Tumour necrosis factor stimulated gene 6 (TSG-6) is a protein present in a range of tissues and is produced by a wide range of cell types in response to a number of inflammatory stimuli, where this protein is thought to mediate protection against excessive inflammation. TSG-6 is expressed in response to inflammation and has been implicated as an endogenous protector of tissues, e.g. in the context of inflammatory arthritis. TSG-6 has also been shown to reduce inflammatory damage in animal models of both myocardial infarction and corneal injury. Our earlier studies demonstrated that TSG-6 is a potent inhibitor of neutrophil migration, which likely contributes to these protective activities. Here we investigated the effect of TSG-6 on CXCL8-mediated pro-inflammatory processes. The interaction of TSG-6 with CXCL8, and how this influences the binding of CXCL8 to heparin, was investigated using solid-phase assays and surface plasmon resonance (SPR). The ability of this interaction to inhibit the interaction between CXCL8 and one of its receptors CXCR2 was investigated using murine pre-B cells expressing this receptor, in flow cytometry experiments. The effects of TSG-6 on CXCL8's pro-inflammatory activities were assessed using a neutrophil cell line (differentiated HL60 cells) in a trans-endothelial migration assay and gelatin zymography to measure secretion of MMPs by the endothelial cell (EC) line EA.hy 926. TSG-6 expression in EA.hy 926 and HL-60 cells was assessed using qRT-PCR, immunofluorescence and western blotting of cell lysates and culture media. We have shown that TSG-6 binds to CXCL8 via its Link module domain (Link_TSG6) and inhibits the interaction of CXCL8 with heparin. Analysis of culture media from EA.hy 926 cells revealed that both full-length TSG-6 and Link_TSG6 abolished CXCL8-mediated up-regulation of MMP-2 secretion. In transmigration assays, TSG-6 and Link_TSG6 were found to inhibit CXCL8-induced migration of neutrophils across an EC monolayer and also inhibited the interaction between CXCL8 and CXCR2; this effect was enhanced in mutants of Link_TSG6 with reduced heparin-binding functions. Very limited TSG-6 expression was observed in EA.hy 926 and HL-60 cells, where stimulation with pro-inflammatory mediators had little effect on expression. Here we have shown that TSG-6 binds directly to CXCL8 and inhibits its interaction with heparin, its interaction with CXCR2 and its enhancement of MMP-2 secretion by ECs; these effects are mediated via the Link module of TSG-6. Furthermore, Link_TSG6 inhibits trans-endothelial migration of neutrophils in a dose dependent manner; this could be due in part to reduced association of CXCL8 with EC glycosaminoglycans or with its receptors on neutrophils, thereby limiting its pro-migratory activity. Inhibition of MMP production by ECs could also limit neutrophil trans-migration as well as tissue damage and angiogenesis. Thus, the modulation of CXCL8 activity represents one way in which TSG-6 might protect tissues from the damaging effects of inflammation.

Elevated deoxycholic acid (DCA) and mutations in the Adenomatous Polyposis Coli (APC) tumor suppressor gene have been associated with increased risk of colorectal cancer (CRC). Chronic inflammation has also been associated with increased risk of CRC. It is unclear if DCA mediates inflammation in the normal or transformed colonic mucosa. The status of APC was manipulated in human CRC cell lines to study the role of DCA mediated inflammation. The chemotactic cytokine, CXCL8, was used as a marker of inflammation. Addition of DCA to the HT29-parental cell line with mutant-APC increased the steady state mRNA and protein levels of CXCL8. Conversely, addition of DCA to the HT29-APC cell line with wild type-APC was protective for increased steady state RNA and protein levels of CXCL8. DCA activated transcription factors which had binding regions in the CXCL8 5’-promoter. To elucidate the mechanism of induction, the 5’-promoter of CXCL8 was investigated. DCA increased promoter-reporter activity of the CXCL8 gene in HT29-parental cell line but wild type-APC blocked this effect. Chromatin immunoprecipitation (ChIP) revealed that DCA activated transcription factors, AP-1 and NF-κB were bound to the 5’-promoter of CXCL8. The transcription factor, β-catenin, was also bound to the 5’-promoter of CXCL8. Phenotypic effects were measured. Increased CXCL8 lead to matrix metalloproteinase-2 (MMP-2) production and increased invasion by HT29-parental cells on laminin coated filters. The DCA-mediated invasion was blocked by antibody directed against CXCL8 and wild type- APC. Therefore DCA-mediated inflammation occurs in transformed colonic epithelium and increases the invasive phenotype of CRC cells by CXCL8.

Um material obturador deve apresentar boas propriedades biológicas e físico-químicas, uma vez que ficará em íntimo contato com os tecidos periapicais. Sendo assim, não deve ser irritante aos tecidos adjacentes, possibilitando, ou mesmo favorecendo, o reparo da região periapical, para isso não devem alterar o processo inflamatório. O presente estudo avaliou a citotoxicidade dos cimentos AH Plus, Sealapex, MTA Fillapex e Sealepox em neutrófilos humanos. Neutrófilos humanos cultivados na presença ou ausência de LPS foram tratados com os cimentos em diferentes diluições (1:1, 1:4, 1:8, 1:16 e 1:32) e tempo de presa (24 e 48 horas) durante 24 horas. A viabilidade celular foi analisada por citometria de fluxo, a dosagem de CXCL8 pelo método de ELISA, e Óxido Nítrico na absorvância de 540nm. Os dados foram analisados com o auxílio do programa GraphPad Prism5 por ANOVA a 1 critério seguido pelo teste de Tukey, e os valores considerados significantes quando p<0,05. O cimento AH Plus interferiu apenas na síntese de NO, estimulando-a, tendo a diluição 1:16 melhor comportamento biológico, em ambos períodos experimentais. O Sealapex diminuiu a produção de NO, sendo significante para 1:32 em 48 horas. O MTA Fillapex induziu apoptose, a produção de CXCL8 (1:4 e 1:8 em 48 horas) e diminuiu a síntese de NO (1:32 em 48 horas). Sealepox diminuiu a apoptose (1:16 e 1:32 em 24 horas) e interferiu na produção de CXCL8, diminuindo-a (1:8 em 48 horas, e 1:16 em ambos os períodos). A citotoxicidade em ordem crescente dos cimentos foram: AH Plus, Sealapex, MTA Fillapex e Sealepox. Nosso estudo concluiu que os cimentos AH Plus e Sealapex foram os menos citotóxicos, que menos interferiram na viabilidade celular e na sua função (não indução de CXCL8 e na produção de NO), tanto em 24 horas como em 48 horas. MTA Fillapex e o Sealepox, apesar de causarem mais morte celular e interferirem na produção de NO e CXCL8, seus efeitos podem ser aceitáveis, uma vez que os níveis dessas alterações são, de tal maneira discretos, não agressivos. / A filling material must have good biological and physicochemical, since you\'ll be in close contact with the periapical tissues. Thus it should not be irritating to the surrounding tissues, allowing, or even encouraging, the repair of the periapical region, for it must not alter the inflammatory process. The present study evaluated the cytotoxicity of AH Plus, Sealapex, MTA and Fillapex Sealepox in human neutrophils. Human neutrophils cultured in the presence or absence of LPS cements were treated with different dilutions (1:1, 1:4, 1:8 , 1:16, 1:32) and setting time (24 and 48 hours) for 24 hours. Cell viability was analyzed by flow cytometry, the dosage of CXCL8 by ELISA and nitric oxide in absorbance at 540 nm. Data were analyzed with the aid of GraphPad Prism5 ANOVA for the first criterion followed by Tukey test, and values considered significant when p < 0.05. The AH Plus sealer interfered only in NO synthesis , stimulating it , having a 1:16 dilution better biological behavior , in both experimental periods . The Sealapex decreased production of NO, with 1:32 significant to within 48 hours. The MTA Fillapex induced apoptosis, production of CXCL8 (1:4 to 1:8 within 48 hours) and reduced NO synthesis (1:32 in 48 hours). Sealepox decreased apoptosis (1:16 and 1:32 in 24 hours) and interfere with the production of CXCL8, reducing it (1:8 in 48 hours, and 1:16 in both periods). The cytotoxicity of the cements in ascending order were: AH Plus, Sealapex, MTA and Fillapex Sealepox. We conclude that AH Plus and Sealapex cements were less cytotoxic than less interfered with cell viability and function ( CXCL8 and noninduced NO production ) both in 24 hours and in 48 hours. MTA and Fillapex Sealepox, although most causing cell death and interferes the production of NO and CXCL8, their effects may be acceptable, since the levels of these changes are so mild, non-aggressive.

Um material obturador deve apresentar boas propriedades biológicas e físico-químicas, uma vez que ficará em íntimo contato com os tecidos periapicais. Sendo assim, não deve ser irritante aos tecidos adjacentes, possibilitando, ou mesmo favorecendo, o reparo da região periapical, para isso não devem alterar o processo inflamatório. O presente estudo avaliou a citotoxicidade dos cimentos AH Plus, Sealapex, MTA Fillapex e Sealepox em neutrófilos humanos. Neutrófilos humanos cultivados na presença ou ausência de LPS foram tratados com os cimentos em diferentes diluições (1:1, 1:4, 1:8, 1:16 e 1:32) e tempo de presa (24 e 48 horas) durante 24 horas. A viabilidade celular foi analisada por citometria de fluxo, a dosagem de CXCL8 pelo método de ELISA, e Óxido Nítrico na absorvância de 540nm. Os dados foram analisados com o auxílio do programa GraphPad Prism5 por ANOVA a 1 critério seguido pelo teste de Tukey, e os valores considerados significantes quando p<0,05. O cimento AH Plus interferiu apenas na síntese de NO, estimulando-a, tendo a diluição 1:16 melhor comportamento biológico, em ambos períodos experimentais. O Sealapex diminuiu a produção de NO, sendo significante para 1:32 em 48 horas. O MTA Fillapex induziu apoptose, a produção de CXCL8 (1:4 e 1:8 em 48 horas) e diminuiu a síntese de NO (1:32 em 48 horas). Sealepox diminuiu a apoptose (1:16 e 1:32 em 24 horas) e interferiu na produção de CXCL8, diminuindo-a (1:8 em 48 horas, e 1:16 em ambos os períodos). A citotoxicidade em ordem crescente dos cimentos foram: AH Plus, Sealapex, MTA Fillapex e Sealepox. Nosso estudo concluiu que os cimentos AH Plus e Sealapex foram os menos citotóxicos, que menos interferiram na viabilidade celular e na sua função (não indução de CXCL8 e na produção de NO), tanto em 24 horas como em 48 horas. MTA Fillapex e o Sealepox, apesar de causarem mais morte celular e interferirem na produção de NO e CXCL8, seus efeitos podem ser aceitáveis, uma vez que os níveis dessas alterações são, de tal maneira discretos, não agressivos. / A filling material must have good biological and physicochemical, since you\'ll be in close contact with the periapical tissues. Thus it should not be irritating to the surrounding tissues, allowing, or even encouraging, the repair of the periapical region, for it must not alter the inflammatory process. The present study evaluated the cytotoxicity of AH Plus, Sealapex, MTA and Fillapex Sealepox in human neutrophils. Human neutrophils cultured in the presence or absence of LPS cements were treated with different dilutions (1:1, 1:4, 1:8 , 1:16, 1:32) and setting time (24 and 48 hours) for 24 hours. Cell viability was analyzed by flow cytometry, the dosage of CXCL8 by ELISA and nitric oxide in absorbance at 540 nm. Data were analyzed with the aid of GraphPad Prism5 ANOVA for the first criterion followed by Tukey test, and values considered significant when p < 0.05. The AH Plus sealer interfered only in NO synthesis , stimulating it , having a 1:16 dilution better biological behavior , in both experimental periods . The Sealapex decreased production of NO, with 1:32 significant to within 48 hours. The MTA Fillapex induced apoptosis, production of CXCL8 (1:4 to 1:8 within 48 hours) and reduced NO synthesis (1:32 in 48 hours). Sealepox decreased apoptosis (1:16 and 1:32 in 24 hours) and interfere with the production of CXCL8, reducing it (1:8 in 48 hours, and 1:16 in both periods). The cytotoxicity of the cements in ascending order were: AH Plus, Sealapex, MTA and Fillapex Sealepox. We conclude that AH Plus and Sealapex cements were less cytotoxic than less interfered with cell viability and function ( CXCL8 and noninduced NO production ) both in 24 hours and in 48 hours. MTA and Fillapex Sealepox, although most causing cell death and interferes the production of NO and CXCL8, their effects may be acceptable, since the levels of these changes are so mild, non-aggressive.

In the first part of my dissertation we focused on the development of covalently linking compounds that bind gp120 with those that bind gp41 in order to block HIV fusion. We used griffithsin or CD4M33, that both bind to gp120, covalently linked with C-peptide C37 of gp41. The results show the linked compound Griff37 is several-fold more potent than griffithsin alone in both fusion and viral assay, making Griff37 an attractive candidate for further development as a microbicide against HIV. In the second part of my dissertation we investigate the effect of combining HIV fusion inhibitors having differing mechanisms of action. We used P2-RANTES and RANTES that both bind to the chemokine receptor CCR5 on the surface of the human cells along with C-peptides C37 and C34 that bind the viral envelop glycoprotein gp41. We found that the combination of RANTES with C37/C34 has an additive effect on fusion assay. In contrast P2-RANTES and C37/C34 shows synergy in inhibition of cell fusion. In viral assays using MAGI cells the synergy between P2-RANTES and C37/C34 is minimal. In the third part of my dissertation we focused on characterization of two examples of evolutionarily conserved unfavorable sequence motifs that affect quaternary structure. In contrast to the straightforward action of favorable sequences, these unfavorable motifs produce interactions disfavoring one outcome to indirectly promote another one. To identify such motifs, we propose and developed a statistically validated computational method combining structure and phylogeny. This approach was applied in an analysis of the alternate forms of homodimerization exhibited in the chemokine family. We identified two evolutionarily conserved sequence motifs in the CC subfamilies: a drastic two-residue deletion (ERV) and a simple point mutation (V27R). Cloned into the CXCL8 background, these two motifs were experimentally proven to confer a monomeric state. NMR analyses indicate that these variants are structured in solution and retain the chemokine fold. Structurally, the motifs retain a chemokine tertiary fold while introducing unfavorable quaternary interactions that inhibit CXCL8 dimerization.

Orientador: Claudia Aparecida Rainho / Resumo: O carcinoma mamário é o tipo de câncer mais frequente entre as mulheres, com estimativas de aumento de sua incidência mundial nas próximas décadas. O câncer de mama triplo negativo (CMTN) é um subtipo definido pela falta de receptores para os hormônios estrógeno e progesterona e sem superexpressão de HER-2. Ocorre geralmente em mulheres com menos de 50 anos e está relacionado a uma alta taxa de recorrência e mortalidade. As terapias convencionais são hormônio-dependentes ou possuem a proteína HER2 como alvo, logo o CMTN não é responsivo a essas modalidades terapêuticas. Devido a biologia deste tumor, novas abordagens são necessárias para o seu tratamento. Plantas são importantes fontes de compostos antitumorais. Os sesquiterpenos α-humuleno (HUM) e β-cariofileno (CAR) geralmente estão presentes em conjunto no óleo essencial de diversas plantas. Um número limitado de estudos foi realizado sobre os seus efeitos em células derivadas de carcinomas mamários, porém um efeito imunomodulador foi descrito para diferentes sesquiterpenos em células derivadas de cânceres humano. A hipótese desse estudo é que os efeitos in vitro dos sesquiterpenos HUM e CAR nas linhas celulares derivadas de CMTP são mediados por alterações nos níveis de expressão de citocinas produzidas pelas células tumorais. Foram selecionadas quatro linhagens celulares triplo-negativas (BT-20, BT-549, MDA-MB-231 e MDA-MB-436). Inicialmente, os efeitos isolados ou combinados do HUM e CAR foram avaliados pelo ensaio de v... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Breast cancer is the most common type of cancer amongst women. Worldwide, its incidence is estimated to increase in the coming decades. Triple negative breast cancer (TNBC) is a subtype defined by the absence of receptors for estrogen and progesterone hormones and without HER-2 overexpression. It usually occurs in women under 50 years old and it is related to a high recurrence and mortality rates. As conventional therapies are hormone-dependent or target HER2, TNBC are not responsive to these therapeutic approaches. Due to the biology of this tumor, new targets are needed for its treatment. Plants are important sources of antitumor compounds, the sesquiterpenes α-humulene (HUM) and β-caryophyllene (CAR) are usually present together in the essential oil of several plants. Few studies have been conducted on their effect in breast cancer cells, but an immunomodulatory effect of different sesquiterpenes was described on human cancer cells. The hypothesis of this study is that the in vitro effects of sesquiterpenes HUM and CAR on cell lines derived from TNBC are mediated by changes in cytokine expression levels produced by tumor cells. For this, four triple-negative cell lines were selected (BT-20, BT-549, MDA-MB-231, and MDA-MB-436). Initially, the isolated or combined effects of HUM and CAR were evaluated by the MTT cell viability assay upon different concentrations (6.25, 12.5, 25, 50, and 100µM), after 24, 48, 72, and 96 hours of exposure. Based on the effect of the sesquiterp... (Complete abstract click electronic access below) / Mestre

Pendant très longtemps, les traitements contre les cancers se sont basés sur la cytotoxicité des chimiothérapies, sur leur capacité à tuer directement les cellules malignes ou à induire leur senescence. Mais cette cytotoxicité accrue et non ciblée a également pour effet de tuer les cellules du système immunitaire du patient. Cependant, il a été montré que la radiothérapie, les anthracyclines ainsi que l’oxaliplatine étaient capables d’induire une apoptose décrite comme étant une mort cellulaire immunogène. De ce fait, les cellules tumorales mourantes agiront comme vaccin thérapeutique.La mort cellulaire immunogène se caractérise par trois grands marqueurs : un stress du réticulum endoplasmique pré-mortem qui va induire la translocation de la calréticuline de la lumière du réticulum endoplasmique vers la surface cellulaire, la libération d’ATP dans le milieu extracellulaire permettant le recrutement des cellules dendritiques et l’activation de l’inflammasome NLRP3 via le récepteur P2RX7, et enfin la libération de la protéine HMGB1 dans le milieu extracellulaire, qui va aller interagir avec TLR4 à la surface des cellules dendritiques pour stimuler leur fonction présentatrice d’antigène. La première partie de ce travail a consisté à comprendre les mécanismes moléculaires précis par lesquels l’ATP est sécrétée activement lors d’une mort cellulaire immunogène. En utilisant une combinaison de techniques impliquant des criblages pharmacologiques, des techniques de monitorage de la localisation intracellulaire de l’ATP entre autres, nous avons montré qu’après un traitement par les inducteurs de la mort immunogène, l’ATP était redistribué des lysosomes aux autolysosomes et que sa sécrétion requiert la protéine lysosomale LAMP1. Nous avons également montré qu’il existait d’autres voies de libération d’ATP telles que la voie de signalisation Rho, et également l’ouverture des hémicanaux pannexine 1 (PANX1). De façon surprenante, nous avons observé une implication de PANX1 dans la translocation de LAMP1 à la surface cellulaire. Ces résultats ont permis de comprendre un peu plus précisément les mécanismes de sécrétion d’ATP dans la mort cellulaire immunogène, mettant en évidence l’importance de l’exocytose lysosomale caspases dépendante et PANX1 dépendante.La seconde partie de ce travail s’est portée sur l’étude d’une autre caractéristique de la mort cellulaire immunogène, à savoir l’exposition de la calréticuline à la surface cellulaire. En partant du constat qu’après un traitement par la mitoxantrone, la calréticuline était relocalisée en périphérie à la fois dans les cellules humaines et les cellules de levure, il a été suggéré que la voie d’exposition de la calréticuline était conservée phylogénétiquement. Nous avons montré que les phéromones pouvaient agir comme inducteurs physiologiques de l’exposition de la calréticuline dans les cellules de levure. Un criblage d’ARN interférant et des analyses de transcriptome nous ont permis de montré que les chimiokines, en particulier CXCL8 chez l’humain (appelé également interleukine-8) et son orthologue Cxcl2 chez la souris étaient impliquées dans la translocation de la calréticuline à la surface cellulaire. En traitant les cellules cancéreuses par la mitoxantrone, nous observons une production de CXCL8 par les cellules cancéreuses humaines in vitro et de Cxcl2 par les cellules cancéreuses murines in vivo. Un « knockdown » des récepteurs pour CXCL8/Cxcl2 réduit de manière significative l’exposition de la calréticuline à la surface cellulaire. Ces résultats ont donc montré l’importance des chimiokines dans la voie d’exposition de la calréticuline.L’ensemble de ce travail a permis de comprendre plus en détails deux des trois grandes caractéristiques de la mort cellulaire immunogène. / Cytotoxic anti-neoplastic agents were considered for a long time to mediate their therapeutic effects via their capacity to directly kill malignant cells. Nevertheless, this high cytotoxicity is non-targeted and will eventually diminish immune cells. During the last years, it has been shown that radiotherapy and some anticancer agents, such as anthracyclines and oxaliplatin, can stimulate actively anti-tumor immune responses. In fact, they can induce an immunogenic type of apoptosis, which we termed immunogenic cell death (ICD). Thereby, dying cells can act as therapeutic vaccine against residual cancer cells that overcame the initial treatment.ICD is characterized by three major hallmarks: a pre-mortem stress of the endoplasmic reticulum (ER), which triggers the translocation of the ER chaperone protein called calreticulin (CRT) to the cell surface, the secretion of ATP from apoptotic cells, which acts as a signal for the recruitment of dendritic cells and for the activation of the NLRP3 inflammasome via its receptor P2RX7, and the release of HMGB1 into the extracellular space, allowing it to interact with TLR4 and thus stimulate the antigen-presenting functions of the DCs.The first part of my work focused on the precise molecular mechanisms by which ATP is actively secreted during ICD. Using a large panel of techniques, including chemical compounds screens and monitoring the subcellular localization of ATP, we showed that following treatment of various tumor cells with ICD inducers, ATP is redistributed from lysosomes to autolysosomes and the lysosomal protein LAMP1 is required for active ATP secretion. We also showed that Rho and pannexin 1 (PANX1) are indispensable for efficient ATP release in response to ICD inducers. Surprisingly, we observed an unexpected link between PANX1 and the exposure of LAMP1 at the cell surface. These results will help to understand the mechanisms necessary for ATP secretion during ICD.In the second part of this work we further studied the surface exposure of CRT during ICD. We observed that mitoxantrone (MTX), which belongs to the group of anthracyclines, can induce a peripheral relocalisation of CRT, both in human cells and yeast cells. In addition, we showed that pheromones can act as a physiological inducer of CRT translocation in yeast. Focused siRNA screening combined with transcriptome analyses revealed that human CXCL8 (also called interleukin-8) and its mouse ortholog Cxcl2 play an essential role in the translocation of CRT to the cell surface. Interestingly, MTX-treated human cancer cells displayed an elevated production of CXCL8 in vitro. These results were confirmed in vivo, with MTX treated murine tumors, which also displayed elevated Cxcl2 levels. The MTX-induced CRT exposure was significantly reduced when we performed a knockdown of CXCL8/Cxcl2 receptors. Altogether, these results showed the importance of chemokine signaling circuitries in immunogenic CRT exposure.This work allows for the detailed understanding of the mechanisms of ICD and might thus be useful for further targeted drug development.

No description available.

Head and neck squamous cell carcinoma is the 6th most common malignancyworldwide. Recently, a link between cancer and inflammation has been found. Mediatingthis relationship are the chemotactic cytokines known as chemokines. CXCL8 (Interleukin-8), a CXC ELR+ Chemokine mainly responsible for neutrophil chemoattraction, has beenimplicated in increased tumor proliferation, migration and angiogenesis. The current studytests the effects of CXCL8 on the tumor proliferation and metastasis. By genetically modifying cells to knockdown or overexpress the CXCL8 gene we tested its biological rolein head and neck cancer progression. Overexpression of CXCL8 in HN4 tumor cells withlow endogenous CXCL8 levels was found to increase tumor growth, as judged by cellcounting and MTT assays. Conversely, RNAi-mediated knockdown of CXCL8 expressionin HN12 cells, which express high levels of this chemokine, resulted in a decrease inproliferation. Similarly, overexpression of CXCL8 enhanced migration of HN4 cells invitro, while knockdown inhibited HN12 cell migration and invasion through a basementmembrane substitute. Taken together, these findings support the hypothesis that CXCL8affects multiple processes involved in head and neck cancer tumor progression. The datasuggest that CXCL8 is a potential therapeutic target for head and neck, and other, cancers.

A Interleucina 8 (IL-8) é uma quimiocina CXC angiogênica que tem papel importante no desenvolvimento e progressão de vários tumores malignos, incluindo o câncer de próstata (CaP). O polimorfismo de nucleotídeo único (SNP) -251 T/A da região promotora do gene da IL-8, relativo ao local de início da transcrição deste gene, está associado com a produção desta citocina. O efeito da IL-8 é mediado através de dois receptores de alta afinidade, CXCR1 e CXCR2. O presente estudo investigou a influência da variação dos genes IL-8 e CXCR2 na susceptibilidade e nas características clinicopatológicas do CaP em um grupo de brasileiros. Duzentos e um pacientes e 185 controles saudáveis foram selecionados neste estudo casocontrole. Amostras de sangue foram coletadas para extração de DNA; a tipagem da IL-8 -251 T/A e CXCR2 +1208 C/T foi realizada através da reação em cadeia da polimerase com sequência específica de primers (PCR-SSP), seguida pela eletroforese em gel de agarose. O risco associado entre os genótipos, a susceptibilidade do CaP e as características do tumor, foi estimado pelo odds ratio (OR), com intervalo de confiança de 95%, usando análise de regressão logística e ajustando para idade ao diagnóstico. Encontramos uma associação estatisticamente significativa entre o genótipo heterozigoto CT do gene CXCR2 +1208 e CaP. Este genótipo foi significativamente menos frequente em pacientes com estádio clínico T3-T4 comparado com T1-T2 (56.7% versus 80.5%). Nossos achados sugerem que os portadores do genótipo CT CXCR2 +1208 tiveram um efeito protetor para estádio avançado de CaP (CT versus CC: OR ajustado = 0.25; P = 0.02). Não foi encontrada associação significativa entre o polimorfismo -251 T/A da IL-8 e os parâmetros clinicopatológicos do CaP. Estes resultados indicam que o genótipo CT do CXCR2 +1208 é menos frequente em estádios avançado de CaP, sugerindo que este receptor de quimiocina tenha um papel na patogênese desta doença. / Interleukin-8 (IL-8) is an angiogenic CXC chemokine that plays an important role in both the development and progression of several human malignancies including prostate cancer (PC). A single nucleotide polymorphism (SNP) at -251 upstream of the transcriptional start site of the IL-8 gene has been shown to influence its production. The effects of IL-8 are mediated by two highly related chemokine receptors, CXCR1 and CXCR2. The present study investigated the influence of the IL-8 and CXCR2 gene variation on susceptibility and clinicopathological characteristics of PC in a group of Brazilian subjects. Two hundred and one patients and 185 healthy controls were enrolled in a case-control study. Blood was collected for DNA extraction; typing of IL-8 -251 T/A and CXCR2 +1208 C/T genes was performed by polymerase chain reaction with sequence-specific primers (PCR-SSP), followed by agarose gel electrophoresis. Risk association between the genotypes, PC susceptibility and tumor characteristics was estimated by odds ratio (OR) and 95% confidence intervals (95% CI) using logistic regression analysis, after adjusting for age at diagnosis. A significant association was found between the heterozygous CXCR2 +1208 CT genotype and PC. The CXCR2 +1208 CT genotype was significantly less frequent in patients with clinical stage T3-T4 compared to T1-T2 (56.7 versus 80.5%). Our findings suggest that carriers of the CXCR2 +1208 CT genotype had a protective effect for advanced PC (CT versus CC: adjusted OR = 0.25; P = 0.02). No association was observed between the SNP for IL-8 -251 T/A and clinicopathological parameters of PC. These results indicated that the CXCR2 +1208 CT genotype is less frequent in advanced stages of PC, suggesting that this chemokine receptor plays a role in the pathogenesis of this disease.