Squamous cell carcinoma: clinical intraoral and demographics characteristics in a Southern Brazil population and potential interactions with lymphatic endothelial cells

Esta disserta????o foi dividida em dois trabalhos distintos, os quais podem ser resumidos da seguinte maneira: Artigo 1: O carcinoma espinocelular oral (CEO) ?? o tumor maligno mais prevalente na cavidade oral, sendo um importante problema de sa??de p??blica. O objetivo deste estudo foi avaliar o perfil cl??nico e epidemiol??gico dos casos registrados de CEO em um centro de diagn??stico cl??nico e histopatol??gico, localizado no Sul do Brasil. Oitocentos e seis indiv??duos com CEO e suas variantes histol??gicas foram inclu??dos neste estudo, num per??odo entre 1959 e 2012. As vari??veis anotadas dos arquivos foram: idade, sexo, cor da pele, s??tio, tamanho, tempo de evolu????o (relatado pelo paciente), assim como a presen??a de dor, linfonodos palp??veis, h??bitos de tabagismo e etilismo, e a profiss??o. CEO foi mais prevalente em homens (76,6%), com a maioria dos casos distribu??dos entre os 51 e 70 anos de idade (53,9%). Os s??tios mais prevalentes foram vermelh??o do l??bio inferior [23,3% (20,4; 26,4)], seguido por borda lateral/ventre de l??ngua [20,2% (17,5; 23,2)], gengiva/rebordo alveolar [18,1% (15,5; 21,0)], e assoalho bucal [14,9% (12,5; 17,5)]. Foi encontrada uma forte associa????o entre ocupa????es ao ar livre e CEO de vermelh??o de l??bio inferior. As les??es localizadas na l??ngua, gengiva/rebordo alveolar e assoalho bucal foram comumente mais dolorosas, maiores que 2 cm, a frequentemente apresentavam envolvimento de linfonodos. A maioria dos nossos resultados confirmam os dados da literatura. O autoexame bucal deveria ser recomendado e campanhas de preven????o e detec????o precoce do CEO deveriam ser realizadas periodicamente na tentativa de aumentar o sentimento pessoal em rela????o ao CEO. Artigo 2: Dentro do microambiente tumoral (MT), as c??lulas neopl??sicas est??o numa constante crosstalk com c??lulas endoteliais linf??ticas (CELs) e sangu??neas a fim de permitir o crescimento tumoral e met??stase. Supondo que haja um crosstalk entre as CELs e as c??lulas do carcinoma espinocelular (CE) que exerce um importante papel na met??stase, nosso objetivo foi identificar potenciais intera????es entre as CELs e linhagens celulares de CE, atrav??s de alguns ensaios in vitro. C??lulas endoteliais linf??ticas prim??rias adultas humanas da microvasculatura d??rmica (HMVECs) e as linhagens de CE A431, UM-SCC-1, UM-SCC-22A e UM-SCC-22B foram cultivadas nos seus meios espec??ficos. UM-SCC foram tratadas com rhIL-6, sendo a express??o de VEGF-C verificada por Elisa. Produ????o natural de IL-6 pelas HMVECs foi avaliada da mesma maneira. A presen??a de receptor de IL-6 (IL-6R) foi analisada por Western Blot nas linhagens UM-SCC. Meios condicionados(MC) das HMVECs foram preparados com diferentes tratamentos e incubados com a linhagem A431, a fim de verificar a atividade genatinol??tica das MMPs por zimografia. Nossos resultados demonstraram que h?? intera????es entre as c??lulas tumorais e as CELs, uma vez que MC-CELS foram capazes de aumentar a atividade genatinol??tica da MMP-2. Al??m disso, n??s mostramos que as CELs secretam IL-6, e diferentes linhagens de CE possuem receptores para esta citocina. Sendo assim, nossos resultados indicam potenciais intera????es entre as LECs e as c??lulas tumorais, sendo necess??rio outros estudos para elucidar as vias de sinaliza????o envolvidasThis dissertation was divided into two distinct works, which can be summarized as follows: Article 1: The oral squamous cell carcinoma (OSCC) is the most prevalent malignance in mouth, being an important public health problem. The aim of this study was to evaluate the clinical and epidemiological profile of the OSCC cases registered in a center of clinical and histopathological diagnosis, located in Southern Brazil. Eight hundred and six individuals with OSCC and its variants were included in this study, over 1959-2012 period. The variables recorded from the files were: age, gender, skin color, tumor location, size and evolution time of the lesions (referred by the patients), as well as, the presence of pain lymph nodes, habits of tobacco and alcohol, and also the profession. OSSC was more frequent in males (76.6%), with the majority of cases distributed between 51 and 70 years old (53.9%). The most prevalent sites were lower lip vermilion [23.3% (20.4; 26.4)], followed by lateral border/ventral surface of the tongue [20.2% (17.5; 23.2)], gingiva/alveolar ridge [18.1% (15.5; 21.0)], and floor of the mouth [14.9% (12.5; 17.5)]. A strong association between outdoor occupation and OSCC in lower lip vermilion was found. The OSCC lesions located in tongue, gingiva/alveolar ridge and floor of the mouth were commonly more painful, bigger than 2 cm, and frequently presenting lymph nodes involvement. Most of the results confirm the data from literature. Mouth self-examination should be recommended and campaigns of prevention and early detection of OSCC should be periodically performed in order to increase people s feelings of personal risk. Article 2: Inside the tumor microenvironment (TM) the neoplastic cells are in dynamic crosstalk with the vascular and lymphatic endothelial cells in order to allow the tumor to growth and metastasize. Hypothesizing that there is a crosstalk between lymphatic endothelial cells and tumor cells from squamous cell carcinoma (SCC) that plays an important role in metastasis, we aimed to identify potential interactions between lymphatic endothelial cells and tumor cells lines from SCCs, through some in vitro assays. Primary adult human dermal lymphatic microvascular endothelial cells (HMVECs) and the human head and neck SCC cell lines like A431, UM-SCC-1, UM-SCC-22A and UM-SCC-22B were cultured in their specific media. UM-SCC cells lines were treated with rhIL-6, being VEGF-C expression checked by Elisa. Baseline IL-6 was evaluated in HMEVCs using the same assay. Also the IL-6 receptor (IL-6R) was analyzed by Western blot in UM-SCC cells. Conditioned media from HMVECs were prepared with different treatments and incubated with SCC A431 cells, in order to verify the MMPs enzymatic activities by gelatin zymography. Our results demonstrated that there are interactions between tumor cells and LECs, since the LECs-CM were able to enhance MMP-2 gelatinolytic activity. Moreover, we showed that LECs secrete IL-6, and different SCC lines have receptors for this cytokine. Therefore, our results indicate some potential interactions between LECs and TCs, being necessary other studies to elucidate the involved signaling pathway