observado piora para o grupo que realizou radioterapia quanto ao impacto de qualidade de vida nos domínios de atividade, recreação, deglutição, paladar, saliva, humor e ansiedade, no cálculo total, na questão que compara o momento atual com o momento antes de apresentar o câncer e na questão de classificação da qualidade de vida geral, além de maior frequência de sintoma de rouquidão e sensações de tosse com catarro, secreção/catarro na garganta e garganta seca. Na análise perceptivo-auditiva comparativa da qualidade vocal pré e pós-tratamento, os juízes consideraram que 57 % dos casos apresentou piora vocal no momento pós-radioterapia e 31% no momento pós-cirurgia, sem diferenças estatisticamente significativas. Conclusão: alguns aspectos vocais e de qualidade de vida apresentaram-se alterados após o tratamento de câncer de cabeça e pescoço não glótico, sendo estas alterações mais presentes após o tratamento de radioterapia comparado com os demais tratamentos oncológicos. / Introduction: patients being treated for head and neck cancer may present vocal alterations caused by cancer radiotherapy, chemotherapy and/or surgery. These changes, which can impact on quality of life, may be present even if the larynx is not affected by the tumor nor is the larynx in the radiation field. Objective: To evaluate the vocal and life quality aspects related to the treatment of head and neck cancer not glottis. Method: in pre- and post-treatment cancer, they were evaluated the vocal aspects of 17 patients undergoing cancer treatment with location in the oral cavity, oropharynx, hypopharynx and supraglottic through perceptual analysis, acoustic analysis and self-assessment with quality questionnaires of life (UW-QOL and QVV) and vocal symptoms and sensations in the throat (CPV). The patients were divided into two groups for analysis: one group undergoing radiotherapy combined with chemotherapy and the other underwent surgery and/or chemotherapy. Results: when comparing pre and post-treatment, it was observed: worst impact quality of life for recreation, taste, saliva domains and total calculation after radiotherapy; worst impact quality of life for the chewing domain after surgery and the pain domain before this treatment; decrease in average fundamental frequency in low range voice issue after radiotherapy; increase in the percentage values of the disturbance frequency parameters after surgery. In the comparison between groups was observed worsening for the group that underwent radiotherapy as the impact of quality of life in the domains of activity, recreation, swallowing, taste, saliva, mood and anxiety, in total calculation, the question that compares the current time with the time before submitting the cancer and the issue of the general quality of life rating and greater frequency of symptom hoarseness and cough with phlegm sensations, secretion / phlegm in the throat and dry throat. In the comparative in the perceptual analysis of voice quality pre and post-treatment, the court ruled that 57% of the cases presented vocal worsening post-radiotherapy time and 31% in the post-surgery time without statistically significant differences. Conclusion: some vocal and life quality aspects have to altered after treatment of head and neck cancer not glottis, being these alterations more present after radiotherapy compared with other cancer treatments.

Introdução: O câncer de cabeça e pescoço vem aumentando sua incidência, sendo o quinto tipo de câncer mais comum e a sexta causa de morte por câncer no mundo, portanto é causa importante de morbimortalidade da população. O entendimento dos fatores de risco como tabagismo, etilismo e infecção pelo papilomavirus (HPV) é de fundamental importância, tanto para ações de prevenção e controle da doença, bem como avaliação de fatores prognósticos e terapêuticos. Objetivos: O presente estudo visa analisar as características clínicas, buscar o principal sítio anatômico tumoral e avaliar os fatores prognósticos entre os pacientes tabagistas ou não, e portadores ou não do HPV, que foram tratados cirurgicamente. Casuística e Métodos: Foram analisadas as variáveis clínicas e prognósticas dos grupos de pacientes tabagistas, não tabagistas, HPV negativo e HPV positivo. A amplificação do DNA viral, detecção e genotipagem do HPV se deu através da extração do DNA total a partir de blocos parafinados. Resultados: A amostra total foi composta por 399 pacientes, sendo que 266 eram tabagistas, 33 não tabagistas, e desses últimos, cinco eram HPV positivo. Houve diferença estatística em relação sexo, sendo que para o tabagista o predomínio foi do sexo masculino e para o HPV positivo foi o feminino. O paciente tabagista apresentou-se mais jovem (média 57,9 anos) que o não tabagista (média 64,1 anos). O sítio anatômico mais comum para os tabagistas foi a laringe e para o HPV positivo foi a cavidade oral e orofaringe. O tempo livre de doença foi de 63,6 meses para os tabagistas, 31,3 meses para os não tabagistas e 29,3 meses para os HPV positivo, havendo diferença quanto às curvas de sobrevidas livre da doença dos fumantes e não fumantes. Dos pacientes que foram á óbito pela neoplasia, o paciente HPV positivo foi o que apresentou menor tempo de sobrevida. Não houve diferença estatística entre os grupos quanto às curvas de sobrevidas global e da doença. Discussão: Apesar da literatura mostrar que o paciente não tabagista e o paciente HPV positivo geralmente serem mais jovens e apresentarem melhor prognóstico do que os típicos pacientes tabagistas, isso não foi observado nesse estudo, pois o paciente não tabagista apresentou-se com mais idade ao diagnóstico da doença e com pior prognóstico que o tabagista, e o paciente HPV positivo apresentouse com menor tempo até o óbito pela doença quando comparado ao paciente tabagista. Conclusões: O câncer nos pacientes tabagistas acometeu mais o sexo masculino, localizou-se principalmente na laringe e apresentou melhor prognóstico quando comparado aos não tabagistas. O câncer nos pacientes HPV positivo acometeu mais o sexo feminino e localização tumoral foi principalmente a cavidade oral e a orofaringe. / Introduction: Head and Neck cancer has been increasing its incidence, and it is the fifth most common type of cancer and the sixth leading cause of cancer death in the world, so it is an important cause of morbidity and mortality of the population. Understanding the risk factors such as smoking, alcohol use and papillomaviruses (HPV) infection is of fundamental importance, both for prevention and control of disease like for assessment of prognostic and therapeutic factors. Objectives: This study aims to analyze the clinical, seek the main site anatomical of the tumor and evaluate the prognostic factors among smokers or not, with the presence or absence of the HPV, which were surgically treated. Methods: The variables clinical and prognostic were analyzed in the groups of smokers, nonsmokers, HPV negative and HPV positive. The amplification of viral DNA, detection and genotyping of the HPV were made through the extraction of total DNA from paraffin blocks. Results: The total sample consisted of 399 patients, 266 were smokers, 33 nonsmokers, and this latter group had five patients HPV positive. There was statistical difference regarding sex, for smokers the prevalence was male and for HPV positive was female. The smoker patient was younger (mean 57.9 years) than the non-smoker (mean 64.1 years). The most common anatomic site for the smokers was the larynx and for the HPV positive was the oral cavity and oropharynx. The diseasefree interval was 63.6 months for smokers, 31.3 months for nonsmokers and 29.3 months for HPV positive, with difference between free disease survival curves of the smokers and nonsmokers. Of the patients who died by disease, HPV positive patient showed the shorter survival. There was no statistical difference between the groups in terms of overall survival curves and of the disease. Discussion: Although the literature show that the nonsmoker patient and HPV positive patients are generally younger and present a better prognosis than the typical smokers, this was not observed in this study because the nonsmoker patient presented with more age at diagnosis of disease and a worse prognosis than the smoker, and the HPV positive patient presented with shorter survival of the disease when compared to smoking patients. Conclusions: The cancer in smokers affected more males, was located mainly in the larynx and showed better prognosis when compared to nonsmokers. The cancer in patients positive HPV affected more females and tumor location was mainly oral cavity and oropharynx.

A preservação da mobilidade mandibular é um aspecto fundamental para a manutenção da qualidade de vida de pacientes oncológicos submetidos à radioterapia. Este estudo avalia o tempo mínimo de 1 ano decorrido após a realização do tratamento radioterápico, dose e aparelho radioterápico empregados para terapêutica, utilização de sonda nasogástrica e tipo de alimentação, acompanhamento e fisioterapia miofuncional. Integraram este estudo pacientes da Fundação Oncocentro do Estado de São Paulo. Os dados clínicos foram coletados nos respectivos prontuários e as mensurações da amplitude dos movimentos mandibulares de máxima abertura, protrusão e lateralidade direita e esquerda foram registradas em sessão clínica com auxílio de um paquímetro digital. Observou-se que: os gêneros masculino e feminino respondem de modo semelhante na avaliação do comprometimento da mobilidade mandibular; a radioterapia associada à oncocirurgia é a opção terapêutica mais utilizada para o tratamento oncológico de tumores da cavidade oral; o tempo decorrido para a detecção da alteração da mobilidade mandibular é bastante longo, impondo-se medidas preventivas e mesmo interceptadoras mais precoces como o acompanhamento e tratamento com terapia miofuncional; a dose de irradiação e o tipo de aparelho radioterápico empregados no tratamento de pacientes oncológicos são determinantes da alteração da mobilidade mandibular; a utilização de sonda nasogástrica e a dieta pastosa potencializam o efeito da radioterapia no que diz respeito à mobilidade mandibular. / The preservation of mandibular mobility is a fundamental aspect for the maintenance of the quality of life of cancer patients undergoing radiotherapy. This study evaluates the minimum time of 1 year after radiotherapy, dose and radiotherapy used for therapy, use of nasogastric tube and type of feeding, monitoring and myofunctional physiotherapy. Patients at the São Paulo Oncocentro Foundation were enrolled in this study. The clinical data were collected in the respective charts and the measurements of the amplitude of the mandibular movements of maximum opening, protrusion and right and left laterality were recorded in a clinical session with the aid of a digital caliper. It was observed that: the masculine and feminine genera respond similarly in the evaluation of mandibular mobility impairment; radiotherapy associated with oncosurgery is the most used therapeutic option for the oncological treatment of tumors of the oral cavity; the elapsed time to detect the change in mandibular mobility is quite long, imposing preventive and even earlier interceptive measures such as monitoring and treatment with myofunctional therapy; the dose of irradiation and the type of radiotherapy used in the treatment of cancer patients are determinants of the mandibular mobility alteration; the use of nasogastric tube and pasty diet potentiate the effect of radiotherapy with respect to mandibular mobility.

Oncolytic viruses can be genetically modified to limit their replication in normal cells rendering them a cancer specific treatment. In addition, they can induce a 'danger signal' in the form of pathogen- and damage-associated molecular patterns leading to anti-tumour immunity. Furthermore, they can be armed with various immunomodulatory molecules to further enhance anti-tumour immunity. In this project I aim to exploit these qualities to develop a translatable cancer vaccine. Virus-infected cancer cells were injected subcutaneously in a prime/boost regimen. Dying cancer cells will release the required danger signal leading to dendritic cell activation and cross-presentation of tumour associated antigens to T cells to elicit an anti-tumour immune response. Our results in the murine pancreatic cancer model showed that vaccination with virusinfected DT6606 cells induced tumour specific immunity capable of protecting vaccinated animals against re-challenge with tumour cells. The highest level of interferon gamma production, a surrogate marker of anti-tumour immunity, was achieved when animals were primed with adenovirus-infected cells. There was no significant difference between various boost groups. To enhance the safety of the proposed protocol a secondary treatment was introduced to arrest the proliferation of tumour cells prior to injection. Our results confirmed that secondary treatment with mitomycin does not affect the induction of tumour specific immunity and it does not affect the release of pathogen-associated molecular patterns in the form of viral proteins and DNA. To test our vaccination regimen in head and neck squamous cell carcinoma (HNSCC) we develop a clinically relevant mouse model using SCC7, B4B8 and LY2 cells to replicate various clinical scenarios including locally advancing disease and post excision locoregional recurrence. Vaccinating mice with HNSCC cells pre-infected with our recently developed tumour-targeted triple-deleted adenovirus (AdTD) resulted in a cell-specific antitumour immune response. In addition, it resulted in an increase in effector memory T-cells of both CD4+ and CD8+ phenotypes. Efficacy studies showed our vaccination can significantly slow down the growth rate of tumours in locally advancing disease. This led to increase survival of the vaccinated mice although it did not reach statistical significance. To further enhance the efficacy of our vaccination regimen, we aimed to increase T cell trafficking to the tumour site. CCL25 is a gut homing chemokine. Priming T cells in the presence of CCL25 will lead to upregulation of the surface expression of α4β7 integrin. The latter is a ligand of MAdCAM-1, a cell adhesion molecule highly expressed in the gut and pancreatic tumours. The α4β7/MAdCAM-1 interaction results in preferential homing of activated T cells to these organs. We hypothesised that vaccinating mice with pancreatic tumour cells pre-infected with a CCL25-armed adenovirus will lead to increased T cell trafficking to pancreatic tumours leading to enhanced efficacy. Although we achieved encouraging results in our pilot experiment, we did not detect any significant increase in α4β7 expression once we added a secondary treatment to the vaccination protocol. Similarly, efficacy experiments in the pancreatic cancer transgenic KPC mice did not show any difference in survival between AdTD-CCL25 and the control virus although both groups showed a trend towards increased survival compared to naïve mice. In conclusion, Virus-infected cancer cell vaccine is a potentially promising immunotherapeutic strategy that can be combined with traditional cancer therapies to increase survival of HNSCC and pancreatic cancer patients.

HOXA7 belongs to a family of homeobox transcription factors that are master regulators of cell differentiation, morphogenesis during embryonic development and cell proliferation. Dysregulation and non-nuclear localization of these proteins play a role in a large number of solid tumours, with reports of significant upregulation of HOXA7 in oral dysplasia. It is unclear whether HOXA7 induction in solid tumours is causative or if it is a result of oncogenic changes. In this thesis we studied its effect on cell differentiation, growth, stemness, cell migration, EMT and cell senescence. The main hypothesis was that HOXA7 regulated keratinocyte differentiation through the regulation of activator protein 1 (AP-1), a keratinocyte specific activator of differentiation. We also hypothesised that HOXA7 increased the proliferation rate in keratinocytes. In an AP-1 reporter assay in HEK293 cells, HOXA7 was shown to decrease AP-1 activity significantly. The inactivation of AP-1 was not due to inactivation of PKC, as HOXA7 did not interfere with the activation of the kinases in HEK293. More specifically, we reported a very significant repression of c-Jun and JunD promoter activity in the presence of ectopic HOXA7 in HEK293 cells. We further showed that this mechanism might also be applicable in keratinocytes, as HOXA7 inhibited the transcription of AP-1 subunits of both the Jun and Fos family in skin keratinocytes. Furthermore, we showed transcriptional repression of four differentiation markers and a downregulation of K1 and FLG protein in transduced NEB-1 monolayers as well as K1 suppression in HaCaT cells. The organotypic cultures revealed a downregulation of K1, K10, and filaggrin in stratified HaCaT cells by HOXA7. There was however no downregulation in oral keratinocytes. These observations taken together suggested that HOXA7 repressed the synthesis of AP-1 units in skin keratinocytes, which would have resulted in reduced quantities of AP-1 and therefore lower activity. Contrary to previous reports, we observed no positive involvement of HOXA7 in keratinocyte proliferation, EMT or migration. There was however an indication of cell-type specific MET and induced cell senescence. Based on our results we propose a cell-type specific role of HOXA7 as an antagonist of AP-1 transcription in skin keratinocytes, and a possible direct binding of HOXA7 to c-Jun and JunD promoters.