Introdução: O prognóstico dos carcinomas epidermóides de laringe é limitado e a taxa de sobrevida em cinco anos é menor que 70%. A relação de características clínicas e epidemiológicas tem sido investigada na sobrevida de pacientes com tumores de laringe, mas pouco se conhece sobre o efeito dos polimorfismos genéticos no prognóstico da doença. Objetivo: Estudar o papel dos polimorfismos genéticos de genes relacionados aos processos de invasão e metástase (MMP1 e MMP3), de inflamação (Interleucina 2, Interleucina 6, LTA) e reparo de DNA(XRCC1) no prognóstico do carcinoma epidermóide de laringe. Material e métodos: Coorte com 170 pacientes com carcinoma epidermóide de laringe,confirmados por exame anátomo-patológico. Os casos tiveram origem em estudo caso-controle conduzido em cinco hospitais de São Paulo, um hospital em Porto Alegre e outro em Goiânia. As informações sobre o status vital dos pacientes foram levantadas dos prontuários médicos e dos bancos de óbitos municipais e estaduais. A extração do DNA das amostras de sangue dos pacientes foi realizada pelo Instituto de Medicina Tropical da USP e a genotipagem dos polimorfismos genéticos pela Fundação Hemocentro de Ribeirão Preto da Faculdade de Medicina da USP. Resultados: Os polimorfismos genéticos estudados (MMP1 1607, MMP1 -519,MMP3 -1171, IL2 -384, IL2 114, IL6 -174, LTA 252 e XRCC1) não apresentaram efeitos com significância estatística na sobrevida global ou específica pela doença quando analisados isoladamente. Para a sobrevida global, o consumo excessivo de álcool, em g/L/dia, reduziu a sobrevida dos pacientes (80-119 g/L/dia: hazard ratio(HR)=4,0 intervalos com 95% de confiança (IC95%)=1,10-14,53; _120 g/L/dia: HR=5,6 IC95%=1,71-18,24). No modelo de Cox múltiplo, quando ajustados pelo polimorfismo genético MMP3 -1171, a sobrevida piorou para esses pacientes (80-119 g/L/dia: HR=4,9 IC95%=1,07-22,91; _120 g/L/dia: HR=6,3 IC95%=1,49-26,84). Para a sobrevida específica pela doença, o estadiamento clínico IV reduziu a sobrevida dos pacientes (HR=3,5 IC95%=1,67-7,28). No modelo de Cox múltiplo,com ajuste pelos polimorfismos genéticos IL6 -174 e MMP1 1607, a sobrevidaespecífica pela doença piorou para esses pacientes (HR=4,7 IC95%=1,38-16,25).Conclusões: Na coorte examinada, somente três dos oito polimorfismos genéticos estudados relacionaram-se com a sobrevida global e específica pela doença, porém apenas alterando o efeito dos valores dos HR brutos dos fatores consumo de álcool e estadiamento clínico, respectivamente na sobrevida global e sobrevida específica pela doença. Isoladamente, nenhum polimorfismo genético estudado interferiu na sobrevida dos pacientes com câncer de laringe. / Introduction: The prognosis of laryngeal squamous cell carcinoma is limited and survival rate is lower than 70%. The relationships between clinical and epidemiological characteristics have been fully investigated on the survival of patients with laryngeal tumors, but the effect of genetics polymorphisms on squamous cell carcinoma of larynx is not well-known. Objective: To study the role of genetic polymorphisms of genes related to the processes of invasion and metastasis (MMP1 and MMP3), inflammation (Interleukin 2, Interleukin 6, and LTA) and repair of DNA (XRCC1) in the prognosis of laryngeal squamous cell carcinoma. Material and methods: Cohort with 170 laryngeal squamous cell carcinoma patients with histological confirmation. The cases have their origin in a case-control study carried out in hospitals of Sao Paulo, Porto Alegre and Goiania. The information about vital status of patients had been raised from medical records. The extraction of DNA was carried out by Institute of Tropical Medicine of USP and genotyping was carried out by the Center of Cellular Therapy of the Hemocentro of Ribeirao Preto of Medical School of USP. Results: The studied genetic polymorphisms (MMP1 1607, MMP1 -519, MMP3 -1171, IL2 -384, IL2 114, IL6 -174, LTA 252 and XRCC1), separately analyzed, did not have any statistical significant effect on the overall and cause-specific survival. High levels of alcohol consumption (g/L/day) reduced the overall survival (80-119 g/L/day: hazard ratio(HR)=4.0 intervals with 95% of confidence (95%CI)=1.10-14.53; _120 g/L/day:HR=5.6 95%CI=1.71-18.24). Multiple Cox model revealed, when adjusted for MMP3 -1171 genetic polymorphism, lower survival for those patients (80-119g/L/day: HR=4.9 95%CI=1.07-22.91; _120 g/L/day: HR=6.3 95%CI=1.49-26.84). The clinical staging (CS) IV was a factor for low cause-specific survival (CS IV:HR=3.5 95%CI 1.67-7.28). In the multiple Cox model, adjusted for genetic polymorphism IL6 -174 and MMP1 1607, the survival of those patients droppe(HR=4.7 95%CI=1.38-16.25). Conclusions: In this cohort, only three of eight genetic polymorphisms studied were showed to be related with overall and causespecific survival, however only modifying the effect of unadjusted HR of alcohol consumption and tumor clinical staging in the overall and cause-specific survival respectively. None of the studied genetic polymorphisms, when analyzed separately,affected the survival of laryngeal cancer patients.

Thesis (PhD)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Post-operative laryngectomy patients face various physical, psychological and social challenges. The comprehensive effects of a total laryngectomy can have an adverse impact on the patient and his or her family. Because improved medical treatment can increase the life expectancy of cancer sufferers, psychosocial guidance is required over an extended period. Unfortunately, limited information on the coping strategies of such patients is available. The social work profession could benefit from greater insight into the strengths and coping mechanisms of post-operative cancer patients in order to guide them through their survivorship journey with insight and compassion. The researcher therefore aimed to explore and describe the experience of a patient and his/her close family while coping with the long-term challenges of a laryngectomy. The objectives were: (1) to discuss the nature of cancer survivorship, (2) to describe the medical aspects of and physical re-adjustments to a laryngectomy, (3) to discuss principles and strategies for coping and surviving a laryngectomy, (4) to describe the comprehensive psychosocial effects during the permanent survival phase and re-entry into society; (5) to explore the survivorship journey of laryngectomy patients and their families, and (6) to analyse and interpret data obtained from the study. Each of the survivorship phases as contextualised by Miller et al. (2008:369-374)* is discussed in the literature review. Both the ecological and the strengths perspective were utilised as the theoretical framework for this study. Principles of the strengths perspective focus on the inherent strengths that help patients cope with this traumatic life event, while the ecological perspective focuses on the utilisation of community resources in order to survive the laryngectomy experience. A combination of exploratory and descriptive designs was applied throughout the study to gain insight into the survival experience of post-operative laryngectomy patients and their families. The research question was: “What are patients’ and families’ experiences of surviving a laryngectomy with the assistance of internal and external resources within the family system and environment?” This question was addressed by combining the quantitative and qualitative research approaches. Forty-five post laryngectomy patients and fifteen family members, representing one-third of these patients, were included in the study through purposive sampling. The study period was from June 2012 to July 2013. The inclusion criteria required: (1) Patients from the service area of the selected hospital who received a total laryngectomy as surgical treatment for an advanced stage of cancer of the larynx or hypopharynx; (2) patients who were operated on not less than three months previously; (3) patients who had already completed their initial treatment and who were attending the follow-up clinic; and (4) patients who had successfully acquired trachea-oesophageal speech. Data obtained from the interviews were organised into themes. Four themes were identified: (1) the need for pre-operative information; (2) experience of physical adjustment; (3) coping and strengths used; and (4) experience of psychosocial effects of surgery and re-entry into society. These themes were divided into sub-themes and categories. The main outcome of the study was that both patients and families mobilise a combination of inner strengths and external resources to adapt to the inevitable physical changes resulting from a laryngectomy. It is therefore recommended that social workers dealing with survivorship cases utilise a combination of the ecological and strengths perspectives to create an environment in which patients can explore their own inner strengths, or to help them link to community resources whilst coping with their survivorship journey. Future research should focus on the long-term psychosocial survival of laryngectomy patients and their families, as it is likely that survivorship will increase in future; the implementation of survivorship programmes for health care professionals to equip them with skills to guide cancer survivors to full utilisation of their own strengths and available community resources; the role of pre- and primary school children/grandchildren in the rehabilitation of laryngectomy patients deserves further investigation. / AFRIKAANSE OPSOMMING: Laringektomie-pasiënte word ná hul operasie met verskeie liggaamlike, psigiese en sosiale uitdagings gekonfronteer. Die omvattende gevolge van ‘n totale laringektomie kan die pasiënt en sy of haar gesin nadelig affekteer. Aangesien verbeterde mediese behandeling die lewensverwagting van kankerpasiënte kan verleng, word psigososiale ondersteuning oor ‘n langer tydperk benodig. Ongelukkig bestaan daar baie min inligting oor hoe pasiënte kanker hanteer. Die maatskaplike werk beroep kan dus voordeel trek uit beter insig in die hanteringsmeganismes van post-operatiewe kankerpasiënte om hulle met insig en empatie deur hul oorlewingsreis te kan begelei. Die navorser het ten doel gehad om die ondervindings van die kankerpasiënt en sy/haar naby familie tydens hul langtermynhantering van ‘n laringektomie te ondersoek en te beskrywe. Verdere oogmerke van die studie was: (1) om die aard van kankeroorlewing te bespreek; (2) om die mediese aspekte van en liggaamlike aanpassing ná ‘n laringektomie te beskrywe; (3) om die beginsels en strategieë vir ‘n oorwinning oor ‘n laringektomie te bespreek; (4) om die omvattende psigososiale gevolge van die finale oorlewingsfase en hertoetrede tot die gemeenskap te beskrywe; (5) om die oorlewingsreis van die laringektomiepasiënt en sy/haar gesin te ondersoek; en (6) om die resultate van die studie te ontleed en te interpreteer. Elk van die oorlewingsfases soos deur Miller et al. (2008:369-374)* beskrywe, is in die literatuuroorsig bespreek. Die ekologiese en die sterkte-perspektiewe is tesame as teoretiese raamwerk vir die studie gebruik. Die beginsels van die sterkte-perspektief is op die inherente krag van pasiënte gemik, om te bepaal hoe hulle hierdie traumatiese lewensgebeurtenis hanteer, terwyl die ekologiese perspektief op hul aanwending van gemeenskapsbronne om die laringektomie te oorleef, fokus. ‘n Kombinasie van ondersoekende en beskrywende navorsings ontwerpe is deurgaans gebruik om insig in die oorlewingstryd van laringektomiepasiënte en hul gesinne te verkry. Die navorsingsvraag was: “Wat is pasiënte en hul gesinne se ervarings van oorlewing na ‘n laringektomie met die hulp van interne en eksterne hulpbronne in die gesinstruktuur en omgewing?” Kwantitatiewe en kwalitatiewe navorsingsmetodes is gekombineer om hierdie vraag te ondersoek. Vyf-en-veertig laringektomiepasiënte en vyftien gesinslede, wat verteenwoordigend van twee-derdes van die pasiënte was, is met behulp van ‘n doelbewuste steekproef by die studie betrek. Die studie is tussen Junie 2012 en Julie 2013 onderneem. Die insluitingskriteria was: Pasiënte uit die diensgebied van die spesifieke hospitaal wat 'n totale laringektomie as chirurgiese behandeling vir 'n gevorderde stadium van kanker van die larinks of hipofarinks ontvang het; (2) pasiënte wat hul operasie nie meer as drie maande vantevore ondergaan het nie, (3) pasiënte wat reeds hul aanvanklike behandeling voltooi het en wat die opvolgkliniek bywoon, (4) pasiënte wat tragea-esofageale spraak suksesvol bemeester het. Die data, wat deur middel van onderhoude ingesamel is, is in temas gegroepeer. Vier temas is geïdentifiseer: (1) die behoefte aan inligting voor die operasie; (2) ervaring van liggaamlike aanpassing; (3) die hantering van omstandighede en innerlike krag; en (4) ervaring van die psigososiale uitwerking van die operasie en hertoetrede tot die gemeenskap. Hierdie temas is verder in subtemas en kategorieë verdeel. Die belangrikste uitkoms van hierdie studie is dat beide pasiënte en gesinne ‘n kombinasie van hul innerlike krag en eksterne bronne aangewend het om ná die laringektomie by die onafwendbare liggaamlike veranderinge aan te pas. Daar word dus aanbeveel dat maatskaplike werkers wat kankeroorlewendes hanteer, van ‘n kombinasie van die ekologiese en die sterkte-perspektief gebruik maak om ‘n omgewing te skep waarin die pasiënt sy of haar eie innerlike krag kan ontgin, of om pasiënte te help om kontak met gemeenskapsbronne te maak terwyl hulle die oorlewingsreis baasraak. Toekomstige navorsing behoort te fokus op langtermyn psigososiale oorlewing van laringektomie pasiënte en familie met inagneming van die tendens dat kanker pasiënte se oorlewing toeneem; die implementering van opleidingsprogramme vir gesondheidswerkers te fokus om hulle met die nodige kennis toe te rus om kankeroorlewendes te begelei om hul volle krag en alle beskikbare gemeenskapsbronne te gebruik. Daarby verdien die rol van voorskoolse- en laerskoolkinders in die rehabilitasie van laringektomiepasiënte verdere ondersoek.

Thesis (M Social Work(Social Work))--University of Stellenbosch,2009. / A laryngectomy is mostly indicated as treatment for an advanced stage of cancer of the larynx, during which the patient’s voice box is removed. This operation can hold major psycho-social implications for the patient and family. To cope with the challenges in daily life, patients need social support. A lack of available literature and research regarding the role of the social worker in facilitating support groups for these patients and families was identified and motivated this research study. The goal of the study was to gain a better understanding of the laryngectomy patient’s need for support groups in a hospital setting when attending the follow-up clinic at the hospital. A combination of both the exploratory and descriptive research designs together with a combination of mainly a qualitative and to a certain extent the quantitative research approaches, was used. From this, knowledge, insight and an understanding of the need for support groups in a hospital setting from a social work perspective were obtained. Permission to conduct the proposed study was granted by the Committee for Human Research at the University of Stellenbosch. The literature study first focused on medical aspects of a laryngectomy and social work intervention services within a hospital setting as part of the multi-disciplinary approach. Second, psycho-social effects of a laryngectomy from an ecological perspective were discussed, referring to the physical, social and psychological effects of surgery upon the patient and family. Third, support and aftercare were discussed with specific reference to the role of the social worker in offering social support to the patient and family. For the empirical study, twenty laryngectomy patients from the service area of Tygerberg Hospital were involved from January 2008 to May 2008. Criteria for inclusion in the study referred to patients who had their operation not less than three months previously, attended the support group during follow-up visits at the hospital and who had successfully acquired tragea-oesophageal speech. Based on the literature review, a semi-structured questionnaire and face-to-face interview were used as research instruments to overcome the limitation of illiteracy. The results of the investigation mainly confirmed the findings of the literature study namely that laryngectomy patients can benefit from support groups in order to address their need for social support in dealing with daily life challenges. Patients indicated that they mainly needed information regarding post-operative adaptation as this was where problems were mostly experienced. The majority of patients suggested the use of support groups in this regard. Focus was placed on an exploration and description of patients’ needs for such support groups. Benefits of support groups were found to be totally compatible with the role of the social worker. The results therefore gave an indication of social work intervention services and referred to: provision of information, problem-solving, offer of social and emotional support, and promotion of rehabilitation opportunities, thereby improving the patient’s quality of life. Recommendations were aimed at social work intervention services relating to support and aftercare offered to laryngectomy patients and relatives. Future research to develop social work programmes for health care professionals in order to effectively support these patients and their families was proposed. From practical experience of support groups a study regarding the role of pre- and primary schoolchildren or grandchildren in the rehabilitation of laryngectomy patients was also suggested.

O câncer de laringe pode ser tratado com cirurgias denominadas laringectomias, nas quais toda ou parte da laringe é removida, denominadas a laringectomia total e a parcial respectivamente. Ambos os tipos de cirurgia tem um impacto importante sobre a voz. Os pacientes com problemas de voz freqüentemente referem limitações, frustrações ou até mesmo impedimentos, tanto de natureza física, como psicológica, sociais ou profissionais. Tendo em vista a relação entre a voz e a qualidade de vida (QV), o objetivo deste estudo foi investigar o impacto da disfonia na QV dos pacientes submetidos a laringectomia parcial vertical, uma vez que a principal seqüela dessa cirurgia é a disfonia. Participaram deste estudo 31 pacientes submetidos a laringectomia parcial vertical no departamento de cabeça e pescoço do Hospital Amaral Carvalho entre fevereiro de 1993 e abril de 2005. Todos os pacientes responderam ao protocolo de mensuração de qualidade de vida e voz (QVV), auto-avaliação da voz e tiveram suas vozes avaliadas, quanto ao grau da disfonia, por meio da escala GRBAS. Os resultados obtidos do protocolo QVV foram: escore total \'X BARRA\' = 77,02 \'+ OU -\' 25,10, domínio sócio-emocional \'X BARRA\' = 83,67 \'+ OU -\' 27,51 e domínio do funcionamento físico \'X BARRA\' = 72,58 \'+ OU -\' 24,74. O domínio do funcionamento físico foi que o apresentou escores mais baixos. Quanto à auto-avaliação da voz: a maioria (51,61%) dos pacientes deste estudo classificou suas vozes como boa e observou-se relação positiva entre a auto-avaliação da voz com os valores obtidos em todos os domínios do QVV, uma vez que quanto melhor foi a classificação da voz pelo paciente, maiores foram os escores do QVV indicando melhor QV. Quando correlacionados a média da escala GRBAS com os dados do protocolo QVV, não houve correlação estatisticamente significante. Embora na correlação do G (grau global da disfonia) da GRBAS com o QVV observou-se que, vozes com ) grau discreto de disfonia apresentaram maiores escores do QVV quando comparadas com vozes com graus moderado e severo, refletindo em melhor QV. Concluindo, este estudo demonstrou o impacto negativo da disfonia na QV de pacientes submetidos a laringectomia parcial vertical, uma vez que quanto maior o grau de disfonia, menores os escores do QVV. / The laryngeal cancer can be treated with surgeries denominated laryngectomies, which entire or part of larynx is removed, characterizing total laryngectomy and the partial respectively. Both types of surgery cause a very important impact in the voice. The patients with voice disorder frequently complain about limitations, frustrations or even though disabilities such as physical, psychological, social or professionals. To be considering the relationship between voice and the quality of life, the purpose of this study was research the impact of dysphonia in the quality of life in patients undergone a vertical partial laryngectomy, once the mean sequel of this surgery is the dysphonia. Thirty-one patients undergone a vertical partial laryngectomy in the Departamento de Cabeça e Pescoço do Hospital Amaral Carvalho between February 1993 and April 2005, completed the voice-quality of life QVV (translated and adapted from V-RQOL) questionnaire; self-rating voice and their voices were evaluated, as for the grade of dysphonia through GRBAS. The results obtained from: QVV total \'X BARRA\' = 77,02 e DP = 25,10, social-emotional domain \'X BARRA\' = 83,67 e DP = 27,51 and physical functional domain \'X BARRA\' = 72,58 e DP = 24,74. The physical functional domain showed the lower scores. About the voice self-rating: the majority (51,61%) of patients in this study classified their voices as good showing the positive relationship between self-rating voice with scores of QVV indicating better quality of life. When correlated the mean values of GRBAS with data QVV, statistically significant relationships were not found. In the relationship of G (grade of hoarseness) of GRBAS with the QVV, there was relationship statistically significant relation between voice and quality of life, therefore, voices with discreet grade of dysphonia showed better quality of life when compared with voices moderate and severe grades. In summary, this study demonstrated the negative impact of dysphonia in the quality of life in patients undergone a vertical partial laryngectomy, how at higher the grade of dysphonia, lower is the scores of QVV.

Os produtos naturais constituem uma das fontes mais promissoras para o descobrimento de novos ativos na terapêutica do câncer. O carcinoma de laringe é um dos mais comuns tipos de câncer envolvendo as áreas de cabeça e pescoço, e apresenta elevada taxa de morbidade e mortalidade em pacientes com estágio avançado. Terapias alternativas e/ou adjuvantes para o tratamento deste tipo de câncer representam uma necessidade emergente. Uma das alternativas mais promissoras é o desenvolvimento de nanocarreadores contendo ativos antitumorais. Araucaria angustifolia (Bert. O Kuntze) pertence à família Araucariaceae e é uma planta reconhecidamente medicinal. Seus estróbilos femininos dão origem a pinha, constituída por pinhões (sementes verdadeiras) e brácteas (sementes não desenvolvidas). O objetivo do presente estudo foi avaliar o efeito antiproliferativo do extrato aquoso de brácteas de A. angustifolia (EAA) em células tumorais de laringe HEp-2 e seus mecanismos de ação. Além disso, a possibilidade de associar o EAA a nanoesferas (NE) a fim de potencializar seu efeito antitumoral também foi investigado. A análise química por meio de Espectrometria de Massas de Alta Resolução evidenciou a presença majoritária de polifenóis no EAA. Os resultados mostraram que o EAA induziu citotoxicidade nas células tumorais HEp-2 através de dois diferentes ensaios de viabilidade celular (ensaio de MTT e de exclusão do corante Trypan blue). No entanto, o mesmo não foi capaz de induzir citotoxicidade significativa em células normais HEK-293, utilizadas como controle, indicando um efeito diferencial seletivo do EAA sobre as células tumorais. As células HEp-2 tratadas com EAA apresentaram níveis aumentados de peroxidação lipídica, danos oxidativos a proteínas e aumento da produção de ON, juntamente com depleção das defesas antioxidantes superóxido dismutase (Sod) e catalase (Cat). Além disso, o EAA induziu danos ao DNA, juntamente com fragmentação nuclear e condensação da cromatina nestas células. Alterações nos marcadores epigenéticos, como hipometilação do DNA e redução da atividade de DNMT1 foram também observadas. A exposição das células tumorais ao extrato aumentou a expressão de proteínas apoptóticas de via intrínseca mitocondrial, mediada pela ativação da proteína Bax, liberação de AIF e foi independente da ativação de p53. O EAA modificou o metabolismo energético das células tumorais, elevando os níveis de piruvato desidrogenase (PDH) e estimulando a fosforilação oxidativa mitocondrial. Embora tenha ativado a mitocôndria destas células, o mesmo causou falhas no potencial de membrana mitocondrial (ΔΨm), juntamente com diminuição dos níveis de proteínas dos complexos I e III da CTE, inibição da atividade do complexo I, produção de ERO e depleção de ATP. A associação do EAA à NE permitiu a obtenção de sistemas com tamanho de partícula inferior a 200 nm, índice de polidispersibilidade abaixo de 1, potencial zeta negativo e pH estável pelo período de 30 dias, na condição testada. No entanto, a taxa de associação obtida foi baixa (19%), indicando a necessidade de futuros estudos a fim de aumentar a eficiência de incorporação do extrato. As NE per se demonstaram capacidade de reduzir a viabilidade de células tumorais e induzir alterações redox, sensibilizando estas células, demonstrando ser um possível carreador para a vetorização de ativos antitumorais. O conjunto de dados deste estudo demonstra a potencialidade dos compostos presentes nas brácteas de A. angustifolia para o desenvolvimento de novas estratégicas terapêuticas para o câncer. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2016-12-19T15:24:14Z No. of bitstreams: 1 Tese Cátia dos Santos Branco.pdf: 11116932 bytes, checksum: 02a668c3fc8a7376e3031fe7b838d152 (MD5) / Made available in DSpace on 2016-12-19T15:24:14Z (GMT). No. of bitstreams: 1 Tese Cátia dos Santos Branco.pdf: 11116932 bytes, checksum: 02a668c3fc8a7376e3031fe7b838d152 (MD5) Previous issue date: 2016-12-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES. / Natural products are among one of the most promising fields in finding new active substances in cancer therapy. Laryngeal carcinoma is one of the most common cancers affecting the head and neck regions, and is associated with high morbidity and mortality in patients with the advanced stage. Alternative and/or complementary therapies for treating this cancer represents an emerging need. One of the most promising alternatives is the development of nanocarriers containing antitumor substances. Araucaria angustifolia (Bert. O Kuntze) belongs to Araucariaceae family and it is recognized as medicinal plant. Female strobilus originates the pinecone, which contains seeds and undeveloped seeds, commonly known as bracts. The aim of this study was to evaluate the antiproliferative effects of A. angustifolia bracts aqueous extract (AAE) in HEp-2 cancer cells and its action mechanisms. Moreover, the possibility to associate AAE in nanospheres (NS) to improve its antitumor effect was also investigated. Chemical analysis using High Resolution Mass Spectrometry (HRMS) revealed the major presence of polyphenols in AAE. The results showed that AAE induced cytotoxicity in HEp-2 cells, by using two differents approaches (MTT and Trypan blue assays). However, the extract was not able to induce significant cytotoxicity in HEK-293 normal cells used as control, indicating a selective differential effect of AEE in tumor cells. HEp-2 treated cells presented high levels of lipid peroxidation, oxidative damage to proteins and increment on NO production, along with depletion on antioxidante defenses superoxide dismutase (Sod) and catalase (Cat). In addition, AAE induced DNA damage, nuclear fragmentation and chromatin condensation in these cells. Epigenetic alterations, such as DNA hypomethylation and DNMT1 activity were also observed. Cell exposition to the AAE increased expression of proteins of the mitochondrial intrinsic pathway, via Baxtriggered, along with AIF release, and it is independent of p53 incitement. AAE changed energetic metabolism of cancer cells, increasing levels of pyruvate dehydrogenase (PDH) and stimulating mitochondrial oxidative phosphorylation. Although it enabled the mitochondria of these cells, the extract caused loss of mitochondrial membrane potential (ΔΨm), reduction on protein expression levels of complex I and III, inhibition of complex I activity, ROS generation and ATP depletion. The association of EAA to NS allowed obtaining systems with particle size lower than 200 nm, polydispersity index less than 1, negative zeta potential and pH stable for the period of 30 days, under tested condition. However, the association rate obtained was low (19%); therefore, further studies are needed to improve AAE encapsulation. The NS per se demonstrated ability to reduce the viability of tumor cells and induce redox stress, sensitizing these cells, proving to be a possible carrier for delivery of antitumor substances. The data set of this study demonstrates the potentiality of the compounds present in the A. angustifolia bracts for the development of new therapeutic strategies for cancer.

O câncer de laringe pode ser tratado com cirurgias denominadas laringectomias, nas quais toda ou parte da laringe é removida, denominadas a laringectomia total e a parcial respectivamente. Ambos os tipos de cirurgia tem um impacto importante sobre a voz. Os pacientes com problemas de voz freqüentemente referem limitações, frustrações ou até mesmo impedimentos, tanto de natureza física, como psicológica, sociais ou profissionais. Tendo em vista a relação entre a voz e a qualidade de vida (QV), o objetivo deste estudo foi investigar o impacto da disfonia na QV dos pacientes submetidos a laringectomia parcial vertical, uma vez que a principal seqüela dessa cirurgia é a disfonia. Participaram deste estudo 31 pacientes submetidos a laringectomia parcial vertical no departamento de cabeça e pescoço do Hospital Amaral Carvalho entre fevereiro de 1993 e abril de 2005. Todos os pacientes responderam ao protocolo de mensuração de qualidade de vida e voz (QVV), auto-avaliação da voz e tiveram suas vozes avaliadas, quanto ao grau da disfonia, por meio da escala GRBAS. Os resultados obtidos do protocolo QVV foram: escore total \'X BARRA\' = 77,02 \'+ OU -\' 25,10, domínio sócio-emocional \'X BARRA\' = 83,67 \'+ OU -\' 27,51 e domínio do funcionamento físico \'X BARRA\' = 72,58 \'+ OU -\' 24,74. O domínio do funcionamento físico foi que o apresentou escores mais baixos. Quanto à auto-avaliação da voz: a maioria (51,61%) dos pacientes deste estudo classificou suas vozes como boa e observou-se relação positiva entre a auto-avaliação da voz com os valores obtidos em todos os domínios do QVV, uma vez que quanto melhor foi a classificação da voz pelo paciente, maiores foram os escores do QVV indicando melhor QV. Quando correlacionados a média da escala GRBAS com os dados do protocolo QVV, não houve correlação estatisticamente significante. Embora na correlação do G (grau global da disfonia) da GRBAS com o QVV observou-se que, vozes com ) grau discreto de disfonia apresentaram maiores escores do QVV quando comparadas com vozes com graus moderado e severo, refletindo em melhor QV. Concluindo, este estudo demonstrou o impacto negativo da disfonia na QV de pacientes submetidos a laringectomia parcial vertical, uma vez que quanto maior o grau de disfonia, menores os escores do QVV. / The laryngeal cancer can be treated with surgeries denominated laryngectomies, which entire or part of larynx is removed, characterizing total laryngectomy and the partial respectively. Both types of surgery cause a very important impact in the voice. The patients with voice disorder frequently complain about limitations, frustrations or even though disabilities such as physical, psychological, social or professionals. To be considering the relationship between voice and the quality of life, the purpose of this study was research the impact of dysphonia in the quality of life in patients undergone a vertical partial laryngectomy, once the mean sequel of this surgery is the dysphonia. Thirty-one patients undergone a vertical partial laryngectomy in the Departamento de Cabeça e Pescoço do Hospital Amaral Carvalho between February 1993 and April 2005, completed the voice-quality of life QVV (translated and adapted from V-RQOL) questionnaire; self-rating voice and their voices were evaluated, as for the grade of dysphonia through GRBAS. The results obtained from: QVV total \'X BARRA\' = 77,02 e DP = 25,10, social-emotional domain \'X BARRA\' = 83,67 e DP = 27,51 and physical functional domain \'X BARRA\' = 72,58 e DP = 24,74. The physical functional domain showed the lower scores. About the voice self-rating: the majority (51,61%) of patients in this study classified their voices as good showing the positive relationship between self-rating voice with scores of QVV indicating better quality of life. When correlated the mean values of GRBAS with data QVV, statistically significant relationships were not found. In the relationship of G (grade of hoarseness) of GRBAS with the QVV, there was relationship statistically significant relation between voice and quality of life, therefore, voices with discreet grade of dysphonia showed better quality of life when compared with voices moderate and severe grades. In summary, this study demonstrated the negative impact of dysphonia in the quality of life in patients undergone a vertical partial laryngectomy, how at higher the grade of dysphonia, lower is the scores of QVV.

Os produtos naturais constituem uma das fontes mais promissoras para o descobrimento de novos ativos na terapêutica do câncer. O carcinoma de laringe é um dos mais comuns tipos de câncer envolvendo as áreas de cabeça e pescoço, e apresenta elevada taxa de morbidade e mortalidade em pacientes com estágio avançado. Terapias alternativas e/ou adjuvantes para o tratamento deste tipo de câncer representam uma necessidade emergente. Uma das alternativas mais promissoras é o desenvolvimento de nanocarreadores contendo ativos antitumorais. Araucaria angustifolia (Bert. O Kuntze) pertence à família Araucariaceae e é uma planta reconhecidamente medicinal. Seus estróbilos femininos dão origem a pinha, constituída por pinhões (sementes verdadeiras) e brácteas (sementes não desenvolvidas). O objetivo do presente estudo foi avaliar o efeito antiproliferativo do extrato aquoso de brácteas de A. angustifolia (EAA) em células tumorais de laringe HEp-2 e seus mecanismos de ação. Além disso, a possibilidade de associar o EAA a nanoesferas (NE) a fim de potencializar seu efeito antitumoral também foi investigado. A análise química por meio de Espectrometria de Massas de Alta Resolução evidenciou a presença majoritária de polifenóis no EAA. Os resultados mostraram que o EAA induziu citotoxicidade nas células tumorais HEp-2 através de dois diferentes ensaios de viabilidade celular (ensaio de MTT e de exclusão do corante Trypan blue). No entanto, o mesmo não foi capaz de induzir citotoxicidade significativa em células normais HEK-293, utilizadas como controle, indicando um efeito diferencial seletivo do EAA sobre as células tumorais. As células HEp-2 tratadas com EAA apresentaram níveis aumentados de peroxidação lipídica, danos oxidativos a proteínas e aumento da produção de ON, juntamente com depleção das defesas antioxidantes superóxido dismutase (Sod) e catalase (Cat). Além disso, o EAA induziu danos ao DNA, juntamente com fragmentação nuclear e condensação da cromatina nestas células. Alterações nos marcadores epigenéticos, como hipometilação do DNA e redução da atividade de DNMT1 foram também observadas. A exposição das células tumorais ao extrato aumentou a expressão de proteínas apoptóticas de via intrínseca mitocondrial, mediada pela ativação da proteína Bax, liberação de AIF e foi independente da ativação de p53. O EAA modificou o metabolismo energético das células tumorais, elevando os níveis de piruvato desidrogenase (PDH) e estimulando a fosforilação oxidativa mitocondrial. Embora tenha ativado a mitocôndria destas células, o mesmo causou falhas no potencial de membrana mitocondrial (ΔΨm), juntamente com diminuição dos níveis de proteínas dos complexos I e III da CTE, inibição da atividade do complexo I, produção de ERO e depleção de ATP. A associação do EAA à NE permitiu a obtenção de sistemas com tamanho de partícula inferior a 200 nm, índice de polidispersibilidade abaixo de 1, potencial zeta negativo e pH estável pelo período de 30 dias, na condição testada. No entanto, a taxa de associação obtida foi baixa (19%), indicando a necessidade de futuros estudos a fim de aumentar a eficiência de incorporação do extrato. As NE per se demonstaram capacidade de reduzir a viabilidade de células tumorais e induzir alterações redox, sensibilizando estas células, demonstrando ser um possível carreador para a vetorização de ativos antitumorais. O conjunto de dados deste estudo demonstra a potencialidade dos compostos presentes nas brácteas de A. angustifolia para o desenvolvimento de novas estratégicas terapêuticas para o câncer. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, CAPES. / Natural products are among one of the most promising fields in finding new active substances in cancer therapy. Laryngeal carcinoma is one of the most common cancers affecting the head and neck regions, and is associated with high morbidity and mortality in patients with the advanced stage. Alternative and/or complementary therapies for treating this cancer represents an emerging need. One of the most promising alternatives is the development of nanocarriers containing antitumor substances. Araucaria angustifolia (Bert. O Kuntze) belongs to Araucariaceae family and it is recognized as medicinal plant. Female strobilus originates the pinecone, which contains seeds and undeveloped seeds, commonly known as bracts. The aim of this study was to evaluate the antiproliferative effects of A. angustifolia bracts aqueous extract (AAE) in HEp-2 cancer cells and its action mechanisms. Moreover, the possibility to associate AAE in nanospheres (NS) to improve its antitumor effect was also investigated. Chemical analysis using High Resolution Mass Spectrometry (HRMS) revealed the major presence of polyphenols in AAE. The results showed that AAE induced cytotoxicity in HEp-2 cells, by using two differents approaches (MTT and Trypan blue assays). However, the extract was not able to induce significant cytotoxicity in HEK-293 normal cells used as control, indicating a selective differential effect of AEE in tumor cells. HEp-2 treated cells presented high levels of lipid peroxidation, oxidative damage to proteins and increment on NO production, along with depletion on antioxidante defenses superoxide dismutase (Sod) and catalase (Cat). In addition, AAE induced DNA damage, nuclear fragmentation and chromatin condensation in these cells. Epigenetic alterations, such as DNA hypomethylation and DNMT1 activity were also observed. Cell exposition to the AAE increased expression of proteins of the mitochondrial intrinsic pathway, via Baxtriggered, along with AIF release, and it is independent of p53 incitement. AAE changed energetic metabolism of cancer cells, increasing levels of pyruvate dehydrogenase (PDH) and stimulating mitochondrial oxidative phosphorylation. Although it enabled the mitochondria of these cells, the extract caused loss of mitochondrial membrane potential (ΔΨm), reduction on protein expression levels of complex I and III, inhibition of complex I activity, ROS generation and ATP depletion. The association of EAA to NS allowed obtaining systems with particle size lower than 200 nm, polydispersity index less than 1, negative zeta potential and pH stable for the period of 30 days, under tested condition. However, the association rate obtained was low (19%); therefore, further studies are needed to improve AAE encapsulation. The NS per se demonstrated ability to reduce the viability of tumor cells and induce redox stress, sensitizing these cells, proving to be a possible carrier for delivery of antitumor substances. The data set of this study demonstrates the potentiality of the compounds present in the A. angustifolia bracts for the development of new therapeutic strategies for cancer.

Lo, Chun Shan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 71-80). / Abstracts in English and Chinese. / Abstract --- p.i / Chinese abstract --- p.iii / Acknowledgements --- p.iv / List of figures --- p.vi / Abbreviations --- p.vii / Contents --- p.viii / Chapter Chapter One --- General Introduction --- p.Page / Chapter 1.1 --- Background --- p.1 / Chapter 1.2 --- Human papillomavirus infection at the larynx --- p.2 / Chapter 1.2.1 --- Biology of human papillomavirus --- p.4 / Chapter 1.2.2 --- HPV E6 protein --- p.5 / Chapter 1.2.3 --- HPV E7 protein --- p.7 / Chapter 1.3 --- Apoptosis --- p.9 / Chapter 1.3.1 --- Apoptosis signaling pathways --- p.11 / Chapter 1.4 --- Transcription factor: Nuclear factor -kB --- p.14 / Chapter 1.4.1 --- Overview of the NF-kB signaling pathway --- p.14 / Chapter 1.4.2 --- Regulation of NF-kB signaling --- p.16 / Chapter 1.4.3 --- Roles of NF-kB in cancers --- p.19 / Chapter 1.5 --- Pteris semipinnata L extract: ent-11 -hydroxy-15-oxo-kaur-16-en-19-oic-acid (5F) --- p.21 / Chapter 1.6 --- Objectives --- p.22 / Chapter Chapter Two --- Materials and Methods / Chapter 2.1 --- Cell culture --- p.24 / Chapter 2.2 --- Cell proliferation analysis --- p.24 / Chapter 2.3 --- Western Blotting --- p.26 / Chapter 2.3.1 --- Total protein extraction --- p.26 / Chapter 2.3.2 --- Nuclear and cytoplasmic protein extraction --- p.26 / Chapter 2.3.3 --- Quantification of protein concentration --- p.27 / Chapter 2.3.4 --- Sodium dodecyl sulfate - polyacylamide gel electrophoresis (SDS-PAGE) and protein transfer --- p.28 / Chapter 2.3.5 --- Immunoblotting --- p.29 / Chapter 2.4 --- NF-kB Luciferase Assay --- p.29 / Chapter 2.5 --- Annexin V apoptosis assay --- p.31 / Chapter 2.6 --- mRNA expression analyses --- p.33 / Chapter 2.6.1 --- RNA extraction --- p.33 / Chapter 2.6.2 --- Reverse Transcription --- p.33 / Chapter 2.6.3 --- Polymerase Chain Reaction --- p.34 / Chapter 2.7 --- Antibodies --- p.35 / Chapter Chapter Three --- Results / Chapter 3.1 --- "Anti-proliferation effect of 5F on laryngeal cancer cells UMSCC11A, UMSCC12 and HEp-2 cells" --- p.36 / Chapter 3.2 --- Suppression by 5F in HEp-2 of mRNA and protein expression levels in HPV18 E7 while the expression level of HPV18 E6 was not altered --- p.38 / Chapter 3.3 --- Quantification of 5F-induced apoptosis in laryngeal cancer cells by Annexin V assay --- p.40 / Chapter 3.4 --- Morphological changes in laryngeal cancer cells induced by 5F --- p.41 / Chapter 3.5 --- "Cleavage of poly (ADP-ribose) polymerase (PARP) and pro-caspase-3 induced by 5F in UMSCC11 A, UMSCC12 and HEp-2 cell lines" --- p.47 / Chapter 3.6 --- "Down-regulation of TNF-α-induced NF-kB subunit p65 and p50 nuclear translocations in UMSCC11 A, UMSCC12 and HEp-2 by 5F" --- p.47 / Chapter 3.7 --- Dose-dependent inhibition of 5F on NF-kB transcriptional activity measured by luciferase assay --- p.53 / Chapter 3.8 --- Partial inhibition of TNF-α induced kBα degradation by 5F in UMSCC11A but not in UMSCC12 and HEp-2 --- p.56 / Chapter 3.9 --- Cell proliferation inhibition and apoptosis induction by Bay (11-7082) in laryngeal cancer cells --- p.56 / Chapter 3.10 --- Differential basal nuclear translocation of p65 and p50 in laryngeal cancer cell lines --- p.57 / Chapter 3.11 --- 5F regulated NF-kB target gene expression --- p.58 / Chapter Chapter Four --- Discussions --- p.64 / Reference --- p.71 / Appendix / Appendix 1 Map of pLuc- NF-kB plasmid --- p.81

Os objetivos deste estudo foram identificar os fatores de risco para complicações pulmonares pós-operatórias em pacientes laringectomizados por carcinoma espinocelular de laringe, assim como os fatores de risco para apresentação de metástases pulmonares e os fatores de risco para apresentação de segundo tumores primários em pulmão destes pacientes no período de 1985 a 1996. O estudo consistiu em uma análise retrospectiva de 291 pacientes admitidos no Hospital do Câncer A. C. Camargo, no período de 1985 a 1996, portadores de carcinoma espinocelular de laringe, submetidos a tratamento cirúrgico com intenção curativa seguido ou não de radioterapia. Foram analisados as variáveis demográficas, as comorbidades, a localização do sítio primário, o estadiamento clínico, o tratamento do tumor primário e do pescoço, o tratamento radioterápico, a diferenciação celular, as margens cirúrgicas, as recidivas locais, as recidivas regionais, a presença de complicações maiores, de complicações menores, as metástases pulmonares e a presença de múltiplos tumores primários pulmonares. O teste de associação do qui-quadrado foi utilizado para análise univariada descritiva das diversas variáveis comparando-se os grupos com complicação pulmonar e sem complicação pulmonar. A análise multivariada através da regressão logística foi utilizada na determinação dos fatores de risco para apresentação de metástases pulmonares e múltiplos tumores primário pulmonares. A incidência de complicações pulmonares foi de 31,3%, foram identificados como fatores de risco para complicações pulmonares somente a epiglote (p=0,004; RR 2,1), tendo a variável gênero associação marginal (p=0,081; RR 2,8). As metástases pulmonares tiveram incidência de 7,2%, na análise univariada foram identificados como fatores de risco o estadiamento N (p=0,032), diferenciação histológica (p=0,004), margens cirúrgicas (p=0,017) e recidivas locoregionais (0,002). Os múltiplos tumores primários pulmonares apresentaram incidência de 3,1% e na análise univariada foram identificados como fatores de risco o estadiamento N (p=0,048) e sítio aritenóide (p=0,001). Na análise multivariada foram significativos somente a diferenciação histológica: moderamente diferenciado (p=0,007; RR 2,9) e pouco diferenciado (p=0,032; RR 4,0); e as margens cirúrgicas: exíguas (p=0,003; RR 6,4) para apresentação de metástases pulmonares e múltiplos tumores pulmonares. Este estudo demonstra a importância do estadiamento clínico como fator de risco para complicações pulmonares, metástases à distância e múltiplos tumores primário em pulmão. Os fatores de risco determinantes para aparecimento de metástases pulmonares e múltiplos tumores primários pulmonares foram a diferenciação histológica e as margens cirúrgicas / Objective: To identify the risk factors to postoperative pulmonary complications in laryngeal cancer patients submitted to surgical treatment, the risk factors to development of lung metastasis and second lung primary tumor. Patients and Methods: Retrospective study of a cohort of 291 patients admitted at Hospital do Câncer A.C.Camargo from January, 1985 to December 1996. All patients were submitted to some kind of laryngectomy with curative intent as part of treatment of a proven laryngeal cancer, followed or not by radiotherapy. The following variables were analized: demographic, comorbidities, primary site, clinical stage, primary and neck surgical treatment, histopathologic differentiation grade, surgical margins, recurrences, postoperative pulmonary complications, lung metastasis and second lung primary tumor. The univariate and multivariate analysis were utilized to built the model to predict the risks factors and the factors of prognostic significance. Results: The overall pulmonary complications incidence were 31,3%, epiglottis were identified as significant single risk factor to pulmonary complications (p=0.004; RR 2,1). Lung metastasis had 7,2% incidence to this, the risk factors were N stage (p=0.032 ), histopathologic differentiation grade (p=0.004), surgical margins (p=0.017) and locoregional recurrence (p=0.002). The second lung primary tumor incidence were 3,1%, univariate analysis showed N stage (p=0,048) and arithenoid site (p=0,001) as significant risk factors. The multivariate analysis showed the histopathologic differentiation: moderate grade (p=0.007 RR 2,9) and poor grade (p=0.032 RR 4,0) and surgical margins: close (p=0.003 RR 6,4) as prognostic factors to deveopment of lung metastasis and second lung primary tumor. Conclusions: This study showed the clinical stage importance as risk factor to development of postoperative pulmonary complication, lung metastasis and second lung primary tumor in laryngectomy cancer patients. The prognostic factors associated with lung metastasis and second lung primary tumor were the histopathologic differentiation and the surgical margins

Os objetivos deste estudo foram identificar os fatores de risco para complicações pulmonares pós-operatórias em pacientes laringectomizados por carcinoma espinocelular de laringe, assim como os fatores de risco para apresentação de metástases pulmonares e os fatores de risco para apresentação de segundo tumores primários em pulmão destes pacientes no período de 1985 a 1996. O estudo consistiu em uma análise retrospectiva de 291 pacientes admitidos no Hospital do Câncer A. C. Camargo, no período de 1985 a 1996, portadores de carcinoma espinocelular de laringe, submetidos a tratamento cirúrgico com intenção curativa seguido ou não de radioterapia. Foram analisados as variáveis demográficas, as comorbidades, a localização do sítio primário, o estadiamento clínico, o tratamento do tumor primário e do pescoço, o tratamento radioterápico, a diferenciação celular, as margens cirúrgicas, as recidivas locais, as recidivas regionais, a presença de complicações maiores, de complicações menores, as metástases pulmonares e a presença de múltiplos tumores primários pulmonares. O teste de associação do qui-quadrado foi utilizado para análise univariada descritiva das diversas variáveis comparando-se os grupos com complicação pulmonar e sem complicação pulmonar. A análise multivariada através da regressão logística foi utilizada na determinação dos fatores de risco para apresentação de metástases pulmonares e múltiplos tumores primário pulmonares. A incidência de complicações pulmonares foi de 31,3%, foram identificados como fatores de risco para complicações pulmonares somente a epiglote (p=0,004; RR 2,1), tendo a variável gênero associação marginal (p=0,081; RR 2,8). As metástases pulmonares tiveram incidência de 7,2%, na análise univariada foram identificados como fatores de risco o estadiamento N (p=0,032), diferenciação histológica (p=0,004), margens cirúrgicas (p=0,017) e recidivas locoregionais (0,002). Os múltiplos tumores primários pulmonares apresentaram incidência de 3,1% e na análise univariada foram identificados como fatores de risco o estadiamento N (p=0,048) e sítio aritenóide (p=0,001). Na análise multivariada foram significativos somente a diferenciação histológica: moderamente diferenciado (p=0,007; RR 2,9) e pouco diferenciado (p=0,032; RR 4,0); e as margens cirúrgicas: exíguas (p=0,003; RR 6,4) para apresentação de metástases pulmonares e múltiplos tumores pulmonares. Este estudo demonstra a importância do estadiamento clínico como fator de risco para complicações pulmonares, metástases à distância e múltiplos tumores primário em pulmão. Os fatores de risco determinantes para aparecimento de metástases pulmonares e múltiplos tumores primários pulmonares foram a diferenciação histológica e as margens cirúrgicas / Objective: To identify the risk factors to postoperative pulmonary complications in laryngeal cancer patients submitted to surgical treatment, the risk factors to development of lung metastasis and second lung primary tumor. Patients and Methods: Retrospective study of a cohort of 291 patients admitted at Hospital do Câncer A.C.Camargo from January, 1985 to December 1996. All patients were submitted to some kind of laryngectomy with curative intent as part of treatment of a proven laryngeal cancer, followed or not by radiotherapy. The following variables were analized: demographic, comorbidities, primary site, clinical stage, primary and neck surgical treatment, histopathologic differentiation grade, surgical margins, recurrences, postoperative pulmonary complications, lung metastasis and second lung primary tumor. The univariate and multivariate analysis were utilized to built the model to predict the risks factors and the factors of prognostic significance. Results: The overall pulmonary complications incidence were 31,3%, epiglottis were identified as significant single risk factor to pulmonary complications (p=0.004; RR 2,1). Lung metastasis had 7,2% incidence to this, the risk factors were N stage (p=0.032 ), histopathologic differentiation grade (p=0.004), surgical margins (p=0.017) and locoregional recurrence (p=0.002). The second lung primary tumor incidence were 3,1%, univariate analysis showed N stage (p=0,048) and arithenoid site (p=0,001) as significant risk factors. The multivariate analysis showed the histopathologic differentiation: moderate grade (p=0.007 RR 2,9) and poor grade (p=0.032 RR 4,0) and surgical margins: close (p=0.003 RR 6,4) as prognostic factors to deveopment of lung metastasis and second lung primary tumor. Conclusions: This study showed the clinical stage importance as risk factor to development of postoperative pulmonary complication, lung metastasis and second lung primary tumor in laryngectomy cancer patients. The prognostic factors associated with lung metastasis and second lung primary tumor were the histopathologic differentiation and the surgical margins