CSCs, caracterizadas pela alta atividade da ALDH1 e expressão (ou não) de marcadores de células-tronco embrionárias OCT-4, NANOG, SOX2, KLF4, LIN2, estão presentes em tumores de mama HER2+. Mamosferas são estruturas celulares esferóides in vitro enriquecidas em CSCs. Neste trabalho, buscou-se um melhor entendimento do papel dos miRNAs -221, -222 e -4728-3p na biologia das CSCs. Foi observado alto percentual de células ALDH1+ (citometria) em mamosferas das linhagens MCF-7, SKBR3 e BT-474; alta expressão (qRT-PCR) de miR-221 e -222 em mamosferas da linhagem MCF-7 e de tumores de pacientes, além de alta expressão de HER2 em mamosferas das linhagens MCF-7 e SKBR3. A MFE de células MCF-7 mostrou-se aumentada após a indução (com lentivetores) da superexpressão dos miRNAs -221 e -222. A resistência ao quimioterápico paclitaxel estava aumentada em mamosferas que superexpressavam o miRNA-222. A superexpressão de miR-4728-3p, localizado em um intron de HER2, levou a um aumento das subpopulações ALDH1+ em duas linhagens celulares reforçando seu envolvimento com a biologia de CSCs. / CSCs, characterized by high activity of ALDH1 and high (or not) expression of embryonic stem cell markers OCT-4, NANOG, SOX2, KLF4, LIN28 are present in tumors with HER2 amplification. Mammospheres are spheroid cell structures in vitro enriched by CSCs. In this study, we aim to contribute to the better understanding the role of miRNAs -221, 222, 4728-3p in the the biology of CSCs. We observed high percentage of ALDH+ cells (cytometry) in mammospheres of MCF-7, SKBR3 and BT-474 cell lines; high expression (qRT-PCR) of miR-221 and -222 in mammospheres of MCF-7 cell line and cells derived from patients, moreover HER2 is upregulated in mammospheres from MCF-7 and SKBR3 cell lines. MFE is higher in MCF-7 cell line after induction (with lentivectors) of miR-221 and -222 expression. The resistance to paclitaxel was increased in mammopheres that were overexpressing miRNA-222. Overexpression of miR-4728-3p, located in an intron of HER2 gene, induced the increment of ALDH1+ subpopulations in two cell lines, reinforcing its role in the biology of CSCs.

Um exame da literatura nacional e internacional evidencia uma quantidade considerável de publicações científicas dedicadas às repercussões psicológicas do câncer de mama e de seu tratamento. No entanto, a abordagem das questões relacionadas ao apoio social e familiar, na perspectiva da mulher acometida, é ainda pouco investigada. Para fundamentar as ações de saúde e de cuidado integralizado em saúde, torna-se necessária a realização de estudos que busquem compreender de que modo o apoio social e familiar pode contribuir para minimizar as repercussões do diagnóstico e do tratamento da neoplasia mamária para a paciente. Estudos têm enfocado a perspectiva do familiar, mas são escassos os trabalhos que se propõem a investigar a visão da própria mulher em tratamento. Nesse sentido, o presente estudo tem por objetivo compreender como a mulher que se encontra em processo de reabilitação do câncer de mama vivencia o cuidado que lhe é oferecido pelas redes sociais de apoio, particularmente em relação ao apoio social e familiar percebido. Participaram do estudo sete colaboradoras com idades entre 39 e 73 anos de idade, submetidas à mastectomia no período de julho de 2004 a junho de 2008, assistidas por um núcleo de reabilitação de mulheres mastectomizadas da Escola de Enfermagem de Ribeirão Preto. Para alcançar o objetivo proposto, utilizou-se a Fenomenologia enquanto referencial teórico-metodológico de pesquisa qualitativa. Dentro desse enfoque, a coleta de dados foi constituída por duas etapas: na primeira, foram realizadas entrevistas individuais e audiogravadas, mediante o consentimento das colaboradoras. A primeira etapa da coleta foi desenvolvida a partir de uma questão norteadora: Eu gostaria que você me contasse como tem vivenciado o apoio familiar desde que soube que estava com câncer de mama e o que isso significa para você. Na segunda etapa da coleta de dados, foram elaborados, conjuntamente com as participantes, o genograma e o ecomapa, considerando-se dois momentos distintos: antes e após o diagnóstico do câncer de mama; também foi aplicado o Critério de Classificação Econômica Brasil 2008. O material proveniente desses instrumentos permitiu a caracterização das participantes do estudo e de suas redes familiares, além da identificação do apoio recebido. As entrevistas foram transcritas na íntegra e analisadas juntamente com os demais dados, segundo o referencial metodológico adotado, por meio da descrição, redução e interpretação fenomenológicas. Os resultados mostraram que a rede de apoio social e familiar da paciente foi consideravelmente ampliada e diversificada após o diagnóstico do câncer de mama, o que não necessariamente significou para a mulher em tratamento um aumento de recursos, uma vez que nem sempre ela estava apta a dar conta de atribuir significados positivos aos cuidados que recebe para seu bem-estar psicossocial. Espera-se que o conhecimento resultante deste estudo possa auxiliar os profissionais a capacitar o processo de significação de mulheres com câncer de mama a respeito do apoio social e familiar que recebem, ampliando as possibilidades de compreensão dos cuidados que lhes são ofertados. / A review of national and international literature evidences a considerable amount of scientific studies addressing psychological repercussions of breast cancer and its treatment. However, issues related to social and family support from the perspective of affected women is still little investigated. Studies aimed to understand how social and family support can minimize the repercussions of a breast cancer diagnosis and treatment on patients are needed to ground health actions and integral health care delivery. Studies emphasized the perspective of family members but there are few studies investigating the view of women under treatment. From this perspective, this study aims to understand how women in breast cancer rehabilitation process experiences care provided by social support networks, specifically in relation to perceived social and family support. Seven women aged between 39 and 73 years submitted to mastectomy between July 2004 and June 2008 and attending a center for mastectomized women rehabilitation at the University of São Paulo at Ribeirão Preto, College of Nursing participated in the study. This qualitative study used Phenomenology as the theoreticalmethodological framework. Data collection was carried out in two stages. The first stage included individual interviews guided by the question Would you tell me how you have experienced family support since you have learned you had breast cancer and what it means to you? Interviews were audio recorded after the participants\' consent. In the second stage a genogram and ecomap were developed jointly with the participants considering two distinct points in time: before and after the breast cancer diagnosis. The Brazilian Economic Classification Criteria was also applied. Data obtained from these instruments permitted to characterize the study\'s participants and their family networks in addition to received support. Interviews were transcribed verbatim and analyzed jointly with the remaining data according to the adopted methodological reference through phenomenological description, reduction and interpretation. The results show that the patients\' social and family support networks were considerable enlarged and diversified after the breast cancer diagnosis, which however, did not necessarily mean they had increased resources since the women under treatment were not always able to attribute positive meanings to the care received for their psychosocial wellbeing. The knowledge acquired with this study is expected to help professionals to improve the signification process of women with breast cancer concerning the social and family support they receive, broadening understanding of care delivery.

O interesse da comunidade científica pelos aspectos psicológicos do câncer de mama aumentou consideravelmente nas duas últimas décadas. Diversas pesquisas com esse enfoque sugerem que a personalidade das pacientes pode influenciar o curso da doença, mas poucas contemplam especificamente essa questão. O presente estudo teve como objetivo comparar aspectos dinâmicos e elementos estruturais de personalidade em dois grupos distintos, em termos do curso da doença, de mulheres acometidas por câncer de mama. A amostra foi constituída por um grupo de mulheres em remissão (n=8), denominado Grupo 1, e um grupo de mulheres em recidiva (n=8), denominado Grupo 2. Todas as participantes eram vinculadas a uma entidade assistencial, cumpriram o tratamento médico preconizado e não apresentavam antecedentes psiquiátricos. A coleta de dados foi conduzida mediante o emprego de entrevista psicológica e do Teste de Apercepção Temática (TAT). Os dados obtidos foram inicialmente submetidos a análises qualitativas, descritivas e exploratórias e, posteriormente, organizados em estudos de caso. Além disso, foram interpretados sob a ótica da sicossomática psicanalítica a partir de uma articulação entre as proposições teóricas de Marty e McDougall. Considerando-se o estado da arte na atualidade, trata-se de um desenho metodológico inovador. Os resultados revelam que, na maioria das articipantes do Grupo 1, a integração lógica, a capacidade adaptativa e o funcionamento defensivo são apropriados, mas o controle dos impulsos é restritivo. As imagos parentais têm valência essencialmente positiva e a dinâmica das relações interpessoais varia da dominância à submissão. Conflitos associados à oscilação entre desejos de passividade e rigidez ou à insustentabilidade de ideais narcísicos se mostraram típicos e o contato com a própria subjetividade pareceu penoso, mas viável. O Grupo 2 diferenciou-se porque, na maioria das participantes, a capacidade adaptativa é comprometida por uma veemente dificuldade na utilização da realidade interna como referência para a organização da realidade externa. O funcionamento defensivo se sustenta à custa do apelo descontextualizado a operações psíquicas que sugerem uma estruturação egóica pouco consistente. O controle dos impulsos é restritivo e as imagos parentais têm valência predominantemente negativa. Além disso, um apego excessivo ao factual decorrente do apagamento da dinâmica mental conduz à conformidade social dos afetos, à contenção da atividade imaginativa ou até mesmo à estase libidinal. É possível afirmar, portanto, que o pensamento operatório e a desafetação são as principais características da personalidade das mulheres em situação de recidiva avaliadas. Conseqüentemente, pode-se situar o retorno do câncer na trama do trauma ensejado pelos desdobramentos do diagnóstico e do tratamento do tumor primário, uma vez que tais características promovem a substituição da simbolização pela reação biológica quando da ocorrência de eventos potencialmente desestruturantes. O presente estudo, assim, favorece uma compreensão inicial dos mecanismos psicodinâmicos por meio dos quais a personalidade das pacientes é capaz de influenciar o curso do câncer de mama. / The scientific community has become increasingly interested in the psychological aspects of breast cancer over the last two decades. Several studies on this issue suggest that patients personality can affect the course of the disease, but only a few address this issue specifically. The purpose of the present study was to compare dynamic aspects and structural elements of personality in two distinct groups of women with breast cancer, in terms of the course of the disease. The sample consisted of a group of women in remission (n=8), Group 1, and another of women with recurrence (n=8), Group 2. Every participant was registered in a communitarian entity, followed the recommended medical treatment, and presented no psychiatric history. Data collection was performed through psychological interviews and administrating the Thematic Apperception Test (TAT). The obtained data were initially submitted to qualitative, descriptive, and exploratory analyses, and, later, organized in case studies. In addition, the data were interpreted under a psychoanalytic psychosomatic view based on the articulation between the theoretical propositions of Marty and McDougall. Considering the current state-of-the-art, this study has an innovative methodological design. The results show that most participants in Group 1 have an appropriate logical integration, adjustment capacity, and defensive functioning, but their impulse control is restrictive. The parental imago have essentially positive associations and the dynamics of the interpersonal relationships ranges from dominance to submission. The conflicts were associated with the oscillation between desires of passivity and rigidity or to the insustainability of narcissistic ideals were typical and the contact with ones own subjectivity seemed pitiful, but feasible. Group 2 was different because most participants adjustment capacity was compromised by a great difficulty to use their internal reality as a reference to organize their external reality. Their defensive function is sustained by the decontextualized appeal to psychological operations that suggest a poorly consistent ego structure. Impulse control is restrictive and the parental imago had a predominantly negative association. In addition, an excessive attachment to the reality due to erasing the mental dynamics led to social conformity of affect, to the contention of the imaginative activity or even to libidinal stasis. Therefore, it can be stated that operational thinking and disaffectation are the main personality characteristics of women in recurrence, evaluated in this study. Consequently, cancer recurrence can be inserted in the plot of trauma caused by the outcomes of diagnosis and treatment of the primary tumor, since these characteristics promote the replacement of symbolization by the biological reaction when potentially destructuring events occur. The present study, thus, favors the initial comprehension of the psychodynamic mechanisms through which the patients personality is capable of influence the course of breast cancer.

O INKKI é um peptídeo catiônico anfipático derivado da ?-caseina bovina, análogo ao Mastoparan, isolado do veneno da vespa Vespula lewisi. Diferentes estudos mostraram sua atividade antiproliferativa e antitumoral in vitro com citotoxidade seletiva em linhagens de células de melanoma B16F10 em relação às células normais. Em estudos In vivo, INKKI demonstrou potente ação estimuladora da atividade fagocitária de macrófagos peritoneais. Quando aplicado diretamente em tumores, reduziu a massa tumoral, diminuindo o tempo de progressão do tumor e a formação de metástases, aumentando a probabilidade de sobrevida. Neste trabalho foram investigados os efeitos in vitro do peptídeo INKKI em linhagens de células de adenocarcinoma de mama e em células normais. As linhagens de células tumorais de mama utilizadas foram MDA-231, MCF-7 e T47D humanas, mama murina de Ehrlich e, como células normais, os fibroblastos e as células endoteliais. Foram avaliadas a atividade citotóxica, a viabilidade celular, a alteração do potencial elétrico mitocondrial, os efeitos na progressão e parada do ciclo celular e a investigação da morte por apoptose. Os resultados revelaram que o peptídeo INKKI apresenta citotoxidade seletiva dose-dependente nas células tumorais, modulando negativamente o potencial elétrico mitocondrial. O índice de proliferação celular das células tratadas diminuiu, com a parada do ciclo celular na fase G0/G1. O peptídeo induziu a morte celular por apoptose, a qual demonstrou ocorrer dependente da via mitocondrial e de forma caspaseindependente. O peptídeo INKKI é um potente modulador das atividades antiproliferativa e antitumoral da linhagem de células de adenocarcinoma de mama humana MCF-7 / The INKKI is derived from the bovine hidrolisis of ?-casein, a cationic and amphipathic peptide, analogous to mastoparan peptide isolated from the venom of the wasp Vespula lewisi. In in vitro studies, INKKI showed selective cytotoxicity in melanoma tumor cell B16F10 in relation to normal cells. In the in vivo tests, INKKI peptide has been shown to be a potent stimulator of the phagocytic activity of treated macrophages and, when applied directly into tumors, reduced tumor mass, decreasing the time progression and metastasis formation, increasing the probability of survival. This work investigated the in vitro effects of tumor peptide INKKI in cell line of human breast adenocarcinoma MCF-7. The tumor cells used were lineages of human breast tumor cells MCF7, MDA-231 and T47D and murine tumor cells (Ehrlich). The normal cells used were fibroblasts and endothelial cells. The study evaluated the cytotoxic activity, cell viability, change of mitochondrial electrical potential, effects on the progression and arrest of the cell cycle and investigation of induced death by apoptosis. The results revealed that peptide presents selective dose-dependent cytotoxicity in tumor cells by modulating negatively the mitochondrial electric potential. The index of cell proliferation of cells treated declined, with the induction of cell cycle arrest at phase G0/G1. The peptide-induced cell death by apoptosis was demonstrated to occur via mitochondrial pathway and in a caspase-independent manner. The INKKI peptide is a potent modulator of antiproliferative and antitumor activities of the breast adenocarcinoma cell line

A dor e o linfedema são as principais sequelas que acometem as pacientes com câncer de mama após o tratamento dessa doença, e essas sequelas acarretam alterações físicas e psicológicas que podem modificar a qualidade de vida relacionada à saúde (QVRS) dessas pacientes. Este estudo teve como objetivo verificar a presença da dor, do linfedema e da QVRS nas pacientes com câncer de mama. Foram realizadas avaliações da presença da dor com a EVA, da presença do linfedema com a perimetria e da QVRS com o EORTC QLQ C30. A amostra constituiu-se de 181 pacientes com câncer de mama que faziam acompanhamento médico da doença no Ambulatório de Mastologia do HCFMRP-USP e no Ambulatório de Fisioterapia da Fundação Pio XII - Hospital do Câncer de Barretos. A média da idade das pacientes variou de 52,81 (±11,68) a 54,09 (±11,39); a maioria das pacientes era casada, com baixo nível de escolaridade e desempenhavam atividade doméstica. A mama esquerda foi a mais acometida, o carcinoma ductal infiltrante foi o predominante e a mastectomia foi a técnica cirúrgica mais utilizada nas pacientes deste estudo. Constatou-se que 78 pacientes referiram sentir dor no membro superior homolateral à intervenção cirúrgica, em 52 pacientes constatou a presença do linfedema. A QVRS foi considerada como boa pelas pacientes. / The pain and lymphedema are the major consequences that affect patients with breast cancer after treatment of this disease, and these consequences cause physical and psychological changes that may alter the quality of life related to health of these patients (HRQOL). This study aimed to verify the presence of pain, lymphedema and HRQOL in patients with breast cancer. The presence of pain evaluated using visual anologue scale (VAS), perimetry was used to evaluate presence of lymphedema and HRQOL was measure with EORTC QLQ C30. The sample consisted of 181 patients with breast cancer who were monitoring the disease at the Mastology Outpatient of the Hospital das Clínicas of the University of São Paulo at Ribeirão Preto Medical School (HCRPFMRP/USP) and the Physiotherapy Outpatient of the Pius XII Foundation - Cancer Hospital of Barretos. The mean age of the patients ranged from 52,81 (±11,68) to 54,09 (±11,39). Most patients were married, with a low educational level and housewifes. The left breast was the most affected, the infiltrating ductal carcinoma was the most prevalent and mastectomy surgical technique was used in most patients in this study. 78 patients reported pain in the homolateral upper limb subsequent to surgery, in 52 patients had lymphedema. HRQOL was considered good for patients.

陳津利. / "2000年7月" / 論文 (哲學碩士)--香港中文大學, 2000. / 參考文獻 (leaves 138-151) / 附中英文摘要. / "2000 nian 7 yue" / Chen Jinli. / Lun wen (zhe xue shuo shi)--Xianggang Zhong wen da xue, 2000. / Can kao wen xian (leaves 138-151) / Fu Zhong Ying wen zhai yao. / Chapter 第一章 --- 緒論 --- p.1 / Chapter 第一節 --- 中國的乳腺癌患者 --- p.1 / Chapter 第二節 --- 中國內地的癌症自助團體 --- p.3 / Chapter 第三節 --- 硏究意義和主題 --- p.5 / Chapter 一、 --- 硏究意義 --- p.5 / Chapter 二、 --- 硏究主題 --- p.8 / Chapter 第二章 --- 文獻回顧 --- p.9 / Chapter 第一節 --- 乳腺癌和乳腺癌患者 --- p.9 / Chapter 一、 --- 乳腺癌和乳腺癌患者的定義 --- p.9 / Chapter 二、 --- 患者瀕死的心理反應 --- p.10 / Chapter 三、 --- 患者面臨的實際問題和情感需求 --- p.15 / Chapter 第二節 --- 支持的槪念和取向 --- p.21 / Chapter 一、 --- 社會支持的槪念 --- p.21 / Chapter 二、 --- 支持的類型 --- p.23 / Chapter 三、 --- 社會支持的功能 --- p.27 / Chapter 四、 --- 社會支持的測量和指標 --- p.31 / Chapter 第三節 --- 自助團體的定義及功能 --- p.36 / Chapter 一、 --- 自助團體的定義 --- p.36 / Chapter 二、 --- 自助團體作爲社會支持系統的組成部分 --- p.37 / Chapter 第三章 --- 硏究架構 --- p.46 / Chapter 第一節 --- 基本假設 --- p.46 / Chapter 第二節 --- 研究變量名詞的操作性定義 --- p.48 / Chapter 一、 --- 自助團體行爲認知 --- p.48 / Chapter 二、 --- 感受的情感支持 --- p.50 / Chapter 三、 --- 個人特徵 --- p.50 / Chapter 第四章 --- 硏究方法 --- p.51 / Chapter 第一節 --- 研究法的選擇 --- p.51 / Chapter 一、 --- 應用三角測量的思考 --- p.51 / Chapter 二、 --- 量化爲主、質性爲輔的主輔設計 --- p.51 / Chapter 三、 --- 同步三角測量法的采用 --- p.53 / Chapter 第二節 --- 問卷調查硏究設計 --- p.54 / Chapter 一、 --- 樣本策略_ --- p.54 / Chapter 二、 --- 測量工具 --- p.56 / Chapter 三、 --- 資料搜集方法 --- p.59 / Chapter 四、 --- 效度和信度 --- p.59 / Chapter 五、 --- 問卷調查的資料處理及數据分析 --- p.62 / Chapter 第三節 --- 質性硏究方案 --- p.63 / Chapter 一、 --- 硏究樣本及資料收集 --- p.63 / Chapter 二、 --- 資料分析 --- p.63 / Chapter 第四節 --- 硏究局限 --- p.65 / Chapter 第五章 --- 硏究結果 --- p.66 / Chapter 第一節 --- 中國乳腺癌患者的基本特徵 --- p.66 / Chapter 一、 --- 背景結構 --- p.66 / Chapter 二、 --- 身心狀況 --- p.72 / Chapter 三、 --- 病患情感特徵 --- p.77 / Chapter 第二節 --- 患者自助團體行爲認知 --- p.85 / Chapter 一、 --- 患者的自助團體活動參與程度(A1 ) --- p.85 / Chapter 二、 --- 患者的自助團體活動認知程度(A2 ) --- p.90 / Chapter 三、 --- 患者對自助團體的主觀認同(A3) --- p.91 / Chapter 四、 --- 個人基本特徵的作用 --- p.96 / Chapter 第三節 --- 患者自助團體中的情感支持感受(B) --- p.100 / Chapter 一、 --- 患者在自助團體中的情感支持感受 --- p.100 / Chapter 二、 --- 個人特徵對情感支持感受影響 --- p.108 / Chapter 第四節 --- 假設驗証：自助團體行爲認知與情感支持感受的 相關關係 --- p.114 / Chapter 一、 --- 活動參與程度(A1)與情感支持感受的相關關係 --- p.115 / Chapter 二、 --- 活動認知程度(A2)與情感支持感受的相關關係 --- p.119 / Chapter 三、 --- 主觀認同(A3)與情感支持感受的相關關係 --- p.120 / Chapter 四、 --- 個人基本特徵對行爲認知(A)與情感支持感受 關係的影響 --- p.122 / Chapter 五、 --- 小結 --- p.122 / Chapter 第六章 --- 結論、檢討及建議 --- p.123 / Chapter 第一節 --- 硏究結果討論 --- p.123 / Chapter 一、 --- 自助團體情感支持硏究結果與西方 研究的异同及討論 --- p.123 / Chapter 二、 --- 社會支持理論和槪念的适用性討論 --- p.127 / Chapter 第二節 --- 待5幵究或澄淸的議題 --- p.129 / Chapter 一、 --- 患者的情感問題： 乳腺癌是否破坏了患者的婚姻質量？ --- p.129 / Chapter 二、 --- 家庭的支持功能問題： 什么是對患者的“愛´ح ？ --- p.131 / Chapter 三、 --- 來自“大鍋飯´ح的資源： 制度安排帶給乳腺癌患者什么？ --- p.131 / Chapter 四、 --- 未接受乳房再造術： 經濟的？醫療的？制度的？觀念的？ --- p.132 / Chapter 五、 --- 關於自助團體的几點發現 --- p.132 / Chapter 第三節 --- 建議 --- p.134 / Chapter 一、 --- 政府部門與自助團體建立伙伴關係 --- p.134 / Chapter 二、 --- 加強對患者家庭的輔導，引進境外社會工 作理念和實務，開展針對性情感治療 --- p.134 / Chapter 三、 --- 營造新女性文化，關注自身生存質量 --- p.135 / Chapter 四、 --- 未來的硏究 --- p.136 / 參考書目 --- p.138 / 附錄 / 附錄一統計結果列表 --- p.152 / 附錄二 量表信度列表 --- p.173 / 附錄三調查問卷 --- p.177 / 附錄四訪談指引 --- p.185

by Chow Ka Yee. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 203-221). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / Abstract in Chinese --- p.iv / List of Abbreviations --- p.vi / Table of Contents --- p.xi / List of Figures --- p.xviii / List of Tables --- p.xxii / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- The Characteristics of Arsenic Trioxide (AS2O3) --- p.2 / Chapter 1.2 --- The Therapeutic Applications of Arsenic Trioxide (As203) --- p.5 / Chapter 1.3 --- Acute Promyelocytic Leukemia (APL) --- p.6 / Chapter 1.3.1 --- Pathologies of APL --- p.7 / Chapter 1.3.2 --- All Trans Retinoic Acid (ATRA) Treatment of APL Patients --- p.7 / Chapter 1.3.3 --- Clinical Trials of Arsenic Trioxide (As203) on APL Patients --- p.9 / Chapter 1.3.4 --- In Vitro and In Vivo Studies of Arsenic Trioxide (As203) in the Treatment of APL --- p.10 / Chapter 1.3.4.1 --- Induction of Apoptosis --- p.11 / Chapter 1.3.4.2 --- Induction of Cell Differentiation --- p.11 / Chapter 1.3.5 --- General Toxicity and Side Effects of Arsenic Trioxide (AS2O3) on APL Patients --- p.12 / Chapter 1.4 --- Effects of Arsenic Trioxide (As203) on Other Primary Cancer Cells and Cancer Cell Lines --- p.12 / Chapter 1.5 --- Epidemiology of Breast Cancer --- p.14 / Chapter 1.6 --- Classification of Breast Cancer --- p.17 / Chapter 1.7 --- Etiology of Breast Cancer --- p.17 / Chapter 1.8 --- Hormones and Breast Cancer --- p.18 / Chapter 1.9 --- Estrogen Receptors (ER) --- p.20 / Chapter 1.9.1 --- Structures of Estrogen Receptors (ER) --- p.21 / Chapter 1.9.2 --- Estrogen Receptors (ER) Mediated Signaling Pathway --- p.22 / Chapter 1.9.2.1 --- Ligand Dependent Pathway --- p.22 / Chapter 1.9.2.2 --- Ligand Independent Pathway --- p.22 / Chapter 1.9.2.3 --- Estrogen Response Element (ERE)-Independent Pathway --- p.23 / Chapter 1.9.2.4 --- Non-Genomic Pathway --- p.23 / Chapter 1.9.3 --- Estrogen Receptors (ER) Regulated Gene Expression --- p.25 / Chapter 1.10 --- Current Therapy of Breast Cancer --- p.26 / Chapter 1.10.1 --- Hormonal Therapy (Anti-Estrogenicity) --- p.26 / Chapter 1.10.1.1 --- Tamoxifen --- p.26 / Chapter 1.10.1.2 --- Other Pure Anti-Estrogens --- p.28 / Chapter 1.10.2 --- Regulation of Estrogen Receptors (ER) and Transcription Coregulators --- p.29 / Chapter 1.10.3 --- Apoptosis Induction --- p.29 / Chapter 1.11 --- Aims of Study --- p.30 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.32 / Chapter 2.1 --- Materials --- p.33 / Chapter 2.1.1 --- Cell Lines and Culture Media --- p.33 / Chapter 2.1.1.1 --- Cell Lines --- p.33 / Chapter 2.1.1.2 --- Culture Media --- p.34 / Chapter 2.1.2 --- Chemicals --- p.35 / Chapter 2.1.3 --- Reagents and Buffers --- p.36 / Chapter 2.1.3.1 --- Reagents for MTT Assay --- p.36 / Chapter 2.1.3.2 --- Reagents for [methyl-3H] Thymidine Incorporation into DNA --- p.37 / Chapter 2.1.3.3 --- Reagents for Trypan Blue Exclusion Assay --- p.37 / Chapter 2.1.3.4 --- Reagents and Buffers for Western Blot Analysis --- p.37 / Chapter 2.1.3.5 --- Reagents and Buffers for Flow Cytometry --- p.40 / Chapter 2.1.3.6 --- Reagents and Buffers Reverse Transcription Polymerase Chain Reaction (RT-PCR) --- p.40 / Chapter 2.1.3.7 --- Reagents for Transfection and Luciferase Reporter Assay --- p.41 / Chapter 2.1.3.8 --- Reagents and Buffers for In Vivo Studies --- p.42 / Chapter 2.2 --- Methods --- p.42 / Chapter 2.2.1 --- In Vitro Studies --- p.42 / Chapter 2.2.1.1 --- Cell Treatment --- p.42 / Chapter 2.2.1.2 --- Drug Preparation --- p.43 / Chapter 2.2.1.3 --- MTT Assay --- p.43 / Chapter 2.2.14 --- Trypan Blue Exclusion Assay --- p.44 / Chapter 2.2.1.5 --- [methyl-3H] Thymidine Incorporation into DNA --- p.45 / Chapter 2.2.1.6 --- Detection of DNA Fragmentation --- p.45 / Chapter 2.2.1.7 --- ERα Competitive Binding Assay --- p.47 / Chapter 2.2.1.8 --- Cell Cycle Analysis by Flow Cytometry with Propidium Iodide (PI) Staining --- p.48 / Chapter 2.2.1.9 --- Cell Cycle Analysis by Flow Cytometry with Annexin V-PI Staining --- p.48 / Chapter 2.2.1.10 --- Cell Cycle Analysis by Flow Cytometry with JC-1 Staining --- p.49 / Chapter 2.2.1.11 --- Cell Cycle Analysis by Flow Cytometry with Hydroethidine (HE) Staining --- p.50 / Chapter 2.2.1.12 --- Western Blot Analysis of Proteins --- p.50 / Chapter 2.2.1.13 --- Assessment of the Transcriptional Activity of ERα --- p.55 / Chapter 3.2.1.14 --- Reverse Transcription Polymerase Chain Reaction (RT-PCR) --- p.57 / Chapter 2.2.2 --- In Vivo Studies --- p.61 / Chapter 2.2.2.1 --- Animal Models --- p.61 / Chapter 2.2.2.2 --- Treatment Schedules --- p.61 / Chapter 2.2.2.3 --- Sacrifice of Nude Mice --- p.61 / Chapter 2.2.2.4 --- Enzymatic Assays --- p.62 / Chapter 2.2.2.4.1 --- Aspartate Transaminase (AST) --- p.63 / Chapter 2.2.2.4.2 --- Alanine Transaminase (ALT) --- p.64 / Chapter 2.2.2.4.3 --- Creatine Kinase (CK) --- p.65 / Chapter 2.2.2.4.4 --- Lactate Dehydrogenase (LDH) --- p.66 / Chapter CHAPTER 3 --- "Effects of Arsenic Trioxide (As203) on Human Breast Adenocarcinoma Cell Line, MCF-7 Cell Line" --- p.68 / Chapter 3.1 --- Introduction --- p.69 / Chapter 3.2 --- Effect of As203 on Cell Survival of MCF-7 cells by MTT Assay --- p.70 / Chapter 3.3 --- Cytotoxicity of As203 on MCF-7 Cells by Trypan Blue Exclusion Assay --- p.72 / Chapter 3.4 --- Effect of As203 on DNA Synthesis and Cell Proliferation of MCF-7 cells by [methyl-3H] Thymidine Incorporation into DNA --- p.76 / Chapter 3.5 --- Comparison of Cytotoxicity of AS2O3 on MCF-7 Cells with that of Tamoxifen --- p.79 / Chapter 3.6 --- Summary --- p.82 / Chapter CHAPTER 4 --- Effects of Arsenic Trioxide (As203) on 17β Estradiol Stimulated MCF-7 cells --- p.83 / Chapter 4.1 --- Introduction --- p.84 / Chapter 4.2 --- Effect of 17β estradiol on Cell Viability of MCF-7 Cells by MTT Assay --- p.86 / Chapter 4.3 --- Effect of As203 and 17β Estradiol on Cell Survival of MCF-7 Cells by MTT Assay --- p.88 / Chapter 4.4 --- Cytotoxicity of As203 on 17β Estradiol Stimulated MCF-7 cells by Cell Number Counting with Hemacytometer --- p.92 / Chapter 4.5 --- Growth Inhibitory Effect of As203 on 17β Estradiol stimulated MCF-7 cells by [methyl-3H] Thymidine Incorporation into DNA --- p.94 / Chapter 4.6 --- "Effect of As203 on Cell Survival of Hormone Independent Breast Cancer Cell Line, MDA-MB-231 Cells" --- p.96 / Chapter 4.7 --- Summary --- p.100 / Chapter CHAPTER 5 --- Effects of Arsenic Trioxide (As203) on Normal Cells --- p.102 / Chapter 5.1 --- Introduction --- p.103 / Chapter 5.2 --- "Effect of As203 on Normal Human Fibroblast Cell Line, Hs68" --- p.104 / Chapter 5.3 --- Effects of As203 on the Normal Cells of Nude Mice --- p.106 / Chapter 5.3.1 --- Effect of AS2O3 on Aspartate Transaminase (AST) Activity of Nude Mice --- p.107 / Chapter 5.3.2 --- Effect of As203 on Alanine Transaminase (ALT) Activity of Nude Mice --- p.109 / Chapter 5.3.3 --- Effect of As203 on Creatine Kinase (CK) Activity of Nude Mice TABLE OF CONTENTS --- p.111 / Chapter 5.3.4 --- Effect of As203 on Lactate Dehydrogenase (LDH) Activity of Nude Mice --- p.113 / Chapter 5.4 --- Summary --- p.115 / Chapter CHAPTER 6 --- Action Mechanisms underlying the Survival Inhibitory Effects of Arsenic Trioxide (As203) on MCF-7 cells --- p.116 / Chapter 6.1 --- Introduction --- p.117 / Chapter 6.2 --- Detection of Apoptosis --- p.119 / Chapter 6.2.1 --- Detection of DNA Fragmentation --- p.119 / Chapter 6.2.2 --- Phosphatidylserine (PS) Externalization Detected by Flow Cytometry with Annexin V-PI Staining --- p.124 / Chapter 6.2.2.1 --- The Principle --- p.124 / Chapter 6.2.2.2 --- PS Externalization upon AS2O3 Treatment --- p.126 / Chapter 6.3 --- Analysis of Cell Cycle Distribution of MCF-7 Cells --- p.130 / Chapter 6.3.1 --- The Principle --- p.130 / Chapter 6.3.2 --- Regulation of Cell Cycle Distribution of MCF-7 Cells upon As2O3 Treatment --- p.131 / Chapter 6.4 --- The Action Mechanisms Underlying As203 Induced Apoptosis or Cell Cycle Arrest --- p.137 / Chapter 6.4.1 --- Effect of As203 on Mitochondrial Membrane Potential of MCF-7 Cells --- p.137 / Chapter 6.4.2 --- Regulation of Free Oxidative Species (ROS) Production in MCF-7 Cells upon AS2O3 Treatment --- p.140 / Chapter 6.4.2.1 --- Analysis of Superoxide Production in MCF-7 Cells upon AS2O3 Treatment by Flow Cytometry with Hydroethidine (HE) Staining --- p.140 / Chapter 6.4.2.2 --- Effect of As203 on Cell Survival of MCF-7 Cells Co-treated with N-Acteyl-L-Cysteine (NAC) by MTT Assay --- p.143 / Chapter 6.4.3 --- Regulation of Bcl-2 Protein Level in MCF-7 Cells upon As2O3 Treatment --- p.145 / Chapter 6.4.4 --- Regulation of p53 Protein Level in MCF-7 Cells upon AS2O3 Treatment --- p.147 / Chapter 6.5 --- Summary --- p.149 / Chapter CHAPTER 7 --- Effects of Arsenic Trioxide (As203) on Estrogen Receptor a (ERα) Mediated Signaling Pathway in MCF-7 cells --- p.150 / Chapter 7.1 --- Introduction --- p.151 / Chapter 7.2 --- Effect of As203 on Estrogen Binding to Estrogen Receptor a (ERα) by ERα Competitive Binding Assay --- p.152 / Chapter 7.3 --- Regulation of Estrogen Receptor a (ERα) mRNA Level upon As2O3 Treatment by RT-PCR --- p.156 / Chapter 7.4 --- Regulation of Estrogen Receptor a (ERα) Protein Level upon As2O3 Treatment --- p.159 / Chapter 7.5 --- Regulation of Estrogen Receptor a (ERα) Transcriptional Activity upon AS2O3 treatment --- p.161 / Chapter 7.6 --- "Regulation of Estrogen Target Gene, c-myc, Protein Level upon As2O3 Treatment" --- p.164 / Chapter 7.7 --- Effects of As203 on Cell Cycle Distribution of MCF-7 Cells under Estrogens Stimulation --- p.167 / Chapter 7.8 --- Summary --- p.173 / Chapter CHAPTER 8 --- Discussion --- p.174 / Chapter 8.1 --- The Anti-Tumor Effects of As203 on MCF-7 Cells --- p.175 / Chapter 8.2 --- Cytotoxicity of As203 on MCF-7 Cells --- p.175 / Chapter 8.2.1 --- Induction of Apoptosis in MCF-7 Cells upon As2〇3 Treatment --- p.176 / Chapter 8.2.2 --- Action Mechanisms Underlying the Induction of Apoptosis by As2〇3 --- p.178 / Chapter 8.3 --- Growth Inhibition of As203 on MCF-7 Cells --- p.182 / Chapter 8.3.1 --- Cell Cycle Regulation of MCF-7 Cells upon As203 Treatment --- p.182 / Chapter 8.4 --- Growth Inhibitory Effects of As203 on Estrogen Stimulated MCF-7 Cells --- p.186 / Chapter 8.4.1 --- Regulation of Estrogen Receptor a (ERα) Signaling Pathway in MCF-7 cells upon as2o3 Treatment --- p.188 / Chapter 8.5 --- Cross Talk of ERα Signaling Pathway and Apoptosis in Mediating the Anti-Tumor Effects of As203 on MCF-7 Cells --- p.195 / Chapter 8.6 --- Toxicity of AS2O3 towards Normal Tissues --- p.197 / Chapter CHAPTER 9 --- Conclusion and Future Perspectives --- p.200 / Chapter 9.1 --- Conclusion --- p.200 / Chapter 9.2 --- Future Perspectives --- p.202 / References --- p.203

Luo, Kewang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 278-305). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Yau, Ka Long. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 97-113). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts also in Chinese.

O projeto tem como objetivo a detecção e caracterização das células epiteliais circulantes de pacientes com carcinoma de mama no estádio III. Analisou-se a presença ou ausência destas células antes da realização de quimioterapia neoadjuvante. Para tanto, utilizou-se o sistema de enriquecimento de amostras de sangue periférico por meio da marcação intracelular imunomagnética direta, empregando-se microbeads conjugados com citoqueratina e posterior separação imunomagnética. A amostra final, com as células alvo detectadas, foi então quantificada por imunocitoquímica ou citometria de fluxo. Resultados: A reação de imunocitoquímica no sangue periférico foi negativa em todos os casos enquanto que na citometria de fluxo foi positiva em 22/23 casos analisados. Conclusão: A detecção e caracterização de células tumorais circulantes em pacientes com câncer de mama podem vir a se constituir em um novo fator prognóstico, porém, são ainda necessárias novas estratégias para melhor detectá-las. / The aim of this study was to detect and characterize carcinoma cells of patients with breast cancer with clinical stage III analyzing the presence or absence before chemotherapy by enrichment of peripheral blood with super paramagnetic microbeads, using direct Immunomagnetic labeling of intracellular 7/8 cytokeratin. For enrichment, magnetically labeled tumor cells are passed over a high-gradient magnetic positive selection column. Cytospin preparations are made after immunomagnetic enrichment and are examined immunocytochemically (ICC) using an anti-cytokeratin and quantified by flow cytometry (CMF). Results: No cytokeratin positive cells were detected in the 32 peripheral blood of patients of breast carcinoma by ICC and cytokeratin positive cells were detected in the 22 of 23 patients by CMF. Conclusion: These methods of detection and characterization breast carcinoma cells will become useful in the diagnosis and prognosis but is necessary optimization of existing methods.