Es wurden 177 Patientinnen mit zwischen 1999 und 2005 an der Universitätsfrauenklinik Leipzig operativ therapiertem Mammakarzinom und vorangegangener Stanzbiopsie, für die vollständige Befunde vorlagen und bei denen keine neoadjuvante Chemotherapie stattfand, hinsichtlich der prognostischen und prädiktiven Faktoren und deren Vergleich zwischen Stanzbiopsie und dem endgültigen Tumorexzidat untersucht. Unsere Daten zeigten, dass die Stanzbiopsie in der Einschätzung des Differenzierungsgrades mit einer Konkordanz von 62,9% und der Lymphgefäßinvasion mit einer Konkordanz in 69,8% keine hohe Genauigkeit besitzt. Bezüglich des histologischen Typs mit einer Übereinstimmung von 77%, der Östrogen- und Progesteronrezeptorbestimmung mit Konkordanzen von 87% und 83% sowie hinsichtlich des Her-2/neu-Status mit einer Konkordanz von 79% fand sich eine moderate bis gute Übereinstimmung mit dem Exzidat, wobei zu diskutieren ist, ob bei initial an der Stanzbiopsie negativem Östrogen- und/oder Progesteronrezeptorstatus oder auch bei positivem Progesteronrezeptor- und negativem Östrogenrezeptornachweis eine erneute immunhistochemische Hormonrezeptoruntersuchung am Exzidat erfolgen sollte sowie ob bei einer Konkordanzrate des Her-2/neu von weniger 95% immer eine zweite Bestimmung am Operationspräparat als Basis einer definitiven Therapieplanung durchgeführt werden muß. In 8,5% wurde an der Biopsie keine Malignität festgestellt. Der Vergleich des Malignitätsgrades mit der Tumorkategorie als auch mit dem Lymphknotenstatus zeigte keine signifikante Korrelation. Eine fortgeschrittene Tumorkategorie war mit dem Vorhandensein von Lymphknotenmetastasen korreliert.

XII tarptautinėje St Gallen krūties vėžio konferencijoje (2011) Ekspertų komisijos priimta nauja pacientų klasifikacija sisteminei terapijai atlikti, paremta biologiniais krūties vėžio subtipais, kurie apibrėžiami imunohistocheminiu tyrimu (IHC). Tačiau ši nauja navikų klasifikacija iš esmės pagrįsta pusiau kiekybiniu biožymenų raiškos vertinimu, todėl išlieka aktuali ribinių verčių nustatymo problematika. Esminiai pokyčiai IHC tyrimų srityje galimi atsiradus skaitmeninio vaizdinimo technologijoms, leidžiančiomis IHC tyrimų rezultatus analizuoti kiekybiniais parametrais. Darbe naudojant skaitmeninį vaizdo analizės metodą atliktas išsamus biologinių žymenų tyrimas leido palyginti svarbių, tačiau nepakankamai ištirtų (p53, AR, p16, BCL2, SATB1, HIF1) IHC žymenų informatyvumą su esamų prognozinių žymenų (ER, PR, HER2, Ki67) rodikliais. Ištirtas platus genetinių ir epigenetinių krūties vėžio žymenų spektras. Pirmą kartą 10 IHC žymenų rinkinio, įvertinto skaitmeninės analizės būdu, duomenys panaudoti faktorinės analizės metodu nustatyti jų variacijų vidinius veiksnius, atskleidžiančius biologinius dėsningumus ir IHC žymenų bei jų derinių informatyvumą. Šios analizės rezultatai leido naujai įvertinti publikuotų krūties vėžio IHC žymenų bei jų derinių (p16, SATB1, HIF1, Ki67/BCL2 ir kt.) informatyvumą. / The 12th International St Gallen conference on breast cancer (2011) proposed patient categorization for systemic therapy, based on intrinsic breast cancer subtypes, defined by imunohistochemistry (IHC) test results. Since this classification is based on semi-quantitative expression of IHC biomarker expression, an issue of defining and applying cutoff values remains. Essential improvement in the IHC testing has become possible with digital image analysis tools enabling quantitative evaluation of the IHC data. This study explores data obtained by digital image analysis methods applied to evaluate a comprehensive biomarker dataset (p53, AR, p16, BCL2, SATB1, HIF1) along with well established (ER, PR, HER2, Ki67) biomarkers. Also, an extensive set of genetic and epigenetic biomarkers has been tested. For the first time, the dataset of 10 IHC biomarkers, evaluated by digital analysis was explored by the means of factor analysis to establish the intrinsic factors of biological variation and informative value of IHC biomarkers and their combinatiions. The results also provided insights into the significance and combinatorial effects of the established and relatively new biomarkers (p16, SATB1, HIF1, Ki67/BCL2, etc.).

Introdução: O câncer de mama é a neoplasia mais freqüente e também a principal causa de morte por câncer entre as mulheres. O gene TP53 é polimórfico no códon 72 da proteína que ele codifica, podendo conter arginina (CGC) ou prolina (CCC) nesta posição. Este polimorfismo pode estar envolvido na suscetibilidade e predisposição ao câncer e apresenta uma distribuição étnica e geográfica bastante variável. O genótipo homozigoto para arginina parece ser um fator de risco e prognóstico significativo para o câncer de mama. Métodos: Extraído DNA a partir do sangue periférico de 76 pacientes consecutivas com carcinoma ductal invasor (CDI), tratadas no Serviço de Mastologia do HCPA, em qualquer estágio da doença. 80 amostras de DNA do grupo controle de doadores saudáveis do Banco de Sangue do HCPA foram incluídas de forma aleatória. Foram coletados dados demográficos e dados das características clínicas e histopatológicas e realizada a amplificação do éxon 4 do gene TP53 através da PCR, seguida da identificação do polimorfismo R72P do éxon 4 pela digestão do produto de PCR com a enzima de restrição BstUI, a qual reconhece o sítio de clivagem CG↓CG.Os objetivos foram determinar as freqüências alélicas e genotípicas do polimorfismo R72P nas pacientes com carcinoma de mama ductal invasor, comparando-as com as freqüências no grupo controle e relacionar a presença deste polimorfismo com características clínicopatológicas. Resultados: A distribuição dos genótipos no códon 72 do gene TP53, tanto em pacientes como em controles, encontra-se em equilíbrio de Hardy- Weinberg. A freqüência encontrada para o polimorfismo R72P foi similar entre as pacientes com CDI e os controles, não sendo encontrada diferença significativa na freqüência do genótipo (P = 0,707) e na freqüência alélica (P = 0,469). Conclusões: O polimorfismo R72P no gene TP53 não se associou a maior risco de desenvolvimento de CDI na população estudada. Este achado pode estar relacionado à grande variação inter-racial e étnica em nossa população, decorrente de freqüentes miscigenações e exposição a diferentes fatores ambientais, importantes na evolução do carcinoma de mama. Não houve associação com características clínico-patológicas. / Introduction: Breast cancer is the most frequent neoplasia as well as the main cause of death from cancer among women. The TP53 gene is polymorphic in codon 72 of the protein it encodes, and may contain either arginine (CGC) or proline (CCC) in that position. Such polymorphism may be involved in the susceptibility and predisposition for cancer presenting with a widely variable ethnic and geographic distribution. The arginine homozygous genotype seems to be a significant risk factor and prognostic for breast cancer. Methods: DNA was extracted from peripheral blood of 76 patients suffering from invasive ductal carcinoma (IDC) treated at the HCPA Mastology Service at any stage of the disease. Eigthy healthy controls from the Blood Bank Donors of HCPA were included randomly. Demographic data were collected as well as data from the clinic and histopathological characteristics. Amplification of exon 4 from the TP53 gene was performed through PCR, followed by digestion of the PCR product with the restriction enzyme BstUI, which recognizes the CG↓CG cleavage site, for identification of the R72P polymorphism of exon 4. The objectives were to determine the allele and genotype frequencies of R72P polymorphism in patients suffering from invasive ductal breast carcinoma, comparing with the control group and to corelate the presence of that polymorphism with clinical pathological characteristics. Results: The distribution of genotypes in codon 72 of gene TP53, both for patients and controls was in Hardy-Weinberg equilibrium. The frequency found for R72P polymorphism was similar between IDC patients and controls, with no significant difference being found in the genotype frequency (P = 0.707) and allele frequency (P = 0.469). Conclusions: R72P polymorphism in the TP53 gene was not associated to an increased risk of developing IDC in the population studied. This finding may be related to the great interracial and ethnic variation in our population deriving from frequent miscigenations and exposure to different environmental factors that are important in the evolution of breast cancer. There was also no association with clinical pathological characteristics.

Made available in DSpace on 2014-06-11T19:32:50Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-08-31Bitstream added on 2014-06-13T21:04:58Z : No. of bitstreams: 1 delgallo_wd_dr_botfm.pdf: 223230 bytes, checksum: 254dbff0ba8e45f49db67178b8a1ae49 (MD5) / Estudos de expressão gênica têm identificado vários grupos moleculares de carcinoma de mama, com diferentes comportamentos clínico e biológico. A correlação entre “cDNA microarray” e imunoistoquímica(IQ) com marcadores para citoceratinas, Her2/neu, receptor de estrógeno(RE) e de células basais mioepiteliais (1A4, S-100 e p63), identificaram cinco grupos: (1) luminal A (RE+; Her2/neu-), (2) luminal B (RE+; Her2/neu+), (3) superexpressão de Her2/neu (RE-; Her2/neu+), (4) tipo basal (RE-; Her2/neu-; Ck 5/6 +) e (5) nenhum destes (“null”). Os de tipo luminal expressam citoceratinas de padrão luminal (Ck8/18) e os de tipo basal expressam citoceratinas 5/6 e 14 ou marcadores de células basais mioepiteliais. Avaliamos a expressão de Ck5/6, Ck8/18 e 1A4 em material de citoinclusão, comparando-a ao espécime cirúrgico. Material e Métodos: Foram selecionados 62 casos, seqüenciais, de carcinoma de mama diagnosticados por PAAF, com citoinclusão e espécime cirúrgico. Cortes de citoinclusão e do espécime cirúrgico foram imunocorados para Ck 5/6, Ck 8/18 e 1A4. Resultados e Conclusão: Os valores, em porcentagem, de sensibilidade, especificidade, valor preditivo positivo(VPP), valor preditivo negativo(VPN) e acurácia foram, respectivamente: Ck5/6 (77, 100, 100, 92 e 94); Ck8/18 ( 98, 66, 96, 80 e 95) e 1A4 ( 92, 96, 85, 98 e 95). Portanto, a identificação de Ck5/6, Ck8/18 e 1A4 por IQ em material de citoinclusão é método confiável, com resultados muito próximos aos obtidos no espécime cirúrgico e pode contribuir para a classificação dos carcinomas mamários de expressão luminal e basal, fornecendo informações importantes que possam orientar na escolha do tratamento, bem como na avaliação de fatores prognósticos e preditivos. A importância da obtenção de dados morfológicos e imunoistoquímicos sobre os carcinomas mamários através do material... (Rewsumo completo, clicar acesso eletrônico abaixo) / Genetic expression studies have identified many molecular groups of breast carcinoma, with different clinical and biological behavior. The correlation between cDNA microarray and immunohistochemistry (IHC) with markers for cytokeratin, Her2/neu, estrogen receptor (ER) and of basal myoepithelial cells (1A4, S-100 e p63), identified five groups: (1) luminal A (ER+; Her2/neu-), (2) luminal B (ER+; Her2/neu+), (3) overexpression of Her2/neu (ER- ; Her2/neu+), (4) basal-like (ER- ; Her2/neu-; Ck 5/6 +) and (5) none of them (null). The luminal-like express cytokeratines of luminal pattern (Ck8/18) and the basal-like express cytokeratines 5/6 and 14 or markers of myoepithelial basal cells. We have evaluated the expression of Ck5/6, Ck8/18 and 1A4 in cell block comparing it to the surgical specimen. Material and Methods: 43 62 cases have been selected, sequencial, of breast carcinoma diagnosed through fine needle aspiration (FNA), with cell block and surgical specimen. Cuts of cell block and from the surgical specimen were immunostained for Ck 5/6, Ck 8/18 and 1A4. The value, in percentage, of sensibility, specificity, positive predictive value, negative predictive value, and accuracy were respectively: Ck5/6 (77, 100, 100, 92 e 94); Ck8/18 (98, 66, 96, 80 e 95) e 1A4 ( 92, 96, 85, 98 e 95). Therefore, the identification of CK5/6, 8/18 and 1A4 for IHC in cell block is a reliable method, with results very close to the ones obtained in the surgical specimen, and it can contribute to the sub classification of the breast carcinomas of luminal and basal expression, providing important information, which can orientate the treatment... (Complete abstract click electronic access below)

Introdução: O câncer de mama é a neoplasia mais freqüente e também a principal causa de morte por câncer entre as mulheres. O gene TP53 é polimórfico no códon 72 da proteína que ele codifica, podendo conter arginina (CGC) ou prolina (CCC) nesta posição. Este polimorfismo pode estar envolvido na suscetibilidade e predisposição ao câncer e apresenta uma distribuição étnica e geográfica bastante variável. O genótipo homozigoto para arginina parece ser um fator de risco e prognóstico significativo para o câncer de mama. Métodos: Extraído DNA a partir do sangue periférico de 76 pacientes consecutivas com carcinoma ductal invasor (CDI), tratadas no Serviço de Mastologia do HCPA, em qualquer estágio da doença. 80 amostras de DNA do grupo controle de doadores saudáveis do Banco de Sangue do HCPA foram incluídas de forma aleatória. Foram coletados dados demográficos e dados das características clínicas e histopatológicas e realizada a amplificação do éxon 4 do gene TP53 através da PCR, seguida da identificação do polimorfismo R72P do éxon 4 pela digestão do produto de PCR com a enzima de restrição BstUI, a qual reconhece o sítio de clivagem CG↓CG.Os objetivos foram determinar as freqüências alélicas e genotípicas do polimorfismo R72P nas pacientes com carcinoma de mama ductal invasor, comparando-as com as freqüências no grupo controle e relacionar a presença deste polimorfismo com características clínicopatológicas. Resultados: A distribuição dos genótipos no códon 72 do gene TP53, tanto em pacientes como em controles, encontra-se em equilíbrio de Hardy- Weinberg. A freqüência encontrada para o polimorfismo R72P foi similar entre as pacientes com CDI e os controles, não sendo encontrada diferença significativa na freqüência do genótipo (P = 0,707) e na freqüência alélica (P = 0,469). Conclusões: O polimorfismo R72P no gene TP53 não se associou a maior risco de desenvolvimento de CDI na população estudada. Este achado pode estar relacionado à grande variação inter-racial e étnica em nossa população, decorrente de freqüentes miscigenações e exposição a diferentes fatores ambientais, importantes na evolução do carcinoma de mama. Não houve associação com características clínico-patológicas. / Introduction: Breast cancer is the most frequent neoplasia as well as the main cause of death from cancer among women. The TP53 gene is polymorphic in codon 72 of the protein it encodes, and may contain either arginine (CGC) or proline (CCC) in that position. Such polymorphism may be involved in the susceptibility and predisposition for cancer presenting with a widely variable ethnic and geographic distribution. The arginine homozygous genotype seems to be a significant risk factor and prognostic for breast cancer. Methods: DNA was extracted from peripheral blood of 76 patients suffering from invasive ductal carcinoma (IDC) treated at the HCPA Mastology Service at any stage of the disease. Eigthy healthy controls from the Blood Bank Donors of HCPA were included randomly. Demographic data were collected as well as data from the clinic and histopathological characteristics. Amplification of exon 4 from the TP53 gene was performed through PCR, followed by digestion of the PCR product with the restriction enzyme BstUI, which recognizes the CG↓CG cleavage site, for identification of the R72P polymorphism of exon 4. The objectives were to determine the allele and genotype frequencies of R72P polymorphism in patients suffering from invasive ductal breast carcinoma, comparing with the control group and to corelate the presence of that polymorphism with clinical pathological characteristics. Results: The distribution of genotypes in codon 72 of gene TP53, both for patients and controls was in Hardy-Weinberg equilibrium. The frequency found for R72P polymorphism was similar between IDC patients and controls, with no significant difference being found in the genotype frequency (P = 0.707) and allele frequency (P = 0.469). Conclusions: R72P polymorphism in the TP53 gene was not associated to an increased risk of developing IDC in the population studied. This finding may be related to the great interracial and ethnic variation in our population deriving from frequent miscigenations and exposure to different environmental factors that are important in the evolution of breast cancer. There was also no association with clinical pathological characteristics.

Resumo: Foram selecionados 62 casos de carcinoma de mama, diagnosticados por punção aspirativa por agulha fina (PAAF), com confirmação pelo espécime cirúrgico. Citoinclusão e espécime cirúrgico foram submetidos à reação imunoistoquímica (IQ) para receptor de estrógeno (RE), de progesterona (RP) e proteína Her-2/neu. Consideraram-se positivos para RE e RP, casos com 1% ou mais de células coradas. Positividade para Her-2/neu foi avaliada numa escala de 0 a 3+. O índice de concordância entre citoinclusão e espécime cirúrgico variou de 90 a 94%. Sensibilidade, especificidade, valores preditivo positivo (VPP) e negativo (VPN) e acurácia da citoinclusão na pesquisa para os RE e RP e proteína Her-2/neu (3+) foram respectivamente: RE (92,7%; 85,7%; 92,7%; 85,7% e 90,3%), RP (92,7%; 94,7%; 97,4%; 87,0% e 93,5%) e Her-2/neu (70,0%; 100,0%; 100,0%; 94,5% e 95,2%). A citoinclusão apresenta excelente correlação com o espécime cirúrgico na pesquisa por IQ dos receptores hormonais e da proteína Her-2/neu. / Abstract: It was selected 62 cases of breast carcinoma, diagnosed by fine needle aspiration (FNA), with confirmation by surgical specimen. Cell-block and surgical specimen were submitted to immunohistochemistry (IHC) reaction, for estrogen receptor (ER), progesterone receptor (PR) and Her-2/neu protein. Cases with 1% or more cells stained were considered positive for ER and PR. Her-2/neu Positivity was evaluated by a score from 0 to 3+. The concordance index between cell-block and surgical specimen had a variation of 90 to 94%. Sensibility, specificity, predictive positive and negative values and accuracy for ER, PR and Her-2/neu protein, was respectively: ER (92,7%; 85,7%; 92,7%; 85,7% and 90,3%), PR (92,7%; 94,7%; 97,4%; 87,0% and 93,5%) and Her- 2/neu (70,0%; 100,0%; 100,0%; 94,5% and 95,2%). Cell-block shows excellent correlation with surgical specimen in assessment of hormonal receptors and Her-2/neu protein, by IHC. / Orientador: Rosa Marlene Viero / Coorientador: Cléverson Teixeira Soares / Banca: Rosa Marlene Viero / Banca: Gilberto Uemura / Banca: Ulisses Frederigue Junior / Mestre

Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-28Bitstream added on 2014-06-13T19:36:18Z : No. of bitstreams: 1 bueno_sp_me_botfm.pdf: 539455 bytes, checksum: 0cf36502a3658cc125b5e07c71274053 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Foram selecionados 62 casos de carcinoma de mama, diagnosticados por punção aspirativa por agulha fina (PAAF), com confirmação pelo espécime cirúrgico. Citoinclusão e espécime cirúrgico foram submetidos à reação imunoistoquímica (IQ) para receptor de estrógeno (RE), de progesterona (RP) e proteína Her-2/neu. Consideraram-se positivos para RE e RP, casos com 1% ou mais de células coradas. Positividade para Her-2/neu foi avaliada numa escala de 0 a 3+. O índice de concordância entre citoinclusão e espécime cirúrgico variou de 90 a 94%. Sensibilidade, especificidade, valores preditivo positivo (VPP) e negativo (VPN) e acurácia da citoinclusão na pesquisa para os RE e RP e proteína Her-2/neu (3+) foram respectivamente: RE (92,7%; 85,7%; 92,7%; 85,7% e 90,3%), RP (92,7%; 94,7%; 97,4%; 87,0% e 93,5%) e Her-2/neu (70,0%; 100,0%; 100,0%; 94,5% e 95,2%). A citoinclusão apresenta excelente correlação com o espécime cirúrgico na pesquisa por IQ dos receptores hormonais e da proteína Her-2/neu. / It was selected 62 cases of breast carcinoma, diagnosed by fine needle aspiration (FNA), with confirmation by surgical specimen. Cell-block and surgical specimen were submitted to immunohistochemistry (IHC) reaction, for estrogen receptor (ER), progesterone receptor (PR) and Her-2/neu protein. Cases with 1% or more cells stained were considered positive for ER and PR. Her-2/neu Positivity was evaluated by a score from 0 to 3+. The concordance index between cell-block and surgical specimen had a variation of 90 to 94%. Sensibility, specificity, predictive positive and negative values and accuracy for ER, PR and Her-2/neu protein, was respectively: ER (92,7%; 85,7%; 92,7%; 85,7% and 90,3%), PR (92,7%; 94,7%; 97,4%; 87,0% and 93,5%) and Her- 2/neu (70,0%; 100,0%; 100,0%; 94,5% and 95,2%). Cell-block shows excellent correlation with surgical specimen in assessment of hormonal receptors and Her-2/neu protein, by IHC.

Introdução: O câncer de mama é a neoplasia mais freqüente e também a principal causa de morte por câncer entre as mulheres. O gene TP53 é polimórfico no códon 72 da proteína que ele codifica, podendo conter arginina (CGC) ou prolina (CCC) nesta posição. Este polimorfismo pode estar envolvido na suscetibilidade e predisposição ao câncer e apresenta uma distribuição étnica e geográfica bastante variável. O genótipo homozigoto para arginina parece ser um fator de risco e prognóstico significativo para o câncer de mama. Métodos: Extraído DNA a partir do sangue periférico de 76 pacientes consecutivas com carcinoma ductal invasor (CDI), tratadas no Serviço de Mastologia do HCPA, em qualquer estágio da doença. 80 amostras de DNA do grupo controle de doadores saudáveis do Banco de Sangue do HCPA foram incluídas de forma aleatória. Foram coletados dados demográficos e dados das características clínicas e histopatológicas e realizada a amplificação do éxon 4 do gene TP53 através da PCR, seguida da identificação do polimorfismo R72P do éxon 4 pela digestão do produto de PCR com a enzima de restrição BstUI, a qual reconhece o sítio de clivagem CG↓CG.Os objetivos foram determinar as freqüências alélicas e genotípicas do polimorfismo R72P nas pacientes com carcinoma de mama ductal invasor, comparando-as com as freqüências no grupo controle e relacionar a presença deste polimorfismo com características clínicopatológicas. Resultados: A distribuição dos genótipos no códon 72 do gene TP53, tanto em pacientes como em controles, encontra-se em equilíbrio de Hardy- Weinberg. A freqüência encontrada para o polimorfismo R72P foi similar entre as pacientes com CDI e os controles, não sendo encontrada diferença significativa na freqüência do genótipo (P = 0,707) e na freqüência alélica (P = 0,469). Conclusões: O polimorfismo R72P no gene TP53 não se associou a maior risco de desenvolvimento de CDI na população estudada. Este achado pode estar relacionado à grande variação inter-racial e étnica em nossa população, decorrente de freqüentes miscigenações e exposição a diferentes fatores ambientais, importantes na evolução do carcinoma de mama. Não houve associação com características clínico-patológicas. / Introduction: Breast cancer is the most frequent neoplasia as well as the main cause of death from cancer among women. The TP53 gene is polymorphic in codon 72 of the protein it encodes, and may contain either arginine (CGC) or proline (CCC) in that position. Such polymorphism may be involved in the susceptibility and predisposition for cancer presenting with a widely variable ethnic and geographic distribution. The arginine homozygous genotype seems to be a significant risk factor and prognostic for breast cancer. Methods: DNA was extracted from peripheral blood of 76 patients suffering from invasive ductal carcinoma (IDC) treated at the HCPA Mastology Service at any stage of the disease. Eigthy healthy controls from the Blood Bank Donors of HCPA were included randomly. Demographic data were collected as well as data from the clinic and histopathological characteristics. Amplification of exon 4 from the TP53 gene was performed through PCR, followed by digestion of the PCR product with the restriction enzyme BstUI, which recognizes the CG↓CG cleavage site, for identification of the R72P polymorphism of exon 4. The objectives were to determine the allele and genotype frequencies of R72P polymorphism in patients suffering from invasive ductal breast carcinoma, comparing with the control group and to corelate the presence of that polymorphism with clinical pathological characteristics. Results: The distribution of genotypes in codon 72 of gene TP53, both for patients and controls was in Hardy-Weinberg equilibrium. The frequency found for R72P polymorphism was similar between IDC patients and controls, with no significant difference being found in the genotype frequency (P = 0.707) and allele frequency (P = 0.469). Conclusions: R72P polymorphism in the TP53 gene was not associated to an increased risk of developing IDC in the population studied. This finding may be related to the great interracial and ethnic variation in our population deriving from frequent miscigenations and exposure to different environmental factors that are important in the evolution of breast cancer. There was also no association with clinical pathological characteristics.

Breast cancer is the most frequent malignant disease in the female population worldvide. About 10 % of all cases are of hereditary origin. The inactivation of tumor suppressor gene BRCA1 is the main genetic predisposing factor in breast cancer in the Czech Republic. Primarily, BRCA1 participates in DNA double strand break repair. Depending on cell cycle phase, the damage is repaired by homologous recombination or non-homologous end joining. Alternative splicing variants of BRCA1 are frequently detected during the genetic screening of high risk patients. The clinical significance of these variants is unknown. Understanding of the nature of breast cancer genetics is the critical factor for early diagnosis. Based on earlier studies from the Institute of Biochemistry and Experimental Oncology 1st Faculty of Medicine Charles University, two alternative splicing variants which were repeatedly detected in patients, were chosen for functional analysis. The aim of this work is to investigate the impact of alternative splicing variants BRCA1Δ5 and BRCA1Δ10 on DNA double strand breaks repair. Particular variants were over- expressed in the cells of model system. Activity of homologous recombination (HR) and non-homologous end joining (NHEJ) was scored by in vitro DNA repair assay. The cellular localization of...

Abstract Breast carcinoma is a heterogeneous disease with a prognosis that varies from excellent to very poor. Traditional tumour parameters and biological factors that are also predictive for treatment response are used in determining breast carcinoma prognosis and selecting appropriate treatment. Gelatinases MMP-2 and MMP-9 have been shown to associate with tumour progression. Their tissue inhibitors TIMP-1 and -2 are multifunctional molecules that have been suggested as prognostic markers in some previous reports. In the present work, the expression and prognostic value of gelatinases MMP-2 and MMP-9 and their tissue inhibitors TIMP-1 and -2 were assessed in primary breast carcinoma. The material consisted of a total of 416 patients. Tissue expression of TIMP-1 and -2 was analysed in a population of 203 patients using immunohistochemistry. Circulating gelatinases and their inhibitors were studied using ELISA in two different populations of 71 at preoperative state and 213 patients at pre- and postoperative state. High expression of TIMP-1 immunoreactive protein positively correlated with high histological grade of the tumour and associated with aggressive disease course in grade 2–3 subpopulation. High preoperative plasma TIMP-1 was prognostic for relapse in a modern patient series after a median follow-up time of 18 months. TIMP-1 as a continuous variable was prognostic in Cox regression univariate analysis, and was an independent prognostic variable superior to nodal status in multivariate analysis. High preoperative serum TIMP-1 was an independent prognostic variable for poor disease-specific survival, and TIMP-1 was found to maintain its prognostic value when assessed independently with different ELISA analyses, and was not very sensitive for preanalytical conditions. In addition, low circulating preoperative serum MMP-2 was observed to associate with high stage and positive nodal status in breast carcinoma. These results indicate that circulating TIMP-1 may be a potential new marker of worsened prognosis in breast carcinoma, although careful validation of assay platforms and identification of the sources of physiological variation are needed before it can be adopted into clinical decision-making.