Matrix metalloproteinases (MMP) are a family of proteolytic enzymes; these enzymes participate in tumour angiogenesis, processes of tumour growth and development of metastases. Therefore, it is important to evaluate the changes of expression of these enzymes in cancer patients. The aim of this study – to assess the expression of MMP genes and single nucleotide polymorphism in promoter sequences of MMP genes in breast and prostate cancer and evaluate parameters investigated regarding prognosis. The perspective study included 88 female patients with breast cancer and 82 patients with prostate cancer. MMP-9 and MMP-11 gene expression was examined and MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T) polymorphism was evaluated in the blood of breast cancer patients; MT1-MMP, MMP-1, MMP-2, MMP-3, MMP-9, MMP-11, MMP-13 gene expression was examined in the tumours. MMP-9, MMP-11 gene expression and MMP-9 (-1562 C/T) polymorphism were examined in the blood of prostate cancer patients. The results of the study show that MMP gene expression is related to clinical and pathological characteristics of breast and prostate cancer patients. MMP-3 (-1171 5A/6A) polymorphism is a prognostic factor of survival of breast cancer patients; while MMP-11 gene expression in blood is a prognostic factor of survival of prostate cancer patients. / Matrikso metaloproteinazės (MMP) yra šeima proteolitinių fermentų, dalyvaujančių naviko angiogenezėje, jo augimo ir metastazavimo procesuose. Siekiant įvertinti sergančiųjų vėžiu prognozę, svarbu nustatyti šių fermentų raiškos pokyčius. Šio disertacinio darbo tikslas – nustatyti MMP genų raišką ir vieno nukleotido polimorfizmą MMP genų promotorinėse sekose sergant krūties ir priešinės liaukos vėžiu bei įvertinti tirtuosius parametrus prognoziniu aspektu. Į perspektyvinį tyrimą buvo įtrauktos 88 pacientės, sergančios krūties vėžiu ir 82 pacientai, sergantys priešinės liaukos vėžiu. Sergančių krūties vėžiu pacienčių kraujyje buvo tirta MMP-9 ir MMP-11 raiška bei MMP-3 (-1171 5A/6A) ir MMP-9 (-1562 C/T) polimorfizmas, o navikuose – MT1-MMP, MMP-1, MMP-2, MMP-3, MMP-9, MMP-11 ir MMP-13 genų raiška. Sergančiųjų priešinės liaukos vėžiu kraujyje tirta MMP-9 ir MMP-11 genų raiška bei MMP-9 (-1562 C/T) polimorfizmas. Tyrimo rezultatai parodė, kad tirtų MMP genų raiška tiek krūties, tiek priešinės liaukos vėžio atvejais yra susijusi su pacientų klinikinėmis-patologinėmis charakteristikomis. Sergančiųjų krūties vėžiu išgyvenamumo prognozinis veiksnys yra MMP-3 (-1171 5A/6A) polimorfizmas, o sergančiųjų priešinės liaukos vėžiu – MMP-11 geno raiška kraujyje.

Matrikso metaloproteinazės (MMP) yra šeima proteolitinių fermentų, dalyvaujančių naviko angiogenezėje, jo augimo ir metastazavimo procesuose. Siekiant įvertinti sergančiųjų vėžiu prognozę, svarbu nustatyti šių fermentų raiškos pokyčius. Šio disertacinio darbo tikslas – nustatyti MMP genų raišką ir vieno nukleotido polimorfizmą MMP genų promotorinėse sekose sergant krūties ir priešinės liaukos vėžiu bei įvertinti tirtuosius parametrus prognoziniu aspektu. Į perspektyvinį tyrimą buvo įtrauktos 88 pacientės, sergančios krūties vėžiu ir 82 pacientai, sergantys priešinės liaukos vėžiu. Sergančių krūties vėžiu pacienčių kraujyje buvo tirta MMP-9 ir MMP-11 raiška bei MMP-3 (-1171 5A/6A) ir MMP-9 (-1562 C/T) polimorfizmas, o navikuose – MT1-MMP, MMP-1, MMP-2, MMP-3, MMP-9, MMP-11 ir MMP-13 genų raiška. Sergančiųjų priešinės liaukos vėžiu kraujyje tirta MMP-9 ir MMP-11 genų raiška bei MMP-9 (-1562 C/T) polimorfizmas. Tyrimo rezultatai parodė, kad tirtų MMP genų raiška tiek krūties, tiek priešinės liaukos vėžio atvejais yra susijusi su pacientų klinikinėmis-patologinėmis charakteristikomis. Sergančiųjų krūties vėžiu išgyvenamumo prognozinis veiksnys yra MMP-3 (-1171 5A/6A) polimorfizmas, o sergančiųjų priešinės liaukos vėžiu – MMP-11 geno raiška kraujyje. / Matrix metalloproteinases (MMP) are a family of proteolytic enzymes; these enzymes participate in tumour angiogenesis, processes of tumour growth and development of metastases. Therefore, it is important to evaluate the changes of expression of these enzymes in cancer patients. The aim of this study – to assess the expression of MMP genes and single nucleotide polymorphism in promoter sequences of MMP genes in breast and prostate cancer and evaluate parameters investigated regarding prognosis. The perspective study included 88 female patients with breast cancer and 82 patients with prostate cancer. MMP-9 and MMP-11 gene expression was examined and MMP-3 (-1171 5A/6A), MMP-9 (-1562 C/T) polymorphism was evaluated in the blood of breast cancer patients; MT1-MMP, MMP-1, MMP-2, MMP-3, MMP-9, MMP-11, MMP-13 gene expression was examined in the tumours. MMP-9, MMP-11 gene expression and MMP-9 (-1562 C/T) polymorphism were examined in the blood of prostate cancer patients. The results of the study show that MMP gene expression is related to clinical and pathological characteristics of breast and prostate cancer patients. MMP-3 (-1171 5A/6A) polymorphism is a prognostic factor of survival of breast cancer patients; while MMP-11 gene expression in blood is a prognostic factor of survival of prostate cancer patients.

Diss. (sammanfattning) Linköping : Linköpings universitet, 2008. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 6 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

Resumo: A proposta desta pesquisa foi avaliar a capacidade de diferentes concentrações de endotoxinas (LPS) de Escherichia coli induzir secreção de MMP-3 e MMP-8 por macrófagos in vitro, em diferentes períodos de tempo. Para tanto, monócitos humanos da linhagem U937, diferenciados em macrófagos maduros com phorbol-12-myristate-13-acetate (PMA), foram ativados com 0 (controle), 5, 10 e 20 EU/mL e a produção de MMP- 3 e MMP-8 foi avaliada em três períodos de tempo (24, 48 e 72 h) por ensaios imunoenzimáticos (ELISA). Os resultados foram analisados estatisticamente (ANOVA e teste de Tukey, 5%). Pode-se verificar que houve significativo aumento da produção de MMP-3 (p<0,05) e redução da produção de MMP-8 após ativação com diferentes concentrações de LPS em relação ao controle, nos períodos de 24, 48 e 72 h. Em praticamente todos os grupos (incluindo os controles), pode-se verificar que a produção de MMP-3 ou MMP-8 aumentou significativamente em relação ao aumento do tempo. Pode-se concluir que: a) diferentes concentrações de endotoxinas apresentaram capacidade de induzir aumento significativo da produção de MMP-3 nos períodos de 24, 48 e 72 h, independente da concentração utilizada, sugerindo que esta ativação é dependente da presença de endotoxina e não da dose; b) a presença de diferentes concentrações de endotoxinas induziu significativa redução da produção de MMP-8 em relação ao controle, em todos os períodos de tempo avaliados. / Abstract: The aim of this study was to evaluate the capacity of different concentrations of endotoxins (LPS) of Escherichia coli to induce secretion of MMP-3 and MMP-8 in macrophages in vitro, in the periods of 24, 48 and 72 h. For this purpose, human monocytes line U937 were differentiated into mature macrophages with phorbol-12-myristate-13- acetate (PMA) and were activated with different concentrations of endotoxins: 0 (control), 5, 10 and 20 EU/ml and the production of MMP-3 and MMP-8 were evaluated at three periods of time (24, 48 e 72 h) by immunoenzymatic assays (ELISA). The results were analyzed statistically by ANOVA and Tukey's test, 5%. Significant increase in the production of MMP-3 (p<0.05) after the activation with different concentrations of LPS in relation to the control at the periods of 24, 48 and 72 h were observed. Regarding MMP-8, significant reduction in the production after the activation with different concentrations of endotoxins in relation to the control was observed at all the periods evaluated. Basically in all groups (including the controls) the production of MMP-3 and MMP-8 increased significantly in relation to the time. It could be concluded that: a) different concentrations of endotoxins were able to induce significant increase in the production of MMP-3 at the periods of 24, 48 and 72 h, independently of the concentration used, suggesting that this activation is dependent of the presence of the endotoxin and dosis-independent; b) the presence of different concentrations of endotoxins induced significant reduction in the production of MMP-8 in relation to the control in all the periods of evaluation. / Orientador: Antonio Olavo Cardoso Jorge / Coorientador: Luciane Dias de Oliveira / Banca: Cristiane Yumi Koga Ito / Banca: Mariella Vieira Pereira Leão / Mestre

Prostate cancer micrometastasis allows cancer cells to vacate their original tumor sites and migrate to distant parts of the body via the bloodstream, lymphatic system, or by direct extension. Cells synthesize and secrete matrix metalloproteinases (MMPs) that degrade proteins of the surrounding extracellular matrix (ECM); thus allowing them to escape into the lymphatic or circulatory systems to invade other tissues. Transforming growth factor β (TGF-β) induces the migration and invasion of cancer cells and the expression of matrix metalloproteinases (MMPs), specifically MMP-2, and -9 in several malignancies. In this study, we examined the role of MMP-7, a known activator of MMP-2 and MMP-9, in TGF-β signaling in cell proliferation, migration, and invasion in prostate cancer cells. Basal expression levels of MMP7 mRNA, protein, and secreted protein were determined using RT-PCR, western blot analysis, and ELISA, respectively. Our data show that MMP7 mRNA and proteins were differentially expressed in several cell line models representing different stages of prostate cancer. TGF-β1 induces MMP-7 gene expression and protein levels 24 and 48 hours after treatment in PC3 cells. Our data also show that TGF-β induces cell migration and invasion in PC3 and E006AA cells; however, the selective knockdown of MMP7 expression using siRNA resulted in a significant decrease in control and TGFβ-induced cell migration and invasion in both PC3 and E006AA cells. MMP-7 knockdown also caused significant reduction in cell proliferation in PC3 cells. Our data suggest that MMP7 is essential for cell migration and invasion in prostate cancer cells indicating that it may be required for TGFβ-induced cancer metastases.

Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro / O objetivo deste trabalho foi examinar o impacto da movimentação dentária de dentes periodontalmente comprometidos no volume do fluido gengival crevicular (FGC) e nos níveis de expressão das metaloproteinases de matriz (MMPs) -1, -2, -3, -7, -8, -12 e -13 no FGC. Dez pacientes periodontalmente controlados (8 do sexo feminino e 2 do sexo masculino, média de idade de 46,2 10,4 anos) com incisivos projetados labialmente foram submetidos a tratamento ortodôntico. Uma arcada dentária foi submetida a movimentação ortodôntica e a arcada oposta foi usada como controle. Amostras de FGC foram coletadas da face lingual de dois incisivos do lado movimento e dois do lado controle uma semana antes da ativação ortodôntica (-7d), imediatamente após a ativação ortodôntica, e após 1 h, 24 h, e 7, 14 e 21 dias. A coleta do FGC foi feita utilizando-se tiras de papel absorvente e o volume foi calculado através do uso do Periotron. Todos os pacientes receberam orientações de higiene bucal e um kit contendo escova de dente, pasta de dente e bochecho de Chlorexidina 0,12% para ser usado durante todo o experimento. A técnica da multianálise imunoenzimática com microesferas foi usada para medir as MMPs no FGC. Os dados foram analisados utilizando-se os testes estatísticos Friedman e Mann-Whitney. Não foram encontradas diferenças estatisticamente significativas no volume do FGC. Em relação aos níveis de MMPs, a única diferença estatisticamente significativa encontrada no decorrer do tempo foi nos níveis de MMP-1 no grupo movimento (p<0,05). Quando os dois grupos foram comparados após a ativação, a única diferença estatisticamente significativa encontrada foi nos níveis de MMP-12 24 horas após a ativação (p<0,05). Estes achados sugerem que o volume de FGC não sofre alteração relacionada ao movimento dentário ortodôntico e que o movimento ortodôntico de dentes periodontalmente comprometidos não resultou em mudanças significativas nos níveis de MMPs no FGC. / The aim of this study was to evaluate whether orthodontic tooth movement of periodontally compromised teeth would result in detectable changes in gingival crevicular fluid (GCF) volume and in the levels of matrix metalloproteinases (MMP) -1, -2, -3, -7, -8, -12 and -13 in the GCF. Ten controlled periodontitis subjects (8 females and 2 males, mean age of 46.2 10.4 years) with flared incisors were submitted to orthodontic treatment. One dental arch was subjected to orthodontic movement, and teeth in the opposite arch were used as controls. GCF samples were collected from the lingual sites of two test and two control incisors one week before orthodontic activation (-7d), immediately after orthodontic activation, and after 1 h, 24 h, and 7, 14 and 21 days. Filter paper strips were used in combination with a Periotron for GCF volume measurements. All patients received hygiene instructions and a kit containing toothbrush, fluoride dentifrice and gluconate chlorexidine 0.12% mouthwash to be used during all experiment. Multiplexed bead immunoassay was used to measure MMPs in GCF. Data were analyzed using Friedman and Mann-Whitney statistical tests. There were no statistically significant differences over time in the GCF volume for any of the two groups. Regarding the MMPs levels, the only significant change found over time was in the levels of MMP-1 in the movement group (p<0.05). When the two groups were compared after activation, the only statistically significant difference found was in levels of MMP-12 24 h after activation (p<0.05). This findings suggested that the GCF volume could not be associated to orthodontic movement and that the orthodontic movement of periodontally compromised teeth did not result in significant changes in the GCF levels of MMPs.

Made available in DSpace on 2014-06-11T19:27:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-07-23Bitstream added on 2014-06-13T18:31:25Z : No. of bitstreams: 1 vilela_pgf_me_sjc.pdf: 389414 bytes, checksum: e94b74337996f5291f23f9666ba4f093 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A proposta desta pesquisa foi avaliar a capacidade de diferentes concentrações de endotoxinas (LPS) de Escherichia coli induzir secreção de MMP-3 e MMP-8 por macrófagos in vitro, em diferentes períodos de tempo. Para tanto, monócitos humanos da linhagem U937, diferenciados em macrófagos maduros com phorbol-12-myristate-13-acetate (PMA), foram ativados com 0 (controle), 5, 10 e 20 EU/mL e a produção de MMP- 3 e MMP-8 foi avaliada em três períodos de tempo (24, 48 e 72 h) por ensaios imunoenzimáticos (ELISA). Os resultados foram analisados estatisticamente (ANOVA e teste de Tukey, 5%). Pode-se verificar que houve significativo aumento da produção de MMP-3 (p<0,05) e redução da produção de MMP-8 após ativação com diferentes concentrações de LPS em relação ao controle, nos períodos de 24, 48 e 72 h. Em praticamente todos os grupos (incluindo os controles), pode-se verificar que a produção de MMP-3 ou MMP-8 aumentou significativamente em relação ao aumento do tempo. Pode-se concluir que: a) diferentes concentrações de endotoxinas apresentaram capacidade de induzir aumento significativo da produção de MMP-3 nos períodos de 24, 48 e 72 h, independente da concentração utilizada, sugerindo que esta ativação é dependente da presença de endotoxina e não da dose; b) a presença de diferentes concentrações de endotoxinas induziu significativa redução da produção de MMP-8 em relação ao controle, em todos os períodos de tempo avaliados. / The aim of this study was to evaluate the capacity of different concentrations of endotoxins (LPS) of Escherichia coli to induce secretion of MMP-3 and MMP-8 in macrophages in vitro, in the periods of 24, 48 and 72 h. For this purpose, human monocytes line U937 were differentiated into mature macrophages with phorbol-12-myristate-13- acetate (PMA) and were activated with different concentrations of endotoxins: 0 (control), 5, 10 and 20 EU/ml and the production of MMP-3 and MMP-8 were evaluated at three periods of time (24, 48 e 72 h) by immunoenzymatic assays (ELISA). The results were analyzed statistically by ANOVA and Tukey’s test, 5%. Significant increase in the production of MMP-3 (p<0.05) after the activation with different concentrations of LPS in relation to the control at the periods of 24, 48 and 72 h were observed. Regarding MMP-8, significant reduction in the production after the activation with different concentrations of endotoxins in relation to the control was observed at all the periods evaluated. Basically in all groups (including the controls) the production of MMP-3 and MMP-8 increased significantly in relation to the time. It could be concluded that: a) different concentrations of endotoxins were able to induce significant increase in the production of MMP-3 at the periods of 24, 48 and 72 h, independently of the concentration used, suggesting that this activation is dependent of the presence of the endotoxin and dosis-independent; b) the presence of different concentrations of endotoxins induced significant reduction in the production of MMP-8 in relation to the control in all the periods of evaluation.