Testicular germ cell cancer (TGCC) has been increasing in incidence over recent decades, and is currently the most common malignancy amongst young men resulting in significant morbidity. These tumours are believed to arise from premalignant germ cell neoplasia in situ (GCNIS) cells, which originate from the aberrant germ cell differentiation from gonocyte to spermatogonia during fetal/early postnatal life. GCNIS cells remain dormant in the testis until puberty when they are activated to become tumours. Therefore, GCNIS cells remain in a pre-invasive stage during early childhood and early adulthood prior to the development of a seminoma or non-seminoma TGCC. GCNIS cells are phenotypically similar to gonocytes with expression of stem cell/early germ cell markers including OCT4, PLAP and LIN28. Furthermore, proteins which are expressed in more mature germ cells (spermatogonia) such as MAGE-A4 have also been shown to be expressed in GCNIS cells and these studies have indicated that GCNIS cells are a heterogeneous population in terms of protein expression profile. The relationship between the protein expression profile of individual GCNIS cells populations and their oncogenic potential has not been fully explored. GCNIS cells are located in the seminiferous tubules supported by somatic Sertoli cells. These cells have been previously reported to exhibit an immature protein expression profile in GCNIS tubules from patients with testis cancer, suggesting that the germ stem cell niche in GCNIS tubules resembles that of a fetal one. Associations between Sertoli cell maturation and GCNIS progression into tumour formation has not been fully investigated. Oncogenes are key players in the regulation of oncogenic potential of cancer cells. Gankyrin is an oncogene that has been shown to down-regulate OCT4, and interact with MAGE-A4 in hepatocellular carcinoma and colorectal cancer, where Gankyrin interaction with MAGE-A4 reduces the oncogenic potential of tumour cells. In this study I aimed to investigate the heterogeneity of GCNIS in relation to disease stage and Sertoli cell development. We also aimed to determine the role of Gankyrin in TGCC cell survival and invasion. The co-expression of early germ cells proteins such as OCT4, LIN28 and PLAP was characterized in GCNIS cells during childhood and adulthood pre-invasive TGCC and in invasive disease characterized by the presence of a testicular tumour. These results show that LIN28 was expressed in 95% of OCT4 GCNIS cells, whereas PLAP expression in GCNIS cells increased as the disease progressed from childhood pre-invasive disease to invasive seminoma (32.3% v 76%; p < 0.05). In contrast there was a reduction in the proportion of MAGE-A4 expressing GCNIS cells with disease progression. The MAGE-A4 expressing population was also less proliferative than the MAGE-A4 negative GCNIS population. The methylation status of GCNIS cells was then investigated. EZH2 a methyltransferase previously reported to be important for TGCC development, was expressed in GCNIS cells at all stages of disease, however the histone 3 modification H3K27me3 (mediated by EZH2) was expressed in a significantly higher percentage of the proliferative OCT4+/MAGE-A4- GCNIS cells compared with the OCT4+/MAGEA4+ population (11.7% v 1.1%; p < 0.01) which could indicate a repressive role for H3K27me3 over MAGE-A4 expression. Next, it was determined whether an association between Sertoli cell maturation status and progression of TGCC could be observed. The maturation status of Sertoli cells was studied using proteins indicative of immature (desmin, cytokeratin, fibronectin and AMH) and mature (vimentin and androgen receptor) Sertoli cells. These studies demonstrated heterogeneity of Sertoli cells maturation in GCNIS-containing tubules. Desmin, fibronectin, AMH and vimentin expression did not show any association with TGCC progression. Cytokeratin was expressed in Sertoli cells of human fetal testis up to second trimester of fetal life, absent in tubules with active spermatogenesis but heterogeneously present in GCNIS, demonstrating that cytokeratin expression is indicative of the presence of GCNIS. Androgen receptor was weakly present in Sertoli cells from human fetal testis and pre-pubertal pre-invasive TGCC testis whereas in GCNIS of adult pre-invasive testis and invasive samples, androgen receptor was abundantly expressed in Sertoli cells of GCNIS-containing tubules. These combined results for cytokeratin and androgen receptor suggest that Sertoli cells from GCNIS-containing tubules, in pre-invasive and invasive TGCC patients are partially differentiated. Gankyrin expression was characterised in fetal germ cells, GCNIS cells and TGCC tissue. In fetal testis nuclear Gankyrin was absent in OCT4+/MAGE-A4- (gonocyte) population whereas it was present in a subpopulation of OCT4-/MAGE-A4+ (spermatogonia) germ cells. In GCNIS cells from TGCC patients nuclear Gankyrin was expressed in 87%, 63.3%, 91.5% and 79% in childhood pre-invasive, adult pre-invasive, seminoma and non-seminoma GCNIS cells respectively. Finally, in seminoma cells, Gankyrin was expressed in the cytoplasm indicating a change in localisation as the GCNIS cells become invasive. We used siRNA to knockdown Gankyrin in NT2 (a TGCC cell line) cells in-vitro and demonstrated a decrease in cell number, suggesting that Gankyrin might play a role in TGCC progression and invasiveness. Gankyrin down-regulation also resulted in an increase in p53 and p21 mRNA level. Given the role of P53 and p21 in cisplatin cytotoxic effect in TGCC we went on to investigate the role of Gankyrin in cisplatin resistance using NT2 cells. We demonstrate that Gankyrin mediated cisplatin resistance through the p53/p21 pathway, upregulating apoptosis rates through BAX and FAS, whilst there was no effect on cell proliferation, cell cycle or cell migration. In conclusion, we have shown that GCNIS cells are heterogeneous and their phenotype can determine their oncogenic potential. We also show that Sertoli cells from GCNIS-containing tubules undergo partial differentiation displaying markers of immature and mature Sertoli cells, with a heterogeneous association of cytokeratin with GCNIS presence. We also demonstrate that the oncogene Gankyrin has a role in NT2 cells survival and cisplatin resistance indicating that manipulation of Gankyrin may have a role in the treatment of TGCC.

Introdução: A incidência de câncer anal vem aumentando nas últimas décadas principalmente em determinados grupos de risco como pacientes HIV positivos e homens homossexuais e bissexuais. A neoplasia intraepitelial anal (NIA) representa a lesão precursora do câncer anal e tem demonstrado clara associação com o papilomavírus humano (HPV) de alto risco. Mulheres com neoplasia intraepitelial ou carcinoma invasor genital parecem ter um risco aumentado para câncer anal. O objetivo deste estudo é determinar a prevalência de neoplasia intraepitelial anal neste grupo de mulheres. Métodos: Estudo transversal onde 106 mulheres imunocompetentes com diagnóstico histológico de neoplasia intraepitelial ou câncer genital e 74 pacientes sem neoplasia genital foram submetidas à anuscopia de alta resolução com biópsia de áreas alteradas. Resultados: A prevalência geral de NIA na amostra foi 6,6%. No grupo de mulheres com neoplasia genital foi 10,4% (IC 95%, 5,6 a 17,3%) e 1,4% (IC 95%, 0,07 a 6,5%) nas mulheres sem esta condição (p=0,016). A razão de prevalência encontrada foi 7,68 (IC 95%, 1,01 a 58,21) e a razão de chances foi 8,45 (IC 95%, 1,07 a 66,97). Conclusão: A prevalência de NIA foi maior em mulheres com neoplasia intraepitelial ou câncer invasor genital. / Background: In the last decade the incidence of anal cancer is increasing especially in high risk groups as those infected with the human immunodeficiency virus (HIV) and men who have sex with men (MSM). Anal intraepithelial neoplasia (AIN) is believed to be a precursor of anal cancer. It appears to be related to high-risk human papillomavirus (HPV). Women with genital intraepithelial neoplasia or cancer have shown to be at increased risk for anal cancer. The aim of this study is to determine the prevalence of anal intraepithelial neoplasia in this group of women. Methods: In a cross sectional study, 106 imunocompetent women with histopathological diagnosis of genital intraepithelial neoplasia or cancer and 74 women without gynecologic neoplasia underwent to high-resolution anoscopy with biopsy of visible lesions. Results: The overall prevalence of AIN was 6.6%. In women with genital neoplasia it was 10.4% (95% CI, 5.6 to 17.3%) and 1.4% (95% CI, 0.07 to 6.5%) in women without genital neoplasia (p=0.016). The prevalence ratio was 7.68 (95% CI, 1.01 to 58.21) and the odds ratio 8.45 (IC 95%, 1.07 a 66.97). Conclusion: The prevalence of AIN was higher in women with genital intraepithelial neoplasia or cancer.

Introdução: A incidência de câncer anal vem aumentando nas últimas décadas principalmente em determinados grupos de risco como pacientes HIV positivos e homens homossexuais e bissexuais. A neoplasia intraepitelial anal (NIA) representa a lesão precursora do câncer anal e tem demonstrado clara associação com o papilomavírus humano (HPV) de alto risco. Mulheres com neoplasia intraepitelial ou carcinoma invasor genital parecem ter um risco aumentado para câncer anal. O objetivo deste estudo é determinar a prevalência de neoplasia intraepitelial anal neste grupo de mulheres. Métodos: Estudo transversal onde 106 mulheres imunocompetentes com diagnóstico histológico de neoplasia intraepitelial ou câncer genital e 74 pacientes sem neoplasia genital foram submetidas à anuscopia de alta resolução com biópsia de áreas alteradas. Resultados: A prevalência geral de NIA na amostra foi 6,6%. No grupo de mulheres com neoplasia genital foi 10,4% (IC 95%, 5,6 a 17,3%) e 1,4% (IC 95%, 0,07 a 6,5%) nas mulheres sem esta condição (p=0,016). A razão de prevalência encontrada foi 7,68 (IC 95%, 1,01 a 58,21) e a razão de chances foi 8,45 (IC 95%, 1,07 a 66,97). Conclusão: A prevalência de NIA foi maior em mulheres com neoplasia intraepitelial ou câncer invasor genital. / Background: In the last decade the incidence of anal cancer is increasing especially in high risk groups as those infected with the human immunodeficiency virus (HIV) and men who have sex with men (MSM). Anal intraepithelial neoplasia (AIN) is believed to be a precursor of anal cancer. It appears to be related to high-risk human papillomavirus (HPV). Women with genital intraepithelial neoplasia or cancer have shown to be at increased risk for anal cancer. The aim of this study is to determine the prevalence of anal intraepithelial neoplasia in this group of women. Methods: In a cross sectional study, 106 imunocompetent women with histopathological diagnosis of genital intraepithelial neoplasia or cancer and 74 women without gynecologic neoplasia underwent to high-resolution anoscopy with biopsy of visible lesions. Results: The overall prevalence of AIN was 6.6%. In women with genital neoplasia it was 10.4% (95% CI, 5.6 to 17.3%) and 1.4% (95% CI, 0.07 to 6.5%) in women without genital neoplasia (p=0.016). The prevalence ratio was 7.68 (95% CI, 1.01 to 58.21) and the odds ratio 8.45 (IC 95%, 1.07 a 66.97). Conclusion: The prevalence of AIN was higher in women with genital intraepithelial neoplasia or cancer.

Introdução: A incidência de câncer anal vem aumentando nas últimas décadas principalmente em determinados grupos de risco como pacientes HIV positivos e homens homossexuais e bissexuais. A neoplasia intraepitelial anal (NIA) representa a lesão precursora do câncer anal e tem demonstrado clara associação com o papilomavírus humano (HPV) de alto risco. Mulheres com neoplasia intraepitelial ou carcinoma invasor genital parecem ter um risco aumentado para câncer anal. O objetivo deste estudo é determinar a prevalência de neoplasia intraepitelial anal neste grupo de mulheres. Métodos: Estudo transversal onde 106 mulheres imunocompetentes com diagnóstico histológico de neoplasia intraepitelial ou câncer genital e 74 pacientes sem neoplasia genital foram submetidas à anuscopia de alta resolução com biópsia de áreas alteradas. Resultados: A prevalência geral de NIA na amostra foi 6,6%. No grupo de mulheres com neoplasia genital foi 10,4% (IC 95%, 5,6 a 17,3%) e 1,4% (IC 95%, 0,07 a 6,5%) nas mulheres sem esta condição (p=0,016). A razão de prevalência encontrada foi 7,68 (IC 95%, 1,01 a 58,21) e a razão de chances foi 8,45 (IC 95%, 1,07 a 66,97). Conclusão: A prevalência de NIA foi maior em mulheres com neoplasia intraepitelial ou câncer invasor genital. / Background: In the last decade the incidence of anal cancer is increasing especially in high risk groups as those infected with the human immunodeficiency virus (HIV) and men who have sex with men (MSM). Anal intraepithelial neoplasia (AIN) is believed to be a precursor of anal cancer. It appears to be related to high-risk human papillomavirus (HPV). Women with genital intraepithelial neoplasia or cancer have shown to be at increased risk for anal cancer. The aim of this study is to determine the prevalence of anal intraepithelial neoplasia in this group of women. Methods: In a cross sectional study, 106 imunocompetent women with histopathological diagnosis of genital intraepithelial neoplasia or cancer and 74 women without gynecologic neoplasia underwent to high-resolution anoscopy with biopsy of visible lesions. Results: The overall prevalence of AIN was 6.6%. In women with genital neoplasia it was 10.4% (95% CI, 5.6 to 17.3%) and 1.4% (95% CI, 0.07 to 6.5%) in women without genital neoplasia (p=0.016). The prevalence ratio was 7.68 (95% CI, 1.01 to 58.21) and the odds ratio 8.45 (IC 95%, 1.07 a 66.97). Conclusion: The prevalence of AIN was higher in women with genital intraepithelial neoplasia or cancer.

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No. of bitstreams: 2 Dissertação - Nayara Alves de Freitas Lemos - 2015.pdf: 1558373 bytes, checksum: f3d20ea70bfd08864a3c3eaae4b69cad (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-07-17 / Objective: To analyze the temporal evolution of DCIS in residents of Goiânia during the period 1994-2010. Methods: It used the database of the Population Based Cancer Registries of Goiania (RCBPGo), cases coded as carcinoma in situ of the breast in females, at (ONCOSIS) program in Goiânia, between 1994 and 2010. It was later made individual search of histopathological reports of DCIS. We sought to identify the temporal evolution of standardized and crude incidence of DCIS. The incidence rates, crude, as standard, set by the global population Doll, were calculated by age groups to 10 years from 30 years, and was estimated to MPMA using Poisson Regression to these age groups. They calculated the disease-free survival and overall survival at 60 and 120 months, using the Kaplan-Meier method. The data on DCIS deaths were obtained from the Mortality Data System (SIM), the medical record information and the electoral higher court (TSE). Results: In the initial database were recorded 376 cases of DCIS. In reviewing the reports, 114 cases were excluded because it is not dealt with DCIS. Of the 282 cases of DCIS in the period studied, there were four cases in 1994 and 21 in 2010. The crude rate of annual incidence of DCIS was 1.33/100,000 in 1994, and of 4.21/100,000 in 2010. The adjusted incidence for the world population Doll was 0.58/100,000 in 1994, and of 1.85/100,000 in 2010. The average annual percentual change (AAPC) of the crude incidence rate for the period was 11.93% per year (95 9-15% CI; P <0.01) and standardized incidence rate of 11.94% per year (95% CI 9-15; p <0.01). There were 17 cases of local recurrence, 16 invasive ductal carcinomas and only one case of in situ recurrence. Three cases evolved with distant metastases. The cumulative rate of local recurrence was 3,9% at 60 months and 10% to 120 months. Overall survival was 96,5% and 91,9% at 60 and 120 months, respectively. The cancer-specific survival was 99,5% at 60 months and 98,4% at 120 months. Abstract xvii Conclusions: The study showed that there are a large number of cases that need to be recoded by changing the initial bank. Thus, we suggest that the highest injury potential aggressiveness is described first, standardized reports and the training of collectors, so there are no unknown information to transcribe the DCIS for RCBP the chips. There was an increasting incidence of DCIS rate in Goiânia, possibly related to mammographic screening. Despite the small number of local recurrences when appeared they arose mostly with invasion. Still, it was confirmed in the studied group high overall survival rate after 10 years of treatment. / Objetivo: Analisar a evolução temporal do carcinoma ductal in situ em moradores de Goiânia durante o período de 1994 a 2010. Métodos: Trata-se de um estudo descritivo retrospectivo de série temporal dos casos de carcinoma ductal in situ, no sexo feminino, em Goiânia, registrados no banco de dados do Registro de Câncer de Base Populacional dessa cidade no período entre 1994 a 2010. Posteriormente, realizou-se busca individual dos laudos histopatológicos de carcinoma ductal in situ para identificar a evolução temporal do carcinoma ductal in situ. As taxas de incidências, tanto bruta, quanto padronizada, ajustada pela população padrão, foram calculadas por grupos etários a cada 10 anos, a partir de 30 anos, e calculou-se a mudança percentual da média anual utilizando-se a regressão de Poisson. Para a análise de sobrevida global foi realizada busca ativa das pacientes no Sistema de Informações em Mortalidade, nas informações do prontuário médico e no Tribunal Superior Eleitoral. Foram calculadas a sobrevida livre de doença e a sobrevida global em 60 e 120 meses, pelo método de Kaplan-Meier. Resultados: No banco de dados inicial foram registrados 376 casos de CDIS. Na revisão dos laudos, foram excluídos 114 casos, pois não faziam parte dos critérios de inclusão. Dos 262 casos em Goiânia no período estudado, houve quatro casos em 1994 e 21 em 2010. A taxa bruta de incidência anual de CDIS foi 1,33/100.000 em 1994, e de 4,21/100.000 em 2010. Já a incidência ajustada para a população padrão foi de 0,58/100.000 em 1994, e de 1,85/100.000 em 2010. A mudança percentual da média anual da taxa de incidência bruta para o período foi de 11,93% ao ano (95% IC 9-15; p<0,01) e da taxa de incidência padronizada de 11,94% ao ano (95% IC 9 - 15; p<0,01). Houve 1 7 casos de recidiva local, sendo 1 6 carcinomas ductal invasores e apenas um caso de recidiva in situ. Três casos evoluíram com metástases à distância. A taxa cumulativa de recidiva local foi de 3,9% aos 60 meses e de 10% aos 120 meses. A sobrevida global Resumo xv foi de 96,5% e de 91,9% aos 60 e 120 meses, respectivamente. A sobrevida câncer-específica foi de 99,5% aos 60 meses e de 98,4% aos 120 meses. Conclusões: o trabalho mostrou um grande número de casos que precisam ser recodificados, alterando o banco inicial. A sugestão é que os laudos histopatológicos descrevam primeiramente a lesão de mais alto potencial de agressividade. É necessária uma padronização dos laudos, e a partir daí, o treinamento dos coletadores, para que não haja informações desconhecidas ao transcrever o CDIS para as fichas do RCBPGo. Foi constatado o aumento da taxa de incidência do CDIS na cidade de Goiânia, possivelmente relacionado à melhora do rastreamento mamográfico. E apesar do pequeno número de recidivas locais, quando apareciam, surgiam na sua grande maioria com invasão. Ainda assim, confirmou-se no grupo estudado alta taxa de sobrevida global após 10 anos do tratamento.

Orientadores: Sophie Françoise Mauricette Derchain, Luiz Carlos Zeferino / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-03T16:22:48Z (GMT). No. of bitstreams: 1 Torres_JoseCarlosCampos_D.pdf: 2435784 bytes, checksum: 57d4b3f48b5d1b95c38559b857c45c5f (MD5) Previous issue date: 2003 / Doutorado

Orientador: Liliana Aparecida Lucci de Angelo Andrade / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-07-19T13:41:15Z (GMT). No. of bitstreams: 1 Engelman_DianaEliciSader_M.pdf: 6002097 bytes, checksum: 33ceda50142d22ad0fcec1bfc9d0c09c (MD5) Previous issue date: 1994 / Resumo: Recentemente tem sido observado um aumento da freqüência de diagnósticos de neoplasia intraepitelial vulvar grau III, principalmente em mulheres jovens. Estudos sobre o câncer genital apontam evidências de que a infecção pelo vírus do papiloma humano é um fator de risco importante para a transformação neoplásica epitelial. No período de 1983 a 1992 foram levantados 30 casos de l1eoplasia intraepitelial vulvar grau III e seis de carcinoma escamoso estádio Ia. A pesquisa do antígeno comum do capsídeo viral foi realizada através da imulloperoxidase. O estudo do tipo do vírus nas lesões foi realizada pela técnica de hibridização "in situ", com utilização de sondas biotiniladas dos HPV 6/11, 16/18 e 31/35/51. A idade média das pacientes com neoplàsia intraepitelial vulvar foi de 47,5 anos e das com carcinoma estádio Ia, 61,1 anos. Histologicamente foram observadas .atipias coilocitóticas em 83,3% dos casos. O exame da imunoperoxidase foi positivo em 36,1% dos casos, com idade,' média de 39,1 anos. A detecção do vírus pela hibridização "in situ" ocorreu em 41,7% dos casos, com idade média de 38,3 anos. Todos os casos positivos apresentaram o tipo 16/18, porém em quatro casos houve associação de diferentes tipos virais na mesma lesão. A positividade dos métodos não estava associada à presença ou ausência de atipias coilocitóticas. Quando se considerou a positividade para um ou outro método, a detecção do vírus ocorreu em 63,9% dos casos. A alto índice de detecção relaciona o papilomavírus humano com estas lesões, em especial o tipo 16/18, principalmente em grupos etários mais jovens. A ampla faixa etária de distribuição dos casos de neoplasia intraepitelial vulvar grau lU, sugere que o seu tempo de evolução é longo, demonstrando a importância do exame da vulva no exame ginecológico de rotina para melhor detecção e tratamento precoce das lesões / Abstract: Recently, an increase has been observed in the frequency of diagnosis of vulvar intraepithelial neoplasia grade III, special1y in young women. Studies in gynecologic cancer suggest that hwnan papillomavirus infections play an important role in the epithelial neoplastic transfOlmation. Between 1983 and 1992, 30 cases of vulvar intraepithelial neoplasia grade III and six cases of vulvar carcinoma stage Ia were analysed. The research for the common antigen of the viral capsid was done by the immunoperoxidase technique. The virus types were identified by the "in situ" hybridization technique, with biotinylated probes for HPV 6/11, 16/18 and 31/35/51. The mean age ofthe patients with vulvat. int:raepithelial neoplasia was 47,5 years an the ones with vulvar carcinoma stage Ia, 61, I years. Histologically, koilocytotic atypia was observed in 83,3% of the cases. The .immunoperoxidase teclmique was positive in 36,1% of the cases, with the mean age of 39, 1 years. The detection of the virus by "in situ" hybridization occurred in 41,7% of the cases, with mean age of 38,3 years. AlI the positive cases had the virus type 16/18, but in four of them there was association with other virus types in the same lesion. The positivity of these techniques was not related to the presence or absence of koiIocytotic atypia. When the positivity for one or the other method was considered, virus detection occulTed in 63,9% of the cases. This high rate of detection shows a relationship of the human papillomavirus with these vulvar Iesions, specially with type 16/18 and in younger age groups. The wide age range distribution of the vulvar intraepithelial neopIasia grade lU patients, suggests that its evolution is long, showing the importance of the vulvar region in routine gynecologic examination, to allow early detection and treatment of these lesions / Mestrado / Anatomia Patologica / Mestre em Ciências Médicas

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18128号 / 医博第3848号 / 新制||医||1001(附属図書館) / 30986 / 京都大学大学院医学研究科医学専攻 / (主査)教授 福山 秀直, 教授 戸井 雅和, 教授 平岡 眞寛 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DGAM

Changes in the microenvironment have been implicated in the transition of pre-invasive ductal carcinoma in-situ (DCIS) to invasive breast cancer. Normal myoepithelial cells have a tumour suppressor phenotype but they are altered in DCIS and ultimately lost with transition to invasive cancer. A consistent change in DCIS is up-regulation of the integrin αvβ6 in myoepithelial cells. Preliminary observations identified a correlation between myopeithelial αvβ6 and an increased peri-ductal inflammatory infiltrate. The hypothesis of this study is that the altered myoepithelial phenotype influences the peri-ductal inflammatory environment, which in turn mediates a pro-apoptotic effect on myoepithelial cells contributing to their loss. To investigate this, the inflammatory infiltrate was characterised in a series of DCIS tissue in relation to αvβ6 status. This demonstrated significantly higher levels of CD4+ve and FOXP3+ve T cells around αvβ6+ve DCIS ducts compared to αvβ6-ve ducts (P=<0.01), suggesting an increase in Treg cells. In-vivo, Matrigel plugs containing injected into the flanks of female C57/Blk6 normal mice generated influx of higher levels of CD4+ve cells (p=0.005) and FOXP3+ T cells (p=0.007) in the presence of αvβ6+ve myoepithelial cells compared to αvβ6-ve cells, supporting the findings in human tissue samples. Since Treg cells produce TRAIL that can induce apoptosis, we investigated the influence of αvβ6 on myoepithelial cells on the levels of TRAIL in T cells and the hypothesis that αvβ6-positive myoepithelial ells may be more susceptible to TRAIL-induced apoptosis, leading to loss of the myoepithelial barrier. Firstly, levels of TRAIL in Jurkat and primary T cell populations co-cultured with β4 (ii) or β6 myoepithelial cells were measured. This demonstrated a higher level of TRAIL in primary T cells co-cultured β6 myoepithelial cells compared to those co-cultured with β4 myoepithelial cells. β6+ve and β6-ve myoepithelial cells were exposed to TRAIL, and this demonstrated that TRAIL enhanced apoptosis, measured by cleaved PARP, in β6+ve cells. Furthermore, these cells showed loss of the anti-apoptotic protein Galectin-7, and knockdown of Galectin-7 in normal β6-ve myoepithelial cells rendered them more susceptible to TRAIL-induced apoptosis. In DCIS tissues, an inverse relationship between αvβ6 and Galectin-7 in myoepithelial cells was demonstrated, and Cytokine Array analysis showed that αvβ6+ve myoepithelial cells express higher levels of IL-16, which has a role in Treg cell recruitment. Taken together these results suggest that expression of αvβ6 by myoepithelial cells in DCIS generates a tumour-promoter peri-ductal inflammatory infiltrate through altered cytokine release, is associated with reduced galectin-7 expression and enhances myoepithelial cell apoptosis in response to TRAIL. This provides a potential mechanism by which myoepithelial cells may be lost during evolution of DCIS and so contribute to progression to invasive disease.

Background: A maioria dos métodos de rastreamento do câncer de colo de útero depende da colposcopia para a confirmação do diagnóstico. A colposcopia sofre com a falta de disponibilidade, a necessidade de longos deslocamentos por parte das pacientes e longas filas de espera. Objetivo: Avaliar o desempenho da Fotografia Cervical Digital como um método alternativo à colposcopia. Método: Foram realizadas colposcopies e Fotografias Digitais Cervicais em 228 pacientes. As Fotografias Digitais Cervicais foram avaliadas através da internet por três colposcopistas experientes. A concordância entre os métodos foi calculada através de Kappa e as porcentagens de concordância. Sensibilidade, especificidade, valor preditivo positivo, valor preditivo negativo e acuracia diagnóstica foram calculados para a colposcopia e a Fotografia Cervical Digital. A histologia foi utilizada como padrão-ouro. Resultados: A Fotografia Cervical Digital e a colposcopia concordaram em 89,9% dos casos (K=0,588). A Fotografia Cervical Digital e a colposcopia apresentaram desempenhos comparáveis, a sensibilidade foi de 52,5% e 35,0%, a especificidade de 91,86% e 91,28%, o valor preditivo positivo de 60,0% e 48,3%, valor preditivo negativo de 89,3% e 85,8%, e a acurácia diagnóstica de 84,4% e 80,7%, respectivamente. Conclusão: A Fotografia Cervical Digital é um método alternativo promissor à colposcopia. / Background: Most cervical cancer screening methods relies on colposcopy to confirm the diagnosis. Colposcopy suffers from the lack of availability, long patients’ displacement and waiting times. Objective: Evaluate the performance of the Cervical Digital Photography as an alternative method to colposcopy. Methods: Colposcopy and Cervical Digital Photography were performed in 228 women. The Cervical Digital Photographs were evaluated through internet by 3 colposcopy experts. The agreement between methods was calculated with kappa and percentages of agreement. Sensitivity, specificity, positive predictive value, negative predictive value and diagnostic accuracy were calculated for colposcopy and Cervical Digital Photography. Histology was the gold standard. Results: Cervical Digital Photography and colposcopy agreed in 89.9% of the cases (K=0.588). Cervical Digital Photography and colposcopy had comparable performances, sensitivity was 52.5% and 35.0%, specificity was 91.86% and 91.28%, positive predictive value was 60.0% and 48.3%, negative predictive value was 89.3% and 85.8%, and diagnostic accuracy was 84.4% and 80.7%, respectively. Conclusion: Cervical Digital Photography is a promising alternative method to colposcopy.