Ping Zhang. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (p. 131-144). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Chin Wai. / Original paper published on European Jouranl of cancer (vol. 34, no. 7, p. 1015-1022) inserted. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1999. / Includes bibliographical references (leaves 70-77). / Abstracts in English and Chinese. / Dedication --- p.i / Vita --- p.ii / Acknowledgment --- p.iv / Abstract --- p.vii / Introduction --- p.1 / Hepatocellular carcinoma --- p.1 / Lung cancer --- p.3 / The role of suppressor gene PML in cancer --- p.5 / Principle of immunohistaining methods --- p.8 / Patients and methods --- p.21 / Patients and smaples --- p.21 / Slide preparing --- p.22 / Immunohistochemical staining --- p.23 / Cell culture --- p.30 / Determination of the population doubling times --- p.30 / Mtt assay --- p.35 / Results --- p.37 / "Altered expression of PML in normal liver, HCC and Secondary liver tumor" --- p.37 / Increased expression of PML in chronic hepatitis tissues --- p.38 / Differential expression of PML at the periphery and at the center of single-encapsulated lesion of HCC --- p.42 / Expression of PML in normal lung tissues --- p.43 / Suppression of PML expression in small cell lung cancer --- p.44 / Enhanced expression of PML in adenocarcinoma of the lung --- p.44 / Enhanced expression of PML in squamous cell carcinoma of the lung --- p.45 / Express of PML in metastatic lung cancer --- p.46 / Inverse correlation of the expression of PML and the proliferation marker Ki-67 in SCLC and SCC --- p.46 / Correlation of the expression of PML in macrophages with the macrophage-specific marker KP-1 --- p.47 / Expression of PML in Hela cells and Hela cells transfected with the gene --- p.48 / Altered morphology of the Hela-PML cell-clones --- p.49 / Altered growth rate in Hela-PML cells --- p.49 / Altered rate of cell-death in Hela-PML cells --- p.50 / Discussion --- p.51 / Further studies --- p.63 / References --- p.70 / Table --- p.78 / Figure legend --- p.81 / Appendix: Original paper published on European Journal of cancer --- p.106

Further studies were done to investigate the mechanism responsible for 1-BI-induced apoptosis in NCI-H460. It was found that 1-BI stimulated the expression of pro-apoptotic p53, Bax and cytosolic NF-kB p65 subunit but decreased pERK in NCI-H460 cells. The active forms of caspase 3 and caspase 9 were detected by Western blot, accompanied by an increase in caspase 3 activity. Reactive oxygen species (ROS) was highly generated at 24 hours after the treatment and the mitochondrial membrane potential was significantly decreased at 48 and 72 hours. The application of either N-acetyl cysteine (NAC) or glutathione (GSH) attenuated the cell growth inhibition caused by 1-BI. NCI-H460 cells pretreated with NAC showed a decrease in ROS production and p65 protein but an increase in pERK. / Taken together, these findings suggest that the inhibition of THXS suppresses lung cancer cell growth by promoting either G1 cell cycle arrest or apoptosis. The status of p53 is critical for both cell cycle arrest and apoptosis in 1-BI-mediated growth inhibition, which is evident by enhanced apoptosis detected in p53-transfected NCI-H23 and DMS 114 cells and G1 cell cycle in lung cancer cells treated with PFT-alpha. The 1-BI-induced growth-inhibitory pathway is associated with the generation of ROS, alteration of mitochondrial membrane potential, down-regulation of pERK and p65. / The result showed that THXS expressed in all of the three lung cancer cell lines (NCI-H23, DMS 114 and NCI-H460). The activity of THXS was also reflected by the presence of THXS metabolite thromobxane B2 (TXB2) in the cells, which was detected by ELISA. 1-Benzylimidazole (1-BI), a specific THXS inhibitor, suppressed the lung cancer cell proliferation measured by MTT assay. 1-BI treatment caused G1 phase arrest and enhanced the level of cyclin dependent kinase inhibitor p27 in a time-dependent manner in NCI-H23 and DMS 114 cells. It markedly increased DNA fragmentation in NCI-H460 cells. The findings suggest that 1-BI inhibits cell growth by arresting cell cycle and inducing cell death. Annexin V/PI staining revealed that the cell death induced by I-BI was mainly in the format of apoptosis. Further experiments showed that the I-BI-induced apoptosis could be enhanced by the introduction of p53 into NCI-H23 and DMS 114 cells, and such enhancement was associated with a decrease in mitochondrial membrane potential. This result suggests that the p53 may play a positive role in apoptosis induced by 1-BI through changing of the mitochondrial membrane potential. The role of p53 in I-BI-mediated apoptosis was further confirmed by the experiment of the p53 inhibition. Pifithrin-alpha hydrobromide (PFT-alpha), a p53 specific inhibitor, suppressed the 1-BI-induced p53 protein expression and increased G1 cell cycle arrest. / Thromboxane A2 (TXA2) is a potent arachidonate metabolite in the cyclooxygenase-2 (COX-2) pathway, which is produced by a member of cytochrome P450 (CYP) superfamily called thromboxane synthase (THXS). Recent studies have showed that thromboxane and THXS are associated with cancer cell migration, angiogenesis, tumor metastasis and cancer proliferation but there is limited information on their role in lung cancer development. This thesis is to test the hypothesis that inhibition of THXS could alter lung cancer cell growth. / Leung, Kin Chung. / Adviser: George G. Chen. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3319. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 130-144). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

In conclusion, our study has demonstrated that TGZ, a synthetic PPARgamma ligand, inhibits lung cancer cells growth through cell-cycle arrest, increased cell differentiation and induction of apoptosis. In this pathway, the activation of ERK by TGZ plays a central role in promoting apoptosis, which appears to be mediated via a mitochondria-related mechanism and functions in a PPARgamma-dependent manner. The interaction between PPARgamma and ERK may create an auto-regulatory and positive feedback loop to enhance the effect of ERK whereas the activation of Akt may generate a negative regulation to control the degree of apoptosis occurred in lung cancer cells. TGZ may counteract NNK function to inhibit lung cancer cell growth in the PPARgamma-dependent manner. / Lung cancer is the world's leading cause of cancer death. Currently there is not an acceptable adjuvant or palliative treatment modalities that have been conclusively shown to prolong survival in lung cancer. Therefore, translational research to improve outcomes with this disease is critical. Peroxisome proliferator-activated receptor-gamma (PPARgamma) is a member of the nuclear hormone receptor superfamily of ligand-activated transcription. PPARgamma ligands have been demonstrated to inhibit growth of cancer cells. The role of the PPARgamma in cell differentiation, cell cycle arrest and apoptosis has attracted increasing attention. Our study focused on the role of PPARgamma and its ligand troglitazone (TGZ) in the cell death of human lung cancer and the interaction between PPARgamma system and 4-(N-Methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a major tobacco-specific carcinogen. / The epidemic of lung cancer is directly attributable to cigarette. However, it is still not completely known the molecular pathway of cigarette smoking in the pathogenesis of lung cancer. Among the carcinogenoic chemicals of cigarette smoking, 4-(N-Methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is the most potent, which induces lung cancer in all animal species tested. Unlike PPARgamma ligands, NNK can promote cell proliferationa and growth. It is interesting to know whether PPARgamma ligands can inhibit the growth-promoting function of NNK. To address this question, we used NCI-H23 lung cancer cells as the model to study how TGZ influenced the function of NNK. Results showed that NNK stimulated cell proliferation, induced the DNA binding activity of nuclear factor-kappaB (NF-kappaB), down-regulated Bad expression, and up-regulated PPARgamma protein expressions. Inhibition of NF-kappaB nuclear translocation led to the suppression of NNK-mediated Bad expression, indicating that NNK may regulate Bad expression through the activation of NF-kappaB. TGZ significantly inhibited cell proliferation induced by NNK. Though TGZ did not affect nuclear factor-kappaB (NF-kappaB) activity, it up-regulated Bad expression. Taken together, TGZ can efficiently inhibit the proliferation of lung cancer cells induced by NNK via Bad- and PPARgamma- related pathways, which may not be directly relevant to the activity of NF-kappaB. / To elucidate the mechanism responsible for the effect of PPARgamma and TGZ on lung cancer cells, we further studied the PPARgamma molecular pathway in NCIH23 treated by TGZ. The result demonstrated that TGZ induced PPARgamma and ERK1/2 accumulation in the nucleus, where the co-localization of both proteins was found. It showed that the activation of ERK1/2 resulted in apoptosis via the mitochondrial pathway, reflecting by reduction of mitochondria membrane potential, change in Bcl-2 family members, release of cytochrome c into cytosol, and activation of caspase 9. Both PPARgamma siRNA and U0126, a specific inhibitor of ERK1/2, were able to block these effects of TGZ, suggesting that apoptosis induced by TGZ was PPARgamma- and ERK1/2-dependent. Inhibition of ERK1/2 by U0126 also led to a significant decrease in the level of PPARgamma, indicating that there was probably a positive cross-talk between PPARgamma and ERK 1/2 or an auto-regulatory feedback mechanism to amplify the effect of ERK1/2 on cell growth arrest and apoptosis. In addition to ERK1/2, TGZ also activated Akt. Interestingly, inhibition of ERK1/2 prevented the activation of Akt whereas suppression of Akt had no effect on ERK1/2, suggesting that Akt was not necessary for TGZ-PPARgamma-ERK pathway. However, the inhibition of Akt promoted the release of cytochrome c. Thus, the activation of Akt may have a negative effect on apoptosis induced by TGZ. Wortmannin, a PI3K inhibitor, inhibited TGZ-induced ERK1/2 and Akt activation, indicating that PI3K may function at the up-stream of ERK and Akt. In conclusion, our study has demonstrated that TGZ induced apoptosis in NCI-H23 lung cancer cells via a mitochondrial pathway and this pathway was PPARgamma-and ERK1/2-dependent. / We first investigated the effect of PPARgamma ligand TGZ on two human lung cancer cells (NCI-H23 and CRL-2066) and one human lung normal cell (CCL-202). The results showed that in consistence with the loss of cell viability, TGZ induced apoptosis in CRL-2066 and NCI-H23 cells but not in CCL-202 cells. TGZ up-regulated PPARgamma expression in all these three lung cell lines, especially in the cancer cells. In association of the time-dependent inhibition of the cell proliferation, TGZ down-regulated the expression of Bcl-w and Bcl-2 but activated ERK1/2 and p38, suggesting that the growth-inhibitory effect of TGZ is associated with the reduction of Bcl-w and Bcl-2 and the increase of ERK1/2 and p38 activation. SAPK/JNK activation assay showed a decreased activity in all these three cell lines treated by TGZ. It was also demonstrated that TGZ was able to activate PPARgamma transcriptionally. We conclude that TGZ inhibits the growth of human lung cancer cells via the induction of apoptosis, at least in part, in a PPARgamma-relevant manner. / Li Mingyue. / "June 2006." / Advisers: George Gong Chen; Anthony Ping Chuen Yim. / Source: Dissertation Abstracts International, Volume: 67-11, Section: B, page: 6202. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 174-207). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2016-06-28T18:01:18Z No. of bitstreams: 1 tiagohenrique_dissert.pdf: 1562127 bytes, checksum: 0d0b4da7bf071396a3c701c40e27e3ed (MD5) / Made available in DSpace on 2016-06-28T18:01:18Z (GMT). No. of bitstreams: 1 tiagohenrique_dissert.pdf: 1562127 bytes, checksum: 0d0b4da7bf071396a3c701c40e27e3ed (MD5) Previous issue date: 2010-07-05 / Introduction: Lung cancer is the most common malignancy in human. The average 5 years survival rate is one of the lowest among aggressive cancers, showing no significant improvement in recent years. When detected early, lung cancer has a good prognosis, but most patients present metastatic disease at the time of diagnostic, which significantly reduces survival rates. Despite all the recent advances in cancer treatment, prognostic of these patients have improved minimally. Objectives: The present study aimed to investigate the molecular profile of non-small cell lung cancer as well as new tumor makers relevant to diagnosis and prognosis of this disease. Methods: Total RNA from frozen surgical tissues was extracted using TRIZOL reagent and RNeasy FFPE kit was used for RNA extraction from formalin fixed, paraffin embedded tissue. Aiming to identify differentially expressed genes involved in lung cancer, we analyzed combined data from normal and tumor SAGE (Serial Analysis of Gene Expression) libraries available in the public domain. Proteome profiling was also analyzed in adenocarcinoma, squamous cell carcinoma and normal surgical margin samples using two-dimensional electrophoresis and mass spectrometry. Results: The statistical analysis of SAGE data indentified a subset of differentially expressed tags between normal surgical margins and adenocarcinoma libraries. Three genes displaying differential regulation in SAGE or proteomic analysis, two up- (COL3A1, CTSB) and one down-regulated (ITGB1) in neoplastic cells, were selected for real-time polymerase chain reaction (PCR) experiments using the same set of samples. Similar to the statistical results, quantitative PCR confirmed the upregulation of COL3A1 and CTSB in carcinomas when compared to tumor free tissues. Conclusion: RNA from frozen and arquived samples is appropriate for amplification experiments by real time PCR, although with lower efficiency among the last ones. Therefore, improved methods of RNA extraction in arquived tissues are suitable for Real Time quantitative RT-PCR, and may be used for gene expression profiling of paraffin embedded tissues from cancer patients. To the best of our knowledge, this is the first study reporting SAGE data analysis in lung cancer. The statistical approach as well as the proteomic evaluation were effective in identifying differentially expressed genes and proteins reportedly involved in cancer development and may be useful to disclose new tumor makers relevant to lung tumorigenesis. / Introdução: O câncer de pulmão é a neoplasia humana mais comum. As taxas de sobrevida em 5 anos estão entre as mais baixas para tumores agressivos e seus valores não têm mostrado diferenças importantes nos últimos anos. Quando detectado nos estágios precoces, o câncer de pulmão mostra bom prognóstico, mas a maioria dos pacientes apresenta doença metastática no momento do diagnóstico, o que reduz a sobrevida significativamente. Apesar de todo o progresso obtido nos últimos anos em tratamento do câncer, o prognóstico desses pacientes permanece desfavorável. Objetivos: O presente estudo teve como objetivo investigar o perfil molecular de câncer de pulmão de células não pequenas, bem como de novos marcadores tumorais relevantes para diagnóstico e prognóstico dessa doença. Métodos: O RNA total de espécimes cirúrgicos congelados foi extraído pelo método do trizol e o kit RNeasy FFPE foi utilizado para extração de RNA de tecidos fixados em formalina e emblocados em parafina. Com o objetivo de identificar genes diferencialmente expressos envolvidos em câncer de pulmão, dados combinados de bibliotecas SAGE (Serial Analysis of Gene Expression) públicas foram analisados. O perfil protéico foi também avaliado em amostras de adenocarcinoma, carcinoma epidermóide e de margens cirúrgicas normais, utilizando eletroforese bidimensional e espectrometria de massas. Resultados: A análise estatística dos dados de SAGE identificou um conjunto de tags diferencialmente expressas entre as bibliotecas de adenocarcinoma e de margens cirúrgicas. Três genes com expressão alterada na análise de SAGE e de proteômica, dois com níveis elevados (COL3A1, CTSB) e um com nível reduzido (ITGB1) em células neoplásicas, foram selecionados para experimentos de PCR (reação em cadeia da polimerase) em tempo real no mesmo conjunto de amostras. Consistente com os resultados estatísticos, a PCR quantitativa confirmou a expressão elevada de COL3A1 e CTSB em carcinomas quando comparados com o tecido livre de tumor. Conclusão: O RNA de amostras congeladas e arquivadas é adequado para amplificação por PCR em tempo real, embora exiba qualidade mais baixa nessas últimas. Portanto, métodos otimizados para tecidos arquivados permitem análises por PCR quantitativa e podem ser utilizados para avaliação do perfil transcricional de espécimes embebidos em parafina procedentes de pacientes com câncer. Este é aparentemente o primeiro estudo descrevendo a análise de dados de SAGE em câncer de pulmão. As abordagens estatística e proteômica foram efetivas em identificar genes e proteínas diferencialmente expressas envolvidas no desenvolvimento do câncer e podem revelar novos marcadores relevantes para a tumorigênese de pulmão.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2017-06-02T17:28:48Z No. of bitstreams: 1 vanessacristinalourencao_dissert.pdf: 654581 bytes, checksum: 720fe31c027c91cca25c8bf75bb27db2 (MD5) / Made available in DSpace on 2017-06-02T17:28:48Z (GMT). No. of bitstreams: 1 vanessacristinalourencao_dissert.pdf: 654581 bytes, checksum: 720fe31c027c91cca25c8bf75bb27db2 (MD5) Previous issue date: 2015-07-08 / Introduction: Stress may have a negative impact on quality of life and spirituality, independently of religion, and is considered an important coping strategy. Cancer diagnostic is a stressor that has an important impact on global or psychosocial functioning. Objective: to assess level of stress and spirituality among patients diagnosed with lung cancer. Patients and method: participants were 52 lung cancer patients receiving treatment at theInstituto do Câncer de São José do Rio Preto – SP. Data collection used a semi-structured interview, a distress thermometer, and the Functional Assessment of Chronic IllnessTherapy-Spiritual Well-Being – FACIT. Results: 57% of the patients were male and age varied between 48 and 88 years (64.29 ± 9.89); treatment duration media was 69.53 days; 82.69% reported a history of smoking and 72% had quit smoking; 88.46% have a religion and all reported to believe in God. For 61.46% of the patients, diagnosis came as “a shock”; all patients stated that religion beliefs influence health and 94% considered themselves as religious/spiritualized. 82.69% considered religiosity an important theme to be discussed by their doctor; 57.69% presented distress (negative stress) during last week, with mainly physical and emotional problems. FACIT mean score was 28.15 (sd=6.78); mean scores for sense and peace were 16,28 (sd = 3.98); mean scores for faith were 12.11 (sd=3.20). Conclusion: most patients were male and smokers, with high levels of stress. The Spirituality Scale scores were below average for sense and peace, when compared to literature and faith above average. / Introdução: O estresse pode afetar negativamente a qualidade de vida e a espiritualidade, independente da religião, é uma importante estratégia de enfrentamento. Receber um diagnóstico de câncer é um estressor que tem importante impacto sobre o funcionamento global ou biopsicossocial do indivíduo. Objetivo: Avaliar nível de estresse e espiritualidade em pacientes com diagnóstico de câncer de pulmão. Casuística e método: participaram 52 pacientes com câncer de pulmão, acompanhados na oncologia clínica do Instituto do Câncer de São José do Rio Preto – SP. Foram utilizados na coleta dos dados: Entrevista Semi-Dirigida; Termômetro de Distresse; Functional Assessment of Chronic Illness Therapy - Spiritual Well-Being - FACIT. Resultados: 57% dos pacientes eram do sexo masculino e a idade variou entre 48 e 88 anos (64,29 ± 9,89); o tempo médio de tratamento foi 69,53 dias; 82,69% relataram história de tabagismo e 72% pararam de fumar; 88,46% seguem alguma religião e todos relataram acreditar em Deus. Para 61,46% o diagnóstico veio como "um choque"; todos afirmaram que as crenças religiosas influenciam na saúde e 94% se consideram pessoas religiosas/espiritualizadas; 82,69% consideram relevante que o médico aborde a questão da religiosidade; 57,69% dos pacientes apresentaram distress (estresse negativo) na última semana, com problemas principalmente físicos e emocionais. Os dados da FACIT indicaram escore médio de 28,15 (±6,78); sentido e paz com média 16,28 (dp = 3,98) e fé com média de 12,11 (dp = 3,20). Conclusão: houve predomínio de pacientes do sexo masculino e tabagistas, com xii importante nível de estresse. A Escala de Espiritualidade apontou aspectos relacionados a "sentido e paz" abaixo da média descrita na literatura e "fé" acima da média.

De doenÃa rara no passado o cÃncer de pulmÃo, tem alcanÃado proporÃÃes alarmantes em todo o mundo, sendo considerado um problema de saÃde pÃblica mundialmente, a sua incidÃncia mudou em vÃrios paÃses nos Ãltimos anos, fato observado, principalmente, no sexo feminino. Dessa forma, a pesquisa objetivou realizar um levantamento dos estudos cientÃficos desenvolvidos acerca do tema, especialmente, da mudanÃa na distribuiÃÃo dos tipos histolÃgicos do cÃncer de pulmÃo. Estudo bibliogrÃfico, desenvolvido a partir de artigos originais, artigos de revisÃo, teses e dissertaÃÃes, revistas indexadas nas bases de dados Medline, Lilacs, NCBI, Capes, Scielo, Pub Med e Bireme, escritos na lÃngua inglesa e na portuguesa, alÃm de livros consagrados sobre a neoplasia pulmonar, totalizando 49 fontes pesquisadas. Os resultados revelaram que as taxas de novos casos e de mortalidade sÃo maiores nos paÃses desenvolvidos, especialmente nos Estados Unidos da AmÃrica (EUA) e na Europa. Embora controverso, Ã provÃvel que a mulher apresente maior susceptibilidade para o cÃncer de pulmÃo do que o homem. Essas diferenÃas entre os sexos estÃo amplamente relacionadas ao tabagismo. Contudo, tanto no homem como na mulher, a prevenÃÃo, ou seja, o combate ao tabagismo, maior fator de risco dessa neoplasia, Ã uma medida de saÃde pÃblica prioritÃria. / Lung cancer is configured with as a great public health problem in the world. From a rare disease in the past, its incidence has changed in several countries in recent years, which was observed mainly in females. Thus, the research aimed to survey the research developed on the theme, especially the change in the distribution of histological types of lung cancer. A bibliographic study, developed from original articles, review articles, theses and dissertations, journals indexed in Medline, Lilacs, NCBI, Capes, Scielo, BIREME and Pub Med data basis, written in English and Portuguese as well as books written about lung neoplasia, a total of 49 sources surveyed. The results showed that the rates of new cases and mortality are higher in developed countries, especially in the United States of America (USA) and Europe. Although controversial, it is likely that women are more susceptible to lung cancer than men. These gender differences are largely related to smoking. However, both in men and women, the prevention, in other words, the anti-smoking combat, major risk factor of this neoplasia, is a measure of public health priority.

Yung, Kam Fai. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 107-129). / Abstracts in English and Chinese. / ABSTRACT --- p.ii / 摘要 --- p.iv / ACKNOWLEDGEMENTS --- p.vi / TABLE OF CONTENTS --- p.vii / PUBLICATION --- p.ix / LIST OF TABLES --- p.x / LIST OF FIGURES --- p.xi / LIST OF ABBREVIATIONS --- p.xii / Chapter SECTION I: --- BACKGROUND --- p.1 / Chapter CHAPTER 1: --- "The biology, diagnostics and management of lung cancer" --- p.2 / Chapter 1.1 --- "Basic biology, classification and diagnostics" --- p.2 / Chapter 1.1.1 --- Epidemiology and etiology of lung cancer --- p.2 / Chapter 1.1.2 --- Clinical Presentation and Diagnostics of Lung Cancer --- p.3 / Chapter 1.2 --- Treatment of lung cancer --- p.9 / Chapter 1.2.2 --- Radiotherapy --- p.10 / Chapter 1.2.3 --- Chemotherapy --- p.11 / Chapter CHAPTER 2: --- Epidermal Growth Factor Receptor Mutations in Lung Cancer --- p.13 / Chapter 2.1 --- The Epidermal Growth Factor Receptor --- p.13 / Chapter 2.2 --- Overexpression of EGFR in NSCLC --- p.14 / Chapter 2.3 --- The development of EGFR inhibitors --- p.15 / Chapter 2.3.1 --- Monoclonal Antibodies --- p.16 / Chapter 2.3.2 --- Small-molecule inhibitors --- p.17 / Chapter 2.3.2.1 --- Gefitinib --- p.17 / Chapter 2.3.2.2 --- Erlotinib --- p.19 / Chapter 2.3.2.3 --- Other small-molecule inhibitors --- p.20 / Chapter 2.4 --- Mutations of EGFR in NSCLC --- p.21 / Chapter 2.4.1 --- Activating Mutations conferring sensitivity to tyrosine kinase inhibitors --- p.21 / Chapter 2.4.2 --- Secondary mutations associated with resistance to tyrosine kinase inhibitors --- p.23 / Chapter 2.5 --- EGFR gene amplification --- p.24 / Chapter 2.6 --- Detection of EGFR mutations --- p.25 / Chapter 2.7 --- Aim of the thesis --- p.31 / Chapter SECTION II: --- DETECTION OF EGFR MUTATIONS IN TUMOR AND PLASMA SAMPLES BY MASS SPECTROMETRY AND DIGITAL PCR --- p.33 / Chapter CHAPTER 3: --- Detection of EGFR mutations by mass spectrometric methods --- p.34 / Chapter 3.1 --- Introduction --- p.34 / Chapter 3.1.1 --- Principles of Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) --- p.34 / Chapter 3.1.2 --- The MassARRAY Homogenous MassEXTEND (hME) assay --- p.35 / Chapter 3.1.3 --- The Single-Allele Base Extension Reaction (SABER) and the Allele-Specific Base Extension Reaction (ASBER) --- p.36 / Chapter 3.2 --- Materials and Methods --- p.36 / Chapter 3.2.1 --- The protocol for the detection of EGFR exon 21 point mutation by Mass Spectrometric Methods --- p.37 / Chapter 3.3 --- Results --- p.42 / Chapter 3.4 --- Discussion --- p.49 / Chapter CHAPTER 4: --- Evaluation of the detection limit and sensitivity of the digital PCR assays --- p.51 / Chapter 4.1 --- Introduction --- p.51 / Chapter 4.1.1 --- The theoretical basis of digital PCR quantification and the relationship with the Poisson distribution --- p.51 / Chapter 4.1.2 --- Assessment of Assay Detection Limit --- p.54 / Chapter 4.1.3 --- Comparing Digital PCR with sequencing after conformation sensitive gel electrophoresis (CSGE) --- p.59 / Chapter 4.2 --- Materials and Methods --- p.59 / Chapter 4.2.1 --- Design of digital PCR assay for the detection of EGFR exon21 L858R point mutation --- p.59 / Chapter 4.2.2 --- Design of digital PCR assay for the detection of EGFR exon19 deletion --- p.60 / Chapter 4.2.3 --- The protocols of digital PCR assays for EGFR mutation detection --- p.64 / Chapter 4.2.4 --- Single molecule detection test --- p.65 / Chapter 4.2.5 --- Artificial mixtures of mutant and wild-type DNA --- p.66 / Chapter 4.2.6 --- Sequencing after CSGE --- p.66 / Chapter 4.3 --- Results --- p.67 / Chapter 4.3.1 --- Results of the single molecule detection test and artificial mixture analysis --- p.67 / Chapter 4.3.2 --- Results of CSGE and sequencing compared with digital PCR --- p.73 / Chapter 4.4 --- Discussion --- p.75 / Chapter CHAPTER 5: --- Detection of EGFR mutations in prospectively collected tumor samples of NSCLC patients --- p.77 / Chapter 5.1 --- Introduction --- p.77 / Chapter 5.2 --- Materials and Methods --- p.78 / Chapter 5.2.1 --- Sample preparation and DNA extraction of tumor tissues --- p.78 / Chapter 5.3 --- Results --- p.79 / Chapter 5.4 --- Discussion --- p.82 / Chapter CHAPTER 6: --- Detection of EGFR mutations in prospectively collected plasma samples of NSCLC patients --- p.85 / Chapter 6.1 --- Introduction --- p.85 / Chapter 6.2 --- Materials and Methods --- p.87 / Chapter 6.2.1 --- Sample preparation and DNA extraction of plasma samples --- p.87 / Chapter 6.3 --- Results --- p.88 / Chapter 6.3.1 --- Digital PCR analysis of EGFR mutations in plasma samples of NSCLC patient --- p.88 / Chapter 6.3.2 --- Variations in plasma EGFR mutation concentration after TKI treatment --- p.93 / Chapter 6.4 --- Discussion --- p.96 / Chapter SECTION III: --- CONCLUDING REMARKS --- p.100 / Chapter CHAPTER 7: --- Conclusion and future perspectives --- p.101 / Chapter 7.1 --- Mass spectrometric analysis --- p.101 / Chapter 7.2 --- Microfluidics Digital PCR --- p.102 / Chapter 7.3 --- Future perspectives --- p.105 / References --- p.107

No description available.

Thesis (Ph. D.)--University of Nevada, Reno, 1999. / Includes bibliographical references. Online version available on the World Wide Web.