No description available.

No description available.

Yes / Cells encapsulation is a micro-technology widely applied in cell and tissue research, tissue transplantation, and regenerative medicine. In this paper, we proposed a growth of microtissue model for the human keratinocytes (HaCaT) cell line and an oral squamous cell carcinoma (OSCC) cell line (ORL-48) based on a simple aerosol microencapsulation technique. At an extrusion rate of 20 μL/min and air flow rate of 0.3 L/min programmed in the aerosol system, HaCaT and ORL-48 cells in alginate microcapsules were encapsulated in microcapsules with a diameter ranging from 200 to 300 μm. Both cell lines were successfully grown into microtissues in the microcapsules of alginate within 16 days of culture. The microtissues were characterized by using a live/dead cell viability assay, field emission-scanning electron microscopy (FE-SEM), fluorescence staining, and cell re-plating experiments. The microtissues of both cell types were viable after being extracted from the alginate membrane using alginate lyase. However, the microtissues of HaCaT and ORL-48 demonstrated differences in both nucleus size and morphology. The microtissues with re-associated cells in spheroids are potentially useful as a cell model for pharmacological studies. / Malaysia Ministry of Education (Fundamental Research Grant Scheme, FRGS Vot. 1482 and IGSP Vot. 679).

Yes / Background - Previous studies have demonstrated that a number of HOX genes, a family of transcription factors with key roles in early development, are up-regulated in head and neck squamous cell carcinoma (HNSCC) and other cancers. The loci of several Homeobox (HOX) genes also contain microRNAs (miRs), including miR-196a. Methods - Global miR expression and expression of all 39 HOX genes in normal oral keratinocytes (NOKs), oral pre-malignant (OPM) and HNSCC cells was assessed by expression microarray and qPCR and in tissues by immunohistochemistry (IHC) and qPCR of laser microdissected (LCM) tissues. Expression of miR196a and HOXB9 was reduced using anti-miR-196a and siRNA, respectively. Expression microarray profiles of anti-miR196a and pre-miR196a transfected cells were compared to parental cells in order to identify novel targets of miR- 196a. Putative miR196a targets were validated by qPCR and were confirmed as binding to the 3’UTR of miR196a by a dual luciferase reporter assay combined with mutational analysis of the miR-196a binding site. Results - miR-196a and HOXB9 are highly expressed in HNSCC compared to NOKs, a pattern also seen in HNSCC tissues by HOXB9 IHC and qPCR of miR-196a in LCM tissue. Knock-down of miR-196a expression decreased HNSCC cell migration, invasion and adhesion to fibronectin, but had no effect on proliferation. Furthermore, knock-down of HOXB9 expression decreased migration, invasion and proliferation but did not alter adhesion. We identified a novel primary mRNA transcript containing HOXB9 and miR196a-1 as predicted from in-silico analysis. Expression array analysis identified a number of miR196a targets, including MAMDC2 and HOXC8. We confirmed that MAMDC2 is a novel miR-196a target using a dual luciferase reporter assay with the effect abolished on mutation of the binding site. Conclusions - These results show that miR-196a and HOXB9 are overexpressed, perhaps co-ordinately, as HNSCC develops and exert a pro-tumourigenic phenotype in HNSCC and OPM cells.

Diss. (sammanfattning) Umeå : Univ., 2003. / Härtill 4 uppsatser.

INTRODUÇÃO: A ressecção do ramo ascendente da mandíbula foi, durante várias décadas, considerada o tratamento de eleição para os tumores da loja amigdalina e região retromolar, independente do grau de acometimento do osso mandibular, ocasionando um déficit funcional e estético considerável, muitas vezes, com prejuízos irreparáveis à qualidade de vida, às vezes desnecessário. Assim, a ressecção marginal do osso mandibular surgiu como uma alternativa de tratamento viável, uma vez que a manutenção de um segmento do ramo mandibular, em lesões sem comprometimento ósseo, não aumenta os índices de recidiva, tampouco compromete os princípios de radicalidade oncológica. OBJETIVO E MÉTODOS: Por meio de estudo retrospectivo de Outubro de 1994 a Dezembro de 2001, foram comparados 42 pacientes portadores de tumores avançados de região retromolar e loja amigdalina, sendo 20 deles submetidos a ressecção marginal do osso mandibular e 22 submetidos a ressecção do ramo ascendente da mandíbula, em relação a complicações, seqüela de procedimentos, recidiva locorregional e sobrevida. RESULTADOS: Dos 20 pacientes tratados com mandibulectomia marginal, avaliados por um período de 9 a 60 meses, sete (35%) pacientes morreram com doença, com sobrevida mínima de 09 meses, 3 por recidiva local, 3 por recidiva regional e 1 por recidiva locorregional. Um paciente morreu no pós-operatório imediato. Na avaliação da peça cirúrgica encontramos todas as margens livres, considerada exígua em profundidade em dois pacientes, um deles falecido por recidiva local. Houve disseminação linfonodal em 15 pacientes sendo com ruptura extracapsular em 4, encontrada em 2 pacientes com recidiva regional. O controle locorregional foi obtido em 63% dos pacientes. Dos 22 pacientes tratados com ressecção segmentar do osso mandibular, com intervalo de seguimento de 14 a 60 meses, 8 (36,4%) morreram pela doença, com sobrevida mínima de 9 meses, 5 por recidiva local e 3 por recidiva à distância. Um paciente morreu no pós-operatório imediato. As margens foram livres em 20 pacientes e, em 3 exíguas, um deles falecido por recidiva local. Houve disseminação linfonodal em 12 pacientes com ruptura extracapsular em 7 pacientes. O controle locorregional foi obtido em 61% dos pacientes. Na curva de análise de sobrevida, pelo método de Kaplan-Meier, o grupo tratado com mandibulectomia marginal apresentou uma taxa de 42%, com intervalo de 31 a 52 meses, erro padrão de 5 meses e intervalo de confiança de 95% e o grupo tratado com ressecção segmentar 38% com intervalo de 27 a 48 meses, erro padrão de 5 meses, um intervalo de confiança de 95%. A comparação pelo teste de Log Rank, não paramétrico apresentou p<0,8329 e pelo teste t-Student p< 0,621 ambos não significantes. As principais complicações foram a infecção local em 5 (11,9%) pacientes e a fístula orocutânea em 4 (9,5%). Houve uma fratura da placa de titânio, dois pacientes evoluíram com osteorradionecrose e nove com disfunção da articulação têmporo-mandibular. CONCLUSÕES: Não houve diferenças estatisticamente significantes entre os grupos de ressecção marginal e segmentar nos critérios analisados. Portanto, a conservação do ramo ascendente da mandíbula, em lesões que não apresentem envolvimento mandibular, mesmo avançadas, não aumenta o índice de recidiva. / INTRODUCTION: For several decades, resection of the ascending ramus of the mandible was considered to be mandatory for the treatment of tonsil and retromolar trigone tumours, independent from the damage degree of the mandibular bone, causing considerable functional and aesthetic deficit, many times with irreparable quality of life loss, sometimes unnecessary. Thus, marginal resection of the mandibular bone appeared as a feasible alternative treatment, since maintaining a segment of the mandibular ramus in lesions without bone involvement did not increase the recurrence indexes or compromise the principles of oncological radicalness. OBJECTIVES AND METHODS: Through a retrospective study from October 1994 to December 2001, 42 patients with advanced retromolar and tonsil tumors were compared, 20 undergoing marginal resection of the mandibular bone and 22 undergoing segmental resection of the ascending ramus of the mandible, with regard complications, injury originated from the procedure, locoregional recurrence and survival. RESULTS: From the 20 patients undergoing to marginal mandibulectomy, assessed for a period of 09 to 60 months, seven (35%) patients died of the disease, with a minimal survival of 9 months: 3 due to local recurrence, 3 due to regional recurrence and 1 due to local and regional recurrence. One patient died in the immediate postoperative period. When assessing the surgical part, all the margins were found to be free and in two patients, were considered to be of little depth, one of them being found in one of the patients that died from local recurrence. There was lymph nodal dissemination in 15 patients, with capsular rupture in 4, of which two presented regional recurrence. The locoregional control was obtained in 63% of the patients. From the 22 patients undergoing to segmental resection of the mandibular bone, with a follow-up varying from 14 to 60 months, 8 (36.4%) died of the disease, with a minimum survival of 9 months, 5 due to local recurrence and 3 due to distant recurrence. One patient died in the immediate postoperative period. The surgical margins were considered free from disease in 20 patients and, in three they were too small, one patient died of local recurrence. There was lymph nodal dissemination in 12 patients, and 7 presented capsular rupture. The locoregional control was obtained in 61% of the patients. In the survival analysis curve, by the Kaplan-Meier method, the group submitted to marginal mandibulectomy presented with a rate of 42%, with an interval of 31 to 52 months, standard error of 5 months and confidence interval of 95%, and the group submitted to segmentary resection, 38% with an interval of 27 to 48 months, a standard error of 5 months, and a confidence interval of 95%. The comparison using the non-parametric Log-Rank test presented p<0.8326 and the t-Student test p< 0.621, both not statistically significant. The main complications were local infection in 5 (11.9%) patients and oro-cutaneous fistula in 4 (9.5%). There was one titanium plate fracture, two patients developed osteoradionecrosis and nine temporo-mandibular dysfunction. CONCLUSIONS: There weren t statistically significant differences between the marginal and the segmental resection groups in relation to the analyzed criteria. So, the preservation of the ascending ramus of the mandible, in lesions which do not present mandible commitment, even if not advanced, doesn t increase the recurrence rate.

INTRODUÇÃO: A ressecção do ramo ascendente da mandíbula foi, durante várias décadas, considerada o tratamento de eleição para os tumores da loja amigdalina e região retromolar, independente do grau de acometimento do osso mandibular, ocasionando um déficit funcional e estético considerável, muitas vezes, com prejuízos irreparáveis à qualidade de vida, às vezes desnecessário. Assim, a ressecção marginal do osso mandibular surgiu como uma alternativa de tratamento viável, uma vez que a manutenção de um segmento do ramo mandibular, em lesões sem comprometimento ósseo, não aumenta os índices de recidiva, tampouco compromete os princípios de radicalidade oncológica. OBJETIVO E MÉTODOS: Por meio de estudo retrospectivo de Outubro de 1994 a Dezembro de 2001, foram comparados 42 pacientes portadores de tumores avançados de região retromolar e loja amigdalina, sendo 20 deles submetidos a ressecção marginal do osso mandibular e 22 submetidos a ressecção do ramo ascendente da mandíbula, em relação a complicações, seqüela de procedimentos, recidiva locorregional e sobrevida. RESULTADOS: Dos 20 pacientes tratados com mandibulectomia marginal, avaliados por um período de 9 a 60 meses, sete (35%) pacientes morreram com doença, com sobrevida mínima de 09 meses, 3 por recidiva local, 3 por recidiva regional e 1 por recidiva locorregional. Um paciente morreu no pós-operatório imediato. Na avaliação da peça cirúrgica encontramos todas as margens livres, considerada exígua em profundidade em dois pacientes, um deles falecido por recidiva local. Houve disseminação linfonodal em 15 pacientes sendo com ruptura extracapsular em 4, encontrada em 2 pacientes com recidiva regional. O controle locorregional foi obtido em 63% dos pacientes. Dos 22 pacientes tratados com ressecção segmentar do osso mandibular, com intervalo de seguimento de 14 a 60 meses, 8 (36,4%) morreram pela doença, com sobrevida mínima de 9 meses, 5 por recidiva local e 3 por recidiva à distância. Um paciente morreu no pós-operatório imediato. As margens foram livres em 20 pacientes e, em 3 exíguas, um deles falecido por recidiva local. Houve disseminação linfonodal em 12 pacientes com ruptura extracapsular em 7 pacientes. O controle locorregional foi obtido em 61% dos pacientes. Na curva de análise de sobrevida, pelo método de Kaplan-Meier, o grupo tratado com mandibulectomia marginal apresentou uma taxa de 42%, com intervalo de 31 a 52 meses, erro padrão de 5 meses e intervalo de confiança de 95% e o grupo tratado com ressecção segmentar 38% com intervalo de 27 a 48 meses, erro padrão de 5 meses, um intervalo de confiança de 95%. A comparação pelo teste de Log Rank, não paramétrico apresentou p<0,8329 e pelo teste t-Student p< 0,621 ambos não significantes. As principais complicações foram a infecção local em 5 (11,9%) pacientes e a fístula orocutânea em 4 (9,5%). Houve uma fratura da placa de titânio, dois pacientes evoluíram com osteorradionecrose e nove com disfunção da articulação têmporo-mandibular. CONCLUSÕES: Não houve diferenças estatisticamente significantes entre os grupos de ressecção marginal e segmentar nos critérios analisados. Portanto, a conservação do ramo ascendente da mandíbula, em lesões que não apresentem envolvimento mandibular, mesmo avançadas, não aumenta o índice de recidiva. / INTRODUCTION: For several decades, resection of the ascending ramus of the mandible was considered to be mandatory for the treatment of tonsil and retromolar trigone tumours, independent from the damage degree of the mandibular bone, causing considerable functional and aesthetic deficit, many times with irreparable quality of life loss, sometimes unnecessary. Thus, marginal resection of the mandibular bone appeared as a feasible alternative treatment, since maintaining a segment of the mandibular ramus in lesions without bone involvement did not increase the recurrence indexes or compromise the principles of oncological radicalness. OBJECTIVES AND METHODS: Through a retrospective study from October 1994 to December 2001, 42 patients with advanced retromolar and tonsil tumors were compared, 20 undergoing marginal resection of the mandibular bone and 22 undergoing segmental resection of the ascending ramus of the mandible, with regard complications, injury originated from the procedure, locoregional recurrence and survival. RESULTS: From the 20 patients undergoing to marginal mandibulectomy, assessed for a period of 09 to 60 months, seven (35%) patients died of the disease, with a minimal survival of 9 months: 3 due to local recurrence, 3 due to regional recurrence and 1 due to local and regional recurrence. One patient died in the immediate postoperative period. When assessing the surgical part, all the margins were found to be free and in two patients, were considered to be of little depth, one of them being found in one of the patients that died from local recurrence. There was lymph nodal dissemination in 15 patients, with capsular rupture in 4, of which two presented regional recurrence. The locoregional control was obtained in 63% of the patients. From the 22 patients undergoing to segmental resection of the mandibular bone, with a follow-up varying from 14 to 60 months, 8 (36.4%) died of the disease, with a minimum survival of 9 months, 5 due to local recurrence and 3 due to distant recurrence. One patient died in the immediate postoperative period. The surgical margins were considered free from disease in 20 patients and, in three they were too small, one patient died of local recurrence. There was lymph nodal dissemination in 12 patients, and 7 presented capsular rupture. The locoregional control was obtained in 61% of the patients. In the survival analysis curve, by the Kaplan-Meier method, the group submitted to marginal mandibulectomy presented with a rate of 42%, with an interval of 31 to 52 months, standard error of 5 months and confidence interval of 95%, and the group submitted to segmentary resection, 38% with an interval of 27 to 48 months, a standard error of 5 months, and a confidence interval of 95%. The comparison using the non-parametric Log-Rank test presented p<0.8326 and the t-Student test p< 0.621, both not statistically significant. The main complications were local infection in 5 (11.9%) patients and oro-cutaneous fistula in 4 (9.5%). There was one titanium plate fracture, two patients developed osteoradionecrosis and nine temporo-mandibular dysfunction. CONCLUSIONS: There weren t statistically significant differences between the marginal and the segmental resection groups in relation to the analyzed criteria. So, the preservation of the ascending ramus of the mandible, in lesions which do not present mandible commitment, even if not advanced, doesn t increase the recurrence rate.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-01-29T08:31:16Z No. of bitstreams: 2 Dissertação - Carlos Henrique Pereira - 2014.pdf: 3850945 bytes, checksum: 985b041e7a863070b3fa0562180a40f3 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-01-29T08:33:08Z (GMT) No. of bitstreams: 2 Dissertação - Carlos Henrique Pereira - 2014.pdf: 3850945 bytes, checksum: 985b041e7a863070b3fa0562180a40f3 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-01-29T08:33:08Z (GMT). No. of bitstreams: 2 Dissertação - Carlos Henrique Pereira - 2014.pdf: 3850945 bytes, checksum: 985b041e7a863070b3fa0562180a40f3 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-07-07 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Objective: Investigate, on a comparative basis, the expression of the adhesion molecules E-cadherin (E-cad), β-catenin (β-cat) and the proliferation index (Ki-67) at the invasive tumor front (ITF) in Squamous Cell Carcinoma (SCC) and basaloid squamous cell carcinoma (BSCC). Material and Methods: Thirty-five SCC and 16 BSCC cases were evaluated by immunohistochemistry. Clinico-pathological data and survival data were evaluated and compared. Results: There was a low expression of E-cad in the cytoplasmic membrane (p = 0.50) as well as in the nucleus (p = 0.31) for both SCC and BSCC. A high expression of E-cad was seen in the cytoplasm for the SCC group (80%) when compared to the BSCC group (25%) (p<0.01). The expression of β-cat in the cytoplasmic membrane (p = 0.28) and in the cytoplasm (p = 0.44) was low in both SCC and BSCC groups. Both types of carcinoma presented low expressions of β-cat in the nucleus (p = 0.03). The Ki-67 expression was low irrespective of tumor variant. The high expression of E-cad in the cytoplasm was associated with T3/T4 tumors (p = 0.04) in the SCC group and there was no significant association of E-cad, β-cat, Ki-67 with the other clinical variables. In terms of disease-free survival and overall survival, there were no significant differences between SCC and BSCC. Conclusion: The E-cad-β-cat system was found to be dysregulated in both oral SCC and oral BSCC. The Ki-67 cell proliferation index was extremely low in the cases investigated and consequently had no prognostic value. / Objetivo: Investigar comparativamente a expressão das moléculas de adesão E-caderina (E-cad), β-catenina (β-cat) e o índice de proliferação (Ki-67) em Carcinoma Espinocelular (CEC) e Carcinoma escamoso basalóide (CEB). Material e Métodos: Nesse estudo, foram selecionados 35 casos de CEC e 16 casos de CEB e investigados por meio de técnica imuno-histoquímica. A associação das variáveis clinico-patológicas e dados de sobrevida foram avaliados. Resultados: Ambos os grupos tiveram baixa expressão de E-cad em membrana citoplasmática (p=0,50) e núcleo (p=0,31), essa associação não foi estatisticamente significante entre os grupos. Alta expressão de E-cad no citoplasma foi notada no grupo CEC (80%) quando comparado ao grupo CEB (25%), p<0,01. A expressão de β-cat em membrana citoplasmática (p=0,28) e citoplasma (p=0,44) foram baixos em CEC e CEB. Ambos os grupos tiveram baixa expressão de β-cat em núcleo, p=0,03. Não houve diferença estatisticamente significante quanto à expressão de Ki-67 entre os grupos. Alta expressão de E-cad em citoplasma foi associada a tumores T3/T4 (p=0,04) no grupo CEC. Pacientes estilistas apresentaram baixa expressão de β-cat em membrana citoplasmática (p=0,05). Não houve associação entre a expressão de E-cad, β-cat e Ki-67 com as demais variáveis clínicas dos grupos. A sobrevida livre de doença e a sobrevida global não foram estatisticamente significantes na associação entre os grupos CEC e CEB. Conclusão: O sistema E-cad-β-cat encontra-se desregulado tanto nos de CEC quanto nos de CEB de cavidade oral, levando as células epiteliais a perderem o seu fenótipo e promovendo a invasão e progressão tumoral. O índice de Ki-67 foi extremamente baixo não tendo valor prognóstico nos casos avaliados.

Les carcinomes épidermoïdes des voies aéro-digestives supérieures (VADS) se situent au sixième rang des cancers les plus fréquents dans le monde. Ces tumeurs sont liés à deux facteurs de risque : l’intoxication éthylo-tabagique (80% des cas) et l’infection de l’épithélium des VADS par les papillomavirus humain (HPV) à haut risque oncogène (20% des cas). Ces derniers définissent une sous-population de patients de meilleur pronostic. Une des hypothèses actuellement étudiées, afin d’expliquer la survie améliorée des patients HPV positifs, serait une hypoxie moindre dans ces tumeurs. En effet, les tumeurs des VADS sont fréquemment hypoxiques, et l’hypoxie intratumorale est un facteur de mauvais pronostic. Dans une première partie de cette thèse, nous avons entrepris une caractérisation moléculaire de l’hypoxie intratumorale dans les tumeurs humaines oropharyngées en fonction du statut HPV. Il apparaît que les tumeurs HPV positives présentent un statut hypoxique moindre comparées aux tumeurs HPV négatives. Ces tumeurs se caractérisent également par une abondante vascularisation intratumorale, qui pourrait être à l’origine de ce statut hypoxique moindre. Dans une deuxième partie, nous avons étudié l’adaptation à l’hypoxie de la lignée cellulaire HPV négative SQ20B et la lignée cellulaire HPV positive SCC90. De plus, des modèles de xénogreffes ont été établis à partir de ces mêmes lignées cellulaires et ont été analysés du point de vue de l’hypoxie intratumorale. De façon comparable aux tumeurs HPV positives, les xénogreffes obtenus à partir de la lignée SCC90 montre un statut hypoxique réduit comparés aux xénogreffes SQ20B. Les deux lignées cellulaires s’adaptent également différemment en hypoxie in vitro. La réponse à l’hypoxie dans la lignée SCC90 semble plus dynamique. En effet, la lignée SCC90 tente de s’adapter et de répondre à cet environnement hypoxique en induisant de fort niveau d’expression de gènes comparée à la lignée SQ20B. / Head and neck squamous cell carcinoma (HNSCC) represents the sixth most common malignancy worldwide. The major risk factors for HNSCC identified are tobacco use and alcohol consumption (80% of all HNSCC), which seem to have a synergistic effect. A subgroup of HNSCCs (20% of cases), particularly those of the oropharynx, is caused by infection with high-risk types of human papillomavirus (HPV). Human papillomavirus HPV-related oropharyngeal squamous cell carcinoma defines a distinct clinical subgroup of head and neck cancer patients with improved prognosis. Currently, one of the several hypothesis studied to account for their improved survival outcomes could be a distinct hypoxia status compared to their HPV-negative counterpart. Indeed, tumour hypoxia is common in solid tumours including head and neck tumours, and hypoxia is a well-known poor prognosis factor. In first part of this thesis, we have performed a molecular characterisation of tumor hypoxia on cohort of oropharyngeal tumours according to HPV status of the patients. The results support the hypothesis that HPV-related tumours display a lesser hypoxia status compared to HPV-negative oropharyngeal tumours. These HPV-related tumours also characterize by an abundant tumour vascularisation, which could be responsible for a lesser hypoxia status. In a second part, we have studied the ability of the adaptation to hypoxia of the HPV-positive SCC90 cell line and HPV-negative SQ20B cell line. Furthermore, HPV-positive and HPV-negative HNSCC xenograft models have been established and have been analysed about tumor hypoxia. Similar to HPV-related HNSCC, tumours-derived HPV positive cell lines display a reduced hypoxic status compared to tumours-derived HPV negative cell lines. The two cell lines adapt also differently to in vitro hypoxia. In the HPV-positive cell line, the hypoxia response pathways could be more dynamics. Indeed, SCC90 cell lines attempt to adapt and to reply to hypoxic environment inducing highly expression of all of the hypoxia related genes compared to SQ20B cell lines.

published_or_final_version / abstract / Molecular Biology / Master / Master of Philosophy