There are unmet needs in evaluating treatment response of hepatocellular carcinoma in research protocols. Early predictors, such as imaging biomarkers, could allow for earlier judgment of treatment effect. Currently RECIST is the most widely accepted criterion in clinical trials. A modified RECIST (mRECIST) criterion was developed to take into account the unique imaging characteristics of HCC lesions. Much discussion has occurred regarding linear measurements and their appropriateness for evaluating change in tumor burden over time. The simplicity of currently accepted criteria differs with the increasing sophistication of imaging techniques. Tumor volume change on 3D imaging can provide insight into actual action of treatment rather than an estimate of action as shown by linear and bi-dimensional measurements. It was the aim of this study to determine whether linear, bi-dimensional, and volumetric percent changes of HCC lesions, in both the arterial and portal venous phases, are significantly comparable. 27 HCC lesions (identified on 25 subjects) were measured at two timepoints by each method on 3D GRE MRI scans in both phases. Percent change was calculated per lesion for each measurement type in both the arterial and portal venous phases. Signed rank tests, paired t tests, and comparison of change tests were run to evaluate the data. Significant differences between the percent changes of linear measurements versus volumetric measurements were observed using a Wilcoxon signed-rank test which showed p = 0.0000. A simple correlation assessment showed positive correlations for all measurements, with the lowest being correlations 0.8679 for the arterial linear percent change versus the arterial volumetric percent change and 0.8434 for the portal venous linear percent change versus the portal venous volumetric percent change. Differences between percent changes of linear versus bi-dimensional measurements and bi-dimensional versus volumetric measurements were significant as well (Linear versus bi-dimensional p = 0.0001, bi-dimensional versus volumetric p = 0.0004). To conclude, the differences in the percent changes when comparing the measurement types are statistically significant, particularly when comparing linear and volumetric measurements. Establishing a reproducible volumetric criterion could lead to improvements in the implementation of clinical trials.

Thesis (Ph.D.)--Boston University / Ehrlich ascites carcinoma lactic acid dehydrogenase was isolated from an eleven-day old tumor by acid precipitation, ammonium sulfate fractionation, and chromatography on DEAE cellulose. Electrophoretic analysis indicated that the final enzyme preparation and the ammonium sulfate fraction contained a single isoenzyme, that is, one of the five possible forms of lactic acid dehydrogenase two of which are tetramers of a single but different protein while the other three are tetramer mixtures of both proteins (i.e. hybrid enzymes). Ultracentrifugal analysis indicated that the final enzyme preparation was composed of two major components with sedimentation rates of 7.3 S and 1.9 S. The enzymatic activity was associated only with the 7.3 S component. The apparent loss of enzymatic activity in 0.1 M phosphate buffer pH 7.0 and the magnitude of the value of the 1.9 S component indicated that this represented the subunit of the enzyme. [TRUNCATED]

Oral squamous cell carcinoma (OSCC) is the sixth most common cancer worldwide. It is a result of numerous aetiological factors such as genetic predisposition, smoking, excessive alcohol consumption and viruses such as the human papilloma virus. Due to late diagnosis it has a high mortality and morbidity rates which has remained unchanged over the last 5 decades. Currently no screening is available for high risk patients for better monitoring. Diagnosing OSCC relies on histopathology of biopsy tissue, reviewed for dysplasia and advancing lesions. Although the technique has been used for decades for successful diagnosis it fails to identify the molecular signature of OSCC which appears much before the visual signs. It also falls short in predicting the malignant transformation of pre-malignant oral lesions. Identifying the molecular and genetic changes leading to OSCC lesion will aid in more specific (quantitative) and early diagnosis of the disease reducing the financial burden of treating late-stage OSCC patients on the healthcare system. This study focuses on developing new adjuncts which can be used alongside histopathology for early diagnosis. There is a need to monitor high risk patients through non-invasive methods causing less patient discomfort. We therefore explored the potentials of exosomes which are extracellular vesicles secreted by normal and tumour cells. They can be isolated from body fluids such as blood and saliva. In cancer biology exosomes offer both diagnostic and therapeutic advantage. Their involvement in cell-cell communication indicates their influence in tumour development, progression, metastasis and therapeutic efficacy. Exosomes released by cancerous cells carry numerous biomarkers, which are passed on to healthy cells via microenvironment, causing stromal and angiogenic activation along with immune escape. In this study exosomes were successfully isolated from body fluids (blood, saliva and plasma) and cell line supernatant through ultracentrifugation and characterised by visual and particle size quantification techniques including Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM), Zetasizer and Nanosight Tracking Analysis (NTA). Exosomal specific membrane proteins were identified through Western blotting. 5 We report the presence of a potential protein biomarker located exclusively on the outer membrane of cancer exosomes. Since body fluids consist of a heterogeneous population of exosomes derived from multiple cell types, such surface biomarker can potentially be used to isolate OSCC exosomes. Characterisation of exosomal mRNA cargo was done using Agilent Bioanalyzer (for RNA quantity and quality assurance) and reverse transcription-quantitative PCR (RT-qPCR; for gene specific quantitation). Functional significance of exosomes was studied by transfecting normal oral keratinocyte cells with self and cancer-derived exosomes. Through gene-expression microarray and subsequent RT-qPCR verification, we report a panel of differentially expressed genes involved in essential cellular functions being modulated by exosome transfection. A previously developed molecular diagnostic system by our research group called quantitative malignancy index diagnostic system (qMIDS) based on FOXM1 oncogene and its downstream targets was validated on archival formalin fixed paraffin embedded OSCC patient biopsy samples. We report that qMIDS index successfully correlates with the disease stages including dysplasia, tumour and lymph node metastasis. Furthermore, through meta-analysis of 8 OSCC microarray studies we identified a panel of six genes including PLAU, FN1, CDCA5, CRNN, CLEC3B and DUOX1 (q6) which are able to identify two clinically distinct sub-groups of OSCC patient population. Through RT-qPCR the expression of q6 biomarkers was established in 100 OSCC biopsy samples. This information can be of immense importance in developing personalized treatment strategies based on the molecular makeup of the presenting tumour.

O objetivo deste estudo foi avaliar a proliferação celular de ceratinócitos no dorso e ventre de língua de ratos submetidos à ingestão de etanol a 5%, por meio da técnica de AgNOR, sendo um estudo experimental randomizado, em paralelo, cego, controlado. Foi analisado o epitélio lingual (Dorso e Ventre) de 30 ratos, divididos em 2 grupos (Teste e Controle). Foram quantificadas a média e a percentagem de AgNOR das células das camadas basal e suprabasal, comparando os grupos, pelo período de 9 semanas. Os resultados mostram que houve diferença estatisticamente significante no grupo teste (etanol), com uma maior atividade proliferativa para o ventre, quando p AgNOR > 3 e p AgNOR > 4. Conclui-se que a exposição ao etanol a 5% provoca alterações de proliferação no modelo estudado. / The aim of this study was to evaluate the cellular proliferation of keratinocytes in the dorsal and ventral surface of tongue of rats after ingestion of ethanol to 5% by AgNOR technique, being an experimental randomized, parallel, blinded, controlled. Lingual epithelium was examined (dorsal and ventral) of 30 rats were divided into two groups (Test and Control). Were quantified and the average percentage of AgNOR cell basal and suprabasal layers, comparing the groups for a period of nine weeks. The results show a statistically significant difference in the test group (ethanol) with a higher proliferative activity to the womb, when AgNOR p> 3 AgNOR p> 4. It is concluded that exposure to 5% ethanol causes changes in proliferation in this model.

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O carcinoma colorretal (CCR) está entre os três tipos mais frequentes e mortíferos de câncer do mundo. O tratamento de primeira escolha depende do estágio tumoral, sendo que o regime quimioterápico mais utilizado é o FOLFOX (5-fluoruracila + oxaliplatina + leucovorina). Apesar do amplo uso, o mecanismo de ação deste protocolo, especialmente considerando o período entre ciclos de tratamento, bem como os mecanismos envolvidos na frequente resistência de células de CCR ao tratamento são pouco conhecidos. Aqui nós avaliamos a resposta de células de CCR expostas ao tratamento com 5-fluoruracila (5FU) e oxaliplatina (OXA) em perfil semelhante ao tratamento clínico, nas linhagens de CCR humano HCT116 e HT29, em prol de entender os mecanismos de ação e de resistência ao tratamento. Para isso, as células foram tratadas com os quimioterápicos de maneira isolada ou combinados por 2 dias, seguido do replaqueamento em meio livre de droga por 15 dias, mimetizando o regime de tratamento clínico. Foram realizadas análises da proliferação celular e ciclo celular, além de análises dos mecanismos celulares que vêm sendo mostrados como envolvidos na sensibilidade e resistência de células tumorais a compostos citotóxicos, tais como apoptose, senescência e autofagia. Não houve efeito aditivo citotóxico para a combinação de 5FU e OXA a curto prazo (48h), porém, a longo prazo houve um efeito aditivo importante para o co-tratamento quando comparado aos tratamentos isolados. Este efeito aditivo foi mediado pela indução de apoptose (principalmente pela 5FU) e senescência (principalmente pela OXA). Apesar deste efeito, as células de todos os tratamentos retomaram o crescimento bem como mantiveram capacidade clonogênica a partir do dia 7 após o tratamento. Quando nós avaliamos a autofagia nas células sobreviventes ao tratamento, encontramos um aumento transitório, com pico entre os dias 3 e 7, sendo que um perfil semelhante foi observado para o aumento de ROS, com pico entre os dias 5 e 7. A inibição racional da autofagia no período de ativação máxima do mecanismo reduziu de maneira intensa o número de células e a clonogenicidade das células sobreviventes. Dessa forma, a inibição da autofagia em combinação com 5FU e OXA pode potencializar o efeito do tratamento e reduzir a resistência das células de CCR ao tratamento. / Colorectal carcinoma (CRC) is among the three most frequent and deadly types of cancer in the world. The first-choice treatment depends on the tumor stage, and the most commonly used chemotherapy regimen is FOLFOX (5-fluorouracil + oxaliplatin + leucovorin). Despite the wide use, the mechanism of action of this protocol, mainly considering the interval between two cycles of chemotherapy, as well as the mechanisms involved in the frequent resistance of CRC cells to treatment are poorly understood. In this project, we evaluated the response of HCT116 and HT29 human CRC cells exposed to 5-fluoruracil (5FU) and oxaliplatin (OXA) in a profile similar to clinical treatment. Cells were treated with the chemotherapeutics alone or in combination for 2 days, followed by the replating in drug-free medium for 15 days, mimicking the clinical treatment regimen. Analysis of cell proliferation and cell cycle were performed, as well as the analysis of the cellular mechanisms that have been shown as being involved in the sensitivity and resistance of tumor cells to cytotoxic compounds, such asapoptosis, senescence and autophagy. There was no cytotoxic additive effect for the combination of 5FU and OXA in the short-time (48h), but in the long term it was observed an important additive effect for co-treatment when compared to the isolated treatments. This additive effect was mediated by the induction of apoptosis (mainly by 5FU) and senescence (mainly by OXA). Despite this effect, cells from all treatments resumed growth as well as maintained the clonogenic capacity from day 7 after treatment. When we evaluated autophagy in treatment-surviving cells, we found a transient increase, with a peak between days 4 and 6; a similar transient increase was observed for reactive species, peaking between days 6 and 8. The rational inhibition of autophagy in the period of maximal activation of the mechanism greatly reduced the number of cells and the clonogenicity of the surviving cells. Thus, the rational inhibition of autophagy in combination with 5FU and OXA may potentiate the effect of the treatment and reduce the resistance of CRC cells to the treatment.

Le foie est un organe essentiel à la vie chez tous les mammifères. C’est un organe central du métabolisme énergétique et de la détoxification des substances xénobiotiques auxquelles l’individu est exposé. Le foie est la cible d’agressions diverses, telles que les virus, l’alcool, les substances chimiques présentes dans l’alimentation ou l’environnement. Il peut également subir destransformations pathologiques profondes, lors du diabète ou de l’obésité par exemple.La stéatose hépatique, caractérisée par une accumulation de triglycérides sous forme de vésiculesgénérant une réponse inflammatoire, est connue depuis de nombreuses années. Son étude a permisde définir un modèle en deux étapes (« two hits ») indispensables à la genèse d’une stéatohépatite ou NASH. La première est l’accumulation de lipides, la seconde consiste en la genèse d’un stress oxydant et la libération de cytokines. La NASH est une des conséquences pathologiques du syndrome métabolique au cours duquel une résistance des tissus à l’insuline se développe.Récemment, la composition des lipides accumulés dans la NASH a été décrite et montre la présence de cholestérol libre et de différents métabolites des acides gras dont la toxicité est grande mais variable. De façon surprenante, une nouvelle hypothèse tend à émerger quant aux rôles protecteurs de certaines catégories de lipides. En effet, le stockage des triglycérides sous forme de vésicules pourrait être un mécanisme de survie cellulaire (Neuschwander-Tetri, 2010). Il s’agirait principalement d’une tolérance à la mort cellulaire par nécrose ou apoptose. Dans ce contexte,l’activation de l’autophagie serait capitale et la nécrose ne serait plus un mécanisme non contrôlé,mais au contraire un système finement régulé.Des données expérimentales récentes suggèrent l’existence d’un réseau complexe d’interactions moléculaires qui lient, dans la NASH comme dans le cas de la cancérogenèse, le métabolisme énergétique, la réponse inflammatoire systémique et tissulaire et des altérations subcellulaires, telles que les lésions des mitochondries et du réticulum endoplasmique.Nous avons utilisé le cas particulier du préconditionnement ischémique, une technique chirurgicale qui consiste, grâce à de courtes périodes d’occlusion vasculaire avant l’ischémie, à conférer au tissu une protection contre les lésions d’ischémie/reperfusion (I/R), pour étudier les mécanismes de survie mis en place par les hépatocytes stéatosiques au cours d’un stress d’I/R. Dans deux contextes différents, celui d’une ischémie chaude au cours d’une hépatectomie partielle et celui d’une ischémie froide au cours de la transplantation hépatique, nous avons montré que l’autophagie peut jouer un rôle central dans la protection des hépatocytes stéatosiques. Cependant, il est envisageable qu’un dysfonctionnement de l’autophagie pourrait conduire à la genèse d’altérations cellulaires comme une instabilité génomique, caractéristique de la transformation cancéreuse. L’équilibre entre la survie et la mort cellulaire dépend donc de l’intégration de cette signalisation complexe, qui concerne l’état énergétique de la cellule, la réponse aux stress transitoires et l’adaptation aux stress chroniques. Dans ce contexte, l’autophagie semble jouer un rôle central dans l’intégration de la réponse aux stress (Kroemer et al 2010), ce qui pourrait favoriser directement ou indirectement la transformation cancéreuse d’une cellule.L’amélioration de la compréhension des mécanismes impliqués dans la tolérance et la survie des hépatocytes chargés de lipides en réponse à un stress inflammatoire, ischémique ou du réticulum endoplasmique semble donc essentielle. Elle permettrait en effet la mise en place de nouvelles stratégies thérapeutiques qui pourraient améliorer la prise en charge des patients, augmenter le nombre de greffons disponibles pour les greffes, et la prévention des risques cancérogènes pour le foie. / Liver is a an essential to life organ in all mammals. It plays a central role in energy and drug metabolism. Liver is constantly challenged by damaging compounds such as viruses, alcohol and chemicals from food intake or from the environment. It can also undergo some deep pathological transformations, e.g. in diabetes or obesity. Liver steatosis has been known for many years, it is defined as an accumulation of triglycerides vesicles generating an inflammatory response in hepatocytes. A « two step hypothesis » has been proposed for the genesis of Non Alcoholic Steatohepatitis (NASH). The first step is the fat accumulation, the second step involves the generation of an oxidative stress and the release ofcytokines. NASH is one of the pathological consequences of metabolic syndrome, when insulin resistance occurs in the tissues.The composition of accumulated fat in NASH has been recently described and reveals the presence of free cholesterol and different fatty acids metabolites with a high but variable toxicity. Surprisingly, a new hypothesis tends to emerge about the protective effects of some types of lipides.Triglyceride storage in vesicles could indeed be a survival mechanism for cells (Neuschwander-Tetri, 2010). It is assumed that it would mainly result in an tolerance to cell death by necrosis orapoptosis. In this context, (activation of) autophagy would play a key-role and necrosis, usually an uncontrolled mechanism, would become accurately regulated. Similarly to oncogenesis, recent experimental data in NASH suggest that energy metabolism,systemic and tissular inflammatory response and subcellular alterations such as impaired mitochondria and ER are connected in a complex network of molecular interactions. Ischemic preconditioning (IP) is a surgical technique consisting of brief periods of vascular occlusion which confer protection against subsequent ischemia/reperfusion via endogenous protective mechanisms. We investigated the survival mechanisms set up by steatotic hepatocytes during I/R, with or without IP. In the following two situations, warm ischemia during partial hepatectomy and cold ischemia during liver transplantation, we pointed out that autophagy can play a central role in steatotic hepatocytes protection. However, an autophagy dysfunction might result in the generation of cellular impairments such as genomic instabilities, typical features of oncogenic transformation. Therefore, the balance between cell survival or death depends on the integration of a complex signaling, taking into account the cellular energetic state, the cell response to transient stress and its adaptation to chronical stress. In that context, autophagy seems to play a central role in the integration of stress response (Kroemer et al Mol Cell 2010), which could promote, directly or indirectly the malignant cell transformation.Therefore, it seems essential to improve the understanding of mechanisms involved in tolerance and survival of lipid-full hepatocytes in response to an inflammatory, ischemic or ER stress. Indeed, this would help developing new therapeutical strategies to improve patients care, increase the number of available grafts for transplants, and prevent cancer risks in liver.

Orientador: Cláudia Maria Navarro / Resumo: A queilite actínica (QA) é causada pela exposição crônica à radiação ultravioleta (UV), afetando principalmente o lábio de pessoas de pele clara, com 50 anos ou mais, sendo característica a perda da delimitação mucocutânea. A QA mesmo possuindo potencial para malignização ainda não é considerada uma desordem potencialmente maligna oral (DPMO), e isto se deve à falta de estudos com casuísticas consistentes. O objetivo desse estudo foi a caracterização da QA como DPMO mediante estudo do índice de transformação maligna e correlação clinicopatológica. Foram avaliados 182 prontuários de pacientes com QA atendidos no Serviço de Medicina Bucal (SMB) do Departamento de Diagnóstico e Cirurgia, da Faculdade de Odontologia de Araraquara - UNESP, no período de 1990 a 2018. Os dados foram transferidos para um banco de dados do programa Epi Info 7.2.2.6 software aberto. A comparação de dados paramétricos foi realizada por meio do teste Qui - Quadrado e o teste Exato de Fisher, teste T Student e o teste de Mann-Withney. Em testes não paramétricos foi utilizado o teste Anova One way com pós - teste de Tukey. Foi realizado também teste de Regressão categórica linear utilizando o software IBM SPSS Statistics 20, p <0,05 foi considerado significante. Analisando a amostra de 182 pacientes, sendo 71,97% sem carcinoma espinocelular (CEC) e 28,02% com CEC, pode-se traçar o perfil do paciente com QA. Pudemos concluir que as QA acometem principalmente o lábio inferior de homens brancos com mais de 50... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Actinic cheilitis (AC) is caused by chronic exposure to ultraviolet (UV) radiation, affecting mainly the lip of white people aged 50 or older, with loss of mucocutaneous delimitation. The AC even having potential for malignancy it is still not considered a potentially malignant oral disorder (PMOD), and this is due to the lack of studies with consistent sample. The objective of the present study was to characterize the AC as PMOD by studying the index of malignant transformation and clinicopathological correlation. A total of 182 clinical files of patients with AC was attending at the Oral Medicine Service (OMS) of the Department of Diagnosis and Surgery, University of Dentistry of Araraquara - UNESP, in the period 1990 to 2018. The data were transferred to a database of the Epi Info 7.2.2.6 open software program. The parametric data were compared using the Chi - Square test and the Fisher Exact test, Student 's T test and the Mann - Withney test. In non - parametric tests the Anova One way test was used with Tukey post - test. A linear categorical regression test using the IBM SPSS Statistics 20 software was also performed, p <0.05 was considered significant. Analyzing the sample of 182 patients, 71.97% without squamous cell carcinoma (SCC) and 28.02% with SCC, the profile of the patient with AC can be traced. We can conclude that the AC affects mainly the lower lip of white men over 50 years old who suffer chronic exposure to the sun without protection. Although UV radiatio... (Complete abstract click electronic access below) / Mestre

Cheung Ka Wai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 83-93). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Publications --- p.ii / Awards --- p.iii / List of abbreviations --- p.iv / List of figures --- p.vi / List of tables --- p.vii / Abstract in English --- p.viii / Abstract in Chinese --- p.ix / Table of Contents / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- literature Review / Chapter 2.1 --- Anatomy and Physiology of Endometrium --- p.2 / Chapter 2.2 --- Endometrial cancer --- p.5 / Chapter 2.2.1 --- Epidemiology --- p.6 / Chapter 2.2.2 --- Etiologies and Risk Factors --- p.7 / Chapter 2.3 --- Pathology --- p.11 / Chapter 2.3.1 --- Grading of endometrial cancer --- p.14 / Chapter 2.3.2 --- Staging of endometrial cancer --- p.14 / Chapter 2.4 --- Prevention and Treatment --- p.16 / Chapter 2.5 --- Molecular alterations in endometrial cancer --- p.17 / Chapter 2.5.1 --- Genetic alterations in endometrial cancer --- p.18 / Chapter 2.5.1.1 --- Oncogene activation --- p.19 / Chapter 2.5.1.2 --- Tumor suppressor gene inactivation --- p.20 / Chapter 2.5.1.2.1 --- Mutation and loss of heterozygosity of tumor suppressor genes in endometrial cancer --- p.22 / Chapter 2.5.2 --- Epigenetic alterations --- p.27 / Chapter 2.5.2.1 --- CpG islands methylation --- p.29 / Chapter 2.5.2.2 --- de novo methylation --- p.29 / Chapter 2.5.2.3 --- Detection of gene promoter hypermethylation --- p.31 / Chapter 2.5.2.4 --- Epigenetic alteration in endometrial cancer --- p.31 / Chapter 2.5.2.5 --- Promoter hypermethylation of tumor suppressor genes in other cancers --- p.39 / Chapter 2.5.3 --- Microsatellite instability --- p.42 / Chapter Chapter 3 --- The objectives of study --- p.45 / Chapter Chapter 4 --- Materials and Methods --- p.46 / Chapter 4.1 --- Samples --- p.46 / Chapter 4.1.1 --- Formalin fixed paraffin embedded tissues --- p.46 / Chapter 4.1.2 --- Cell lines --- p.46 / Chapter 4.2 --- Histological grading and staging of samples --- p.47 / Chapter 4.3 --- Microdissection on tissue sections --- p.47 / Chapter 4.4 --- Extraction of nucleic acid / Chapter 4.4.1 --- Extraction of DNA from paraffin-embedded tissues --- p.48 / Chapter 4.4.2 --- Extraction of DNA from cell lines --- p.49 / Chapter 4.5 --- DNA methylation analysis --- p.49 / Chapter 4.5.1 --- Overview of Methylation-Specific PCR (MSP) --- p.49 / Chapter 4.5.2 --- Bisulfite modification of DNA --- p.50 / Chapter 4.5.3 --- Methylation specific PCR (MSP) --- p.51 / Chapter 4.6 --- Microsatellite Analysis --- p.53 / Chapter 4.7 --- Statistical analysis --- p.56 / Chapter Chapter 5 --- Results / Chapter 5.1 --- Clinical-pathological features of endometrioid adenocarcinoma --- p.57 / Chapter 5.2 --- Promoter hypermethylation in endometrial cancer --- p.57 / Chapter 5.3 --- Microsatellite status (MSI) analysis --- p.65 / Chapter Chapter 6 --- Discussion / Chapter 6.1 --- Promoter hypermethylation in endometrial cancer --- p.71 / Chapter 6.1.1 --- Concurrent hypermethylation of multiple genesin endometrioid adenocarcinoma and its precursor lesions --- p.72 / Chapter 6.1.1.1 --- Promoter hypermethylation of E-cad --- p.73 / Chapter 6.1.1.2 --- Promoter hypermethylation of APC --- p.73 / Chapter 6.1.1.3 --- Promoter hypermethylation of MGMT --- p.74 / Chapter 6.1.1.4 --- Promoter hypermethylation of RASSF1A --- p.75 / Chapter 6.1.1.5 --- Promoter hypermethylation of hMLH-1 --- p.76 / Chapter 6.1.1.6 --- Promoter hypermethylation in ECA coexisting with hyperplasia and not coexisting with hyperplasia --- p.77 / Chapter 6.1.2 --- Promoter hypermethylation in SCA --- p.77 / Chapter 6.2 --- Microsatellite status analysis --- p.78 / Chapter 6.2.1 --- MSI in endometrial cancer --- p.78 / Chapter 6.2.2 --- MSI and concurrent promoter hypermethylation --- p.79 / Chapter 6.2.3 --- MSI and promoter hypermethylation of hMLH-1 --- p.80 / Chapter Chapter 7 --- Conclusion --- p.81 / Further studies --- p.82 / References --- p.83

Lee Mei Lan May. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 48-56). / Abstracts in English and Chinese. / ABSTRACT --- p.I / 中文摘要 --- p.III / ACKNOWLEDGEMENTS --- p.IV / PUBLICATION --- p.V / LIST OF FIGURES --- p.VI / LIST OF TABLES --- p.VII / ABBREVIATION --- p.VIII / CONTENTS --- p.IX / Chapter CHAPTER ONE: --- INTRODUCTION AND LITERATURE / Chapter 1.1 --- THYROID CANCER AND ITS EPIDEMIOLOGY --- p.1 / Chapter 1.1.1 --- Histology --- p.1 / Chapter 1.1.2 --- Gender comparison --- p.3 / Chapter 1.1.3 --- Female hormone 一 risk factor --- p.6 / Chapter 1.2 --- BIOLOGICAL BACKGROUND OF HORMONE´-´ؤؤ --- p.7 / Chapter 1.2.1 --- Estrogen --- p.8 / Chapter 1.2.2 --- Estrogen antagonist --- p.9 / Chapter 1.2.3 --- Estrogen receptors --- p.10 / Chapter 1.2.4 --- Estrogen receptor and thyroid cancer --- p.12 / Chapter 1.3 --- THE ROLE OF APOPTOSIS --- p.13 / Chapter 1.3.1 --- Bcl-family protein --- p.13 / Chapter 1.3.2 --- Bcl-family protein and cancer --- p.14 / Chapter 1.3.3 --- Estrogen and Bcl-family protein --- p.15 / Chapter 1.4 --- Objectives --- p.16 / Chapter CHAPTER TWO: --- GENERAL MATERIALS AND METHODS / Chapter 2.1 --- MATERIALS --- p.17 / Chapter 2.1.1 --- Culture media and treatment reagents --- p.17 / Chapter 2.1.2 --- Reagents for Western blot assay --- p.18 / Chapter 2.1.3 --- Antibodies --- p.19 / Chapter 2.1.4 --- Materials for RT-PCR --- p.19 / Chapter 2.1.5 --- Kits --- p.20 / Chapter 2.1.6 --- Instrumentations --- p.20 / Chapter 2.2 --- Methods --- p.21 / Chapter 2.2.1 --- Cell culture and treatment --- p.21 / Chapter 2.2.2 --- MTT assay --- p.22 / Chapter 2.2.3 --- Western blot analysis --- p.23 / Chapter 2.2.3.1 --- Protein extraction --- p.23 / Chapter 2.2.3.2 --- SDS-PAGE and protein transfer --- p.23 / Chapter 2.2.3.3 --- Immunoblotting analysis --- p.24 / Chapter 2.2.4 --- RNA extraction and RT-PCR --- p.25 / Chapter 2.2.4.1 --- RNA extraction --- p.25 / Chapter 2.2.4.2 --- cDNA synthesis --- p.26 / Chapter 2.2.4.3 --- Polymerase Chain Reaction (PCR) --- p.27 / Chapter 2.2.5 --- Statistical analysis --- p.28 / Chapter CHAPTER THREE: --- RESULTS / Chapter 3.1 --- Effect of E2 and tamoxifen on proliferation --- p.29 / Chapter 3.2 --- Comparison of effects of E and testosterone on Proliferation --- p.31 / Chapter 3.3 --- Differential Bcl-xL expression in response to E2 and testosterone stimulation --- p.33 / Chapter 3.4 --- Expression of ERα and ERβ in response to E2 stimulation --- p.35 / Chapter 3.5 --- Bcl-xL and Bax protein expression in response to E2 stimulation --- p.36 / Chapter 3.6 --- Expression of Bcl-xL and Bax mRNA in response to E2 stimulation --- p.39 / Chapter CHAPTER FOUR: --- DISCUSSION --- p.41 / Chapter CHAPTER FIVE: --- CONCLUSION --- p.47 / REFERENCES --- p.48