Dissertations / Theses on the topic 'Breast Tumors Classification'

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) of the breast is a widely used non-invasive approach to gather information about the underlying physiology of breast tumors. Recent studies indicate that breast tumor heterogeneity may reflect the presence of different levels of cellular aggressiveness or habitats within the tumor. This heterogeneity has been correlated to the variations in the contrast enhancement patterns within the tumor apparent on gadolinium-enhanced DCE-MRI. Although pathological and qualitative (based on contrast enhancement patterns) studies suggest the presence of clini- cal and molecular predictive tumor sub-regions, this has not been fully investigated in the quantitative domain. The new era of cancer imaging emphasizes the use of Radiomics to provide in vivo quan- titative prognostic and predictive imaging biomarkers. Thus Radiomics focuses on apply- ing image analysis techniques to quantify tumor radiographic properties to create mineable databases from radiological images. In this research work, the Radiomics approach was ap- plied to develop a novel computer aided diagnosis (CAD) model for quantifying intratumor heterogeneity not only within the tumor as a whole, but also within tumor habitats with an intent to build predictive models in breast cancer. The process of building these predictive models started with 2-D tumor segmentation followed by habitat extraction (based on vari- ations in contrast patterns and geometry) and textural kinetic feature extraction to quantify habitat heterogeneity. A new correlation based random subspace ensemble framework was developed to evaluate the textural kinetics from the individual tumor habitats. This new CAD framework was applied to predict two clinical and prognostic factors: Axillary lymph node (ALN) metastases and Estrogen receptor (ER) status. An AUC of more than 0.8 was achieved for classifying breast tumors based on number of ALN involvement. The highest AUC of 0.91 was achieved for classifying tumors with no ALN metastases from tumors with 4 or more ALN metastases. For classifying tumors based on ER status the highest AUC of 0.87 was achieved. These results were acquired by utilizing the textural kinetic features from the tumor habitat with rapid delayed washout. The results presented in this work showed that the heterogeneity within the tumor habitats which showed rapid contrast washout in the delayed phase, correlated with aggressive cellular phenotypes. This work hypothesizes that successfully quantifying these prognostic factors will prove to be clinically significant as it can improve the diagnostic accuracy. This, in turn, will im- prove the breast cancer treatment paradigm by providing more tailored treatment regimens for aggressive tumors.

Computer-aided diagnosis for detection and classification of breast abnormalities on digital mammograms is an active area of. research. In the recent years, there have been many research developments in all aspects of the mammography. However, it still faces many challenges. The current detection accuracy of lesions such as mass and architectural distortion is considerably low. This research is focused on computerised texture analysis of mass and architectural distortion, and shape analysis of mass in digital mammograms. In texture analysis, we investigate fractal-based methods in texture characterisation of mammographic masses and architectural distortion.The individual ability of the different fractal-based features in the task of discriminating between abnormal lesion and normal breast parenchyma tissue are evaluated and compared using statistical analysis and receiver operating characteristics analysis.

Intravoxel Incoherent Motion Imaging (IVIM) is a non-invasive MR-imaging technique that enables the measurement of cellularity and vascularity using diffusion-weighted (DW)-imaging. IVIM has been applied to various cancer types including breast cancer, and is becoming more popular but lacks standardisation. The quantitative parameters; diffusion, D, perfusion fraction, f, and pseudo micro capillary diffusion, D* are thought to be correlated with tumour physiognomies such as proliferation, angiogenesis and heterogeneity. In Part 1 of this thesis, an optimised clinical b-value protocol is produced using a robust statistical method. This optimised protocol and various fitting methodologies are investigated in healthy volunteers, and then the most precise approach is applied in a clinical trial in patients following diagnosis of breast cancer, before treatment, to correlate IVIM parameters with breast cancer grade, histological type and molecular subtype with statistically significant results supporting IVIM’s potential as a non-invasive biomarker for malignancy. Monte Carlo simulations support this clinical application, where real data mean squared errors due to SNR limitations lie within simulated errors. A computed DW-imaging program is also presented to produce better quality images than acquired high b-value images as an adjunct to the optimised IVIM protocol. In Part 2 of this thesis, MR-guided Focused Ultrasound (MRgFUS) is explored as a means to create a pre-surgical template of thermally induced palpable markers to enable a surgeon to resect occult lesions and potentially reduce positive tumour margin status and local recurrence after breast conserving surgery. A surrogate animal model with pseudo lesion is presented, as well as a clinical tool to plan spot markers around a lesion as seen on MRI.

Dans cette thèse, je traite, d'un point de vue statistique, le sujet de la classification des tumeurs du cancer du sein triple négatif (TNBC). Je me concentre principalement sur l'utilisation de la stabilité des clusters pour sélectionner le nombre de groupes dans le clustering, la méthode généralement utilisée pour la classification des TNBC. L'objectif de cette méthode est d'obtenir une classification robuste, c'est-à-dire facilement reproductible sur des données similaires.Malgré sa popularité, on sait encore peu de choses sur la façon dont cette méthode fonctionne. Pour cette raison, je propose deux contributions méthodologiques importantes : (1) un package R, clustRstab}, qui permet d'estimer, de manière flexible, la stabilité d'un clustering avec différents paramètres. Ce package est accompagné d'une étude de simulation et d'une étude d'application qui examine sous quelles conditions cette méthode fonctionne. (2) Une version modifiée de la version Ajusté du Rand Index (ARI), un score populaire pour les comparaisons de clusters, étape cruciale pour estimer la stabilité d'un clustering. Je corrige ce score en le basant sur une hypothèse de distribution multinomiale qui lui permet de prendre en compte la dépendance entre les clusters et de faire des inférences statistiques. Ce ARI modifié (M ARI) est implémenté dans le package R aricode. Ces deux méthodes sont ensuite appliquées à une large cohorte de tumeurs TNBC et les résultats sont discutés en relation avec des résultats des classification du TNBC de la littérature
In this thesis, I treat the topic of classifying Triple Negative Breast Cancer (TNBC) tumors from a statistical point of view. After proposing a classification of TNBC based on proteins, I mainly focus on the use of cluster stability for selecting the number of groups in unsupervised clustering. Indeed, this is the method generally employed when classifying TNBC. The aim of this method is to obtain a classification that is robust, that is, easily replicable on similar data. This is measured by its sensibility to small changes, such as subsamplig of the dataset.Despite the popularity of this method, little is still known about how or when it works. For this reason, I propose two important methodological contributions, increasing the usability and interpretability of this method: (1) an R-package, clustRstab, that easily enables to estimate the stability of a clustering in different parameter settings. This package is accompanied by a simulation and an application study investigating when and how this method works. (2) A Modified version of the Adjusted Rand Index (ARI), a popular score for cluster comparisons which is a crucial step for estimating the stability of a clustering. I correct this score by basing it on a multinomial distribution hypothesis which enables it to take into account dependence between clusterings and conduct statistical inference. This Modified ARI (M ARI) is implemented in the R package texttt{aricode}.These two methods are then applied to a large cohort of TNBC tumors and the results are discussed in relation to earlier classification results of TNBC

Background: Using a nude rat model of site-specific metastatic bone disease (MBD), we developed a semiquantitative histological score for rapid assessment of lytic lesions in bone. This provides additional information to conventional histological measurement by clarifying the extent and location of metastatic infiltration and the tumor growth pattern. The score can also be used to assess the action of bisphosphonates on bone metastases. Materials and Methods: Male nude rats (n = 12 per group) were inoculated with the human breast cancer cell line MDA-MB-231 via the femoral artery. Following appearance of radiographically visible osteolytic lesions on day 18, the animals received phosphate-buffered saline (PBS; controls) or ibandronate (IBN, 10 µg P/kg) daily until day 30. Whole body radiographs were obtained on days 18 and 30, and osteolytic areas (OA) were determined by radiographic computer-based analysis (CBA). On day 30, MBD was assessed in both tibias using conventional histological CBA and the new scoring system. Results: Metastatic tumor area correlated with the total sum of the new score in both PBS- (r = 0.762) and IBN-treated animals (r = 0.951; p < 0.001). OA correlated well with the total sum in both groups (r = 0.845 and 0.854, respectively; p < 0.001). Conclusion: Significant reduction of bone marrow and cortical infiltration of tumor cells with IBN suggested local control of metastases.
Hintergrund: Mit Hilfe eines etablierten Tiermodells zur Erzeugung lokalisationsspezifischer Knochenmetastasen in der Nacktratte wurde ein semiquantitatives histologisches Graduierungssystem zur schnellen Bewertung osteolytischer Knochenmetastasen entwickelt. Das Graduierungssystem liefert hinsichtlich der Metastasenlokalisation, deren Ausmaß und Infiltrationsmuster wertvolle Zusatzinformationen zu den konventionellen histologischen Untersuchungsmethoden. Damit kann beispielsweise auch die pharmakologische Wirkung von Bisphosphonaten auf die Knochenmetastasierung beurteilt werden. Material und Methoden: Männlichen Nacktratten (n = 12 pro Gruppe) wurden Zellen der humanen Brustkrebszellinie MDA-MB-231 in die Oberschenkelarterie inokuliert. Ab dem Auftreten radiologisch erkennbarer Osteolysen 18 Tage nach Inokulation erhielten die Tiere bis zum Studienende (Tag 30) täglich entweder eine subkutane Applikation einer Phosphat-Puffer-Lösung (Kontrollgruppe) oder Ibandronat (IBN, 10 µg P/kg; Behandlungsgruppe). Konventionelle Röntgenaufnahmen wurden an den Tagen 18 und 30 nach Tumorinokulation angefertigt und die Osteolysenflächen mittels Computerauswertung bestimmt. Nach Studienende wurde der Metastasenbefall in beiden Tibiae sowohl konventionell histologisch als auch mittels des neuen Graduierungssystems ausgewertet. Ergebnisse: Die Metastasenfläche korrelierte mit der kummulativen Punktsumme des Graduierungssystems sowohl in der Kontrollgruppe (r = 0,762; p < 0,001) als auch in der Ibandronat- Gruppe (r = 0,951; p < 0,001). Ebenso war die Osteolysenfläche eng mit der Punktesumme in beiden Gruppen korreliert (r = 0,845 und 0,854; p < 0,001). Schlussfolgerung: Die signifikante Reduktion von Knochenmark- und Kortikalisbefall durch IBN deuten auf eine gute lokale Kontrolle des Metastasenwachstums hin.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Aim: To develop an immunomagnetic assay for the isolation of circulating tumor cells (CTCs) followed by the analysis of a multimarker panel, which will enable the characterization of these malignant cells with high accuracy. Patients and Methods: Peripheral blood (PB) was collected from 32 metastatic breast cancer patients and 42 negative controls. The antibodies BM7 and VU1D9 were used for immunomagnetic tumor cell enrichment. A real-time reverse transcription-polymerase chain reaction (RT-PCR) approach for the markers KRT19, SCGB2A2, MUC1, EPCAM, BIRC5 and ERBB2 was used for CTC detection and characterization. Results: The positivity rates for each marker were as follows: 46.9% for KRT19, 25.0% for SCGB2A2, 28.1% for MUC1, 28.1% for EPCAM, 21.9% for BIRC5, and 15.6% for ERBB2. After the creation of individualized cutoffs, the sensitivity and specificity of the combined marker gene panel increased to 56.3% and 100%, respectively. Interestingly, 27.0% of the HER2-negative tumor patients showed ERBB2 mRNA-positive CTCs. Conclusions: The described technique can be used to measure CTCs with great accuracy. The use of a multimarker panel for the characterization of CTCs may provide real-time information and be of great value in therapy monitoring.
Ziel: Entwicklung eines immunomagnetischen Verfahrens zur Isolierung zirkulierender Tumorzellen (CTCs) in Kombination mit einer molekularen Multimarkeranalyse für die hochspezifische Identifizierung maligner Zellen. Patientinnen und Methoden: Peripheres Blut (PB) von 32 Patientinnen mit metastasiertem Mammakarzinom und von 42 gesunden Kontrollen wurde für die immunomagnetische Tumorzellanreicherung mit den Antikörpern BM7 und VU1D9 genutzt. Eine Real-Time Reverse Transkription Polymerase-Kettenreaktion (RT-PCR)-Methodik mit den Markern KRT19, SCGB2A2, MUC1, EPCAM, BIRC5 und ERBB2 wurde für den CTC-Nachweis und die Tumorzellcharakterisierung entwickelt. Ergebnisse: Für die einzelnen Marker wurden die folgenden Positivitätsraten ermittelt: 46,9% für KRT19, 25,0% für SCGB2A2, 28,1% für MUC1, 28,1% für EPCAM, 21,9% für BIRC5 und 15,6% für ERBB2. Nach der Bestimmung individualisierter Cut-off-Werte ergab sich für den kombinierten Multimarkernachweis eine Sensitivität und Spezifität von 56,3% bzw. 100%. Bemerkenswert war der Befund, dass 27,0% der HER2-tumornegativen Patientinnen ERBB2-mRNA-positive CTCs aufwiesen. Schlussfolgerung: Die hier beschriebene Methodik bestimmt CTCs mit hoher Spezifität. Die molekulare Multimarkeranalyse liefert wertvolle Real-Time-Informationen für personalisierte Behandlungsmodalitäten.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Introdução: Os cânceres de mama subtipos Luminal A e B (HER2 negativo) podem apresentar prognóstico variável, a depender do índice de proliferação, avaliado pelo Ki67. As células malignas e as células estromais adjacentes (fibroblastos e células de resposta imune ) podem interagir tanto pelo contato célula a célula como por fatores secretados por elas, ambas influenciando no comportamento tumoral. Já foi demonstrado que as células estromais podem aumentar a proliferação das células do câncer da mama. Objetivo: Nosso objetivo foi avaliar o perfil de expressão gênica de células do estroma em câncer de mama luminal A e luminal B e analisar se este se correlaciona com o prognóstico da doença. Pacientes e Métodos/ Resultados: Amostras de tumores de 11 pacientes na pós menopausa foram analisadas, todas elas HER2 negativas. A expressão de Ki67 foi <= 10 % em 5 pacientes (luminal A) e >= 30 % em outras 6 amostras(Luminal B ). Células estromais foram microdissecadas para a extração de RNA, que posteriormente foi hibridizado na plataforma de microarray Agilent G485 -1A GE 8x60K. Após a normalização, 50 % dos genes com a maior variância foram selecionados para análise por SAM duas classes desemparelhado (software TMEV ) e aceitando FDR 14.1%, 35 sequências foram identificadas como diferencialmente expressas, incluindo 16 genes conhecidos, entre as células estromais das amostras de Luminal A versos Luminal B, todos mais expressos nas amostras B. Dentre as funções biológicas enriquecidas em genes diferencialmente expressos encontram-se regulação positiva do sistema imune, incluindo genes como ZAP70 (proteína quinase 70kDa associada a cadeia zeta (TCR)), CD38 (molécula CD38); UBASH3A (ubiquitina associada e SH3 domínio que contém A); PLA2G7 (fosfolipase A2, grupo VII (fator acetil ativador de plaquetas no plasma)); NCR3 (citotoxicidade natural, provocando receptor 3). Nosso próximo passo foi avaliar se a expressão de alguns genes selecionados estava associada com prognóstico de tumores luminais. Para tal selecionamos amostras de outro grupo de 89 pacientes com seguimento de pelo menos 5 anos, cujos tumores eram ER(+), HER2(-), para análise de expressão proteica em Tissue microarray. Caracterizamos os fibroblastos destas amostras com 3 marcadores de fibroblastos: actina de músculo liso (AML), S100A4 e caveolina-1 (CAV1) e analisamos a marcação da proteína ZAP70. Correlacionamos a expressão proteica de todos os marcadores com as características anatomopatológicas da amostra. Observamos que fibroblastos de todas as amostras de tumor de mama expressam AML, S100A4 e CAV1, em diferentes proporções, entretanto não detectamos diferença entre os tumores luminais A e B. Também não obsevamos diferença de expressão de AML, S100A4 e CAV1 em relação a grau histológico, comprometimento linfonodal e estadiamento clínico. Nestas amostras não detectamos expressão proteica de ZAP70 em fibroblastos tumorais. Conclusão: Houve expressão diferencial de 16 genes relacionados a processos imunes, todos eles mais expressos em células estromais de tumores Luminal B em relação a luminal A
Introduction: Luminal breast cancer subtypes A and B (HER2 negative) may present a variable prognosis, depending on tumor proliferation index, evaluated by Ki67 expression. Malignant cells and adjacent stromal cells (fibroblasts and immune response cells) may interact by both cell contact and secreted factors and influence tumor behavior. It was shown that stromal cells may enhance breast cancer cells proliferation. Objective: Our aim was to evaluate stromal cells gene expression profile in luminal A and luminal B tumors and to evaluate whether selected transcripts expressed in stromal cells may be associated with prognosis in breast cancer. Material/ Methods and Results: Hormone receptor positive tumor samples from 11 post menopausal patients were analyzed, all of them Her2 negative. Ki67 expression <= 10% (luminal A) was observed in five and Ki67 >= 30% (luminal B) in six samples. Stromal cells were microdissected for RNA extraction, which was hybridized in Agilent G485-1A GE 8x60K microarray platform. After normalization, 50% of the genes with the highest variance were selected for further analysis by two class unpaired SAM (TMEV software) and accepting FDR 14,1%, 35 sequences, including 16 known genes, were found differentially expressed between stromal cells from luminal A vs luminal B breast cancer samples, all of them more expressed in luminal B. Among biological functions enriched in genes found differentially expressed were positive regulation of immune system process, including genes as: ZAP70 (zeta-chain (TCR) associated protein kinase 70kDa); CD38 (CD38 molecule); UBASH3A (ubiquitin associated and SH3 domain containing A); PLA2G7 (phospholipase A2, group VII (platelet-activating factor acetylhydrolase, plasma); NCR3 (natural cytotoxicity triggering receptor 3). Our next step was evaluate whether expression of selected genes was associated with prognosis in another group of patients. Tumor samples from 89 patients with at least 5 years of follow up, all of them estrogen receptor positive and HER2 negative, were selected. Tissue microarray was prepared with stromal tumor compartment from paraffin embedded tumor samples. Fibroblasts were characterized for the expression of 3 fibroblasts markers (alfa-SMA, alpha smooth muscel actin; S100A4 and CAV1, caveolin 1), and ZAP70. Correlation of expression of these markers with prognostic variables was determined. Expression of alfa-SMA, S100A4 and CAV1 was detected in fibroblasts from all tumor samples in different proportions, however no differential expression was observed between luminal A and B tumors. Neither difference was detected on the expression of these proteins in relation with histological grade, lymph node involvement and clinical stage. Conclusion: A differential expression of 16 genes involved in immune process was found, all of them more expressed in fibroblasts from luminal B as compared with luminal A tumors

Changes in gene expression programs play a central role in cancer. Chromosomal aberrations such as deletions, duplications and translocations of DNA segments can lead to highly significant positive correlations of gene expression levels of neighboring genes. This should be utilized to improve the analysis of tumor expression profiles. Here, we develop a novel model class of autoregressive higher-order Hidden Markov Models (HMMs) that carefully exploit local data-dependent chromosomal dependencies to improve the identification of differentially expressed genes in tumor. Autoregressive higher-order HMMs overcome generally existing limitations of standard first-order HMMs in the modeling of dependencies between genes in close chromosomal proximity by the simultaneous usage of higher-order state-transitions and autoregressive emissions as novel model features. We apply autoregressive higher-order HMMs to the analysis of breast cancer and glioma gene expression data and perform in-depth model evaluation studies. We find that autoregressive higher-order HMMs clearly improve the identification of overexpressed genes with underlying gene copy number duplications in breast cancer in comparison to mixture models, standard first- and higher-order HMMs, and other related methods. The performance benefit is attributed to the simultaneous usage of higher-order state-transitions in combination with autoregressive emissions. This benefit could not be reached by using each of these two features independently. We also find that autoregressive higher-order HMMs are better able to identify differentially expressed genes in tumors independent of the underlying gene copy number status in comparison to the majority of related methods. This is further supported by the identification of well-known and of previously unreported hotspots of differential expression in glioblastomas demonstrating the efficacy of autoregressive higher-order HMMs for the analysis of individual tumor expression profiles. Moreover, we reveal interesting novel details of systematic alterations of gene expression levels in known cancer signaling pathways distinguishing oligodendrogliomas, astrocytomas and glioblastomas.

Brustkrebs ist die häufigste Krebstodesursache bei Frauen in den Industrienationen mit einer jährlich ansteigenden Neuerkrankungsrate (Senn und Niederberger 2002). Durch vorangegangene Untersuchungen wurde bereits deutlich, dass das Peptidhormon Relaxin unter in vitro Bedingungen maßgeblich zur Tumorprogression von Mammakarzinomen beiträgt (Binder et al. 2002). Die vorliegende Arbeit hat untersucht, ob Relaxin diese Wirkung auch in vivo auf Mammakarzinome ausübt. Relaxin ist ein multifunktionales Hormon. Es ist ein Aktivator verschiedenerWachstumsund Transkriptionsfaktoren (Samuel et al. 2007a) und nimmt eine Schlüsselfunktion im Bindegewebsstoffwechsel ein, indem es durch eine Steigerung der MMP-Expression zur bindegewebigen Erweichung führt (Unemori et al. 1996). Im Krebsgeschehen schafft das Peptidhormon damit die Voraussetzungen für Tumorwachstum und Metastasierung (Bingle et al. 2002). Für die Fragestellung der vorliegenden Arbeit wurde das Brustkrebsmodell der BalbneuT- Maus eingesetzt, die aufgrund der transgenen HER2-Überexpression spontan Mammakarzinome entwickelt. Es wurden 45 weibliche Tiere mit beginnendem Wachstum von Mammatumoren auf eine Relaxin- (n=22) und eine Kontrollgruppe (n=23) aufgeteilt. Den Tieren wurde über eine unter das Nackenfell implantierte osmotische Minipumpe (Fa. Alzet, Modell 2004; Kupertura, Kanada) im Falle der Relaxin-Gruppe Relaxin und im Falle der Kontrollgruppe isotone Natriumchloridlösung verabreicht. Danach wurden die Tiere 10-49 Tage beobachtet und daraufhin eingeschläfert. Es wurden die Tumoren, Biopsien von Leber, Lunge und Nieren sowie Blutproben entnommen. Um beurteilen zu können, ob die Tumoren der Relaxin-behandelten Tiere ein schnelleres Wachstum zeigten, wurden Tumorvolumina und -gewichte zu den unterschiedlichen Tötungszeitpunkten erfasst. Weiterhin wurden im Tumorgewebe immunhistochemisch der Proliferationsmarker Ki67, der Makrophagenmarker MAC 387, der Relaxinrezeptor RXFP1 sowie die Steroidhormonrezeptoren für 17!-Östradiol (ER) und Progesteron (PR) bestimmt. Zusätzlich wurde die RXFP1-spezifische mRNA molekularbiologisch im Tumorgewebe dargestellt. Außerdem wurden die peripheren Hormonkonzentrationen von Relaxin, 17!-Östradiol (E2) und Progesteron (P4) ermittelt. Die Ergebnisse der vorliegenden Arbeit konnten den Beweis erbringen, dass Relaxin auch in vivo dasWachstum von Mammakarzinomen unterstützt. Relaxin bewirkte im vorliegenden Experiment eine Rekrutierung von Tumor-assoziierten Makrophagen (TAMs) ins tumorumgebenden Bindegewebe. Dadurch erfolgte dort die Synthese verschiedener Faktoren und Enzyme, welche zur bindegewebigen Erweichung, Apoptosehemmung und zu einer gesteigerten Zellproliferation führten (Bingle et al. 2002; Devetzi et al. 2008). Weiterhin induzierte die exogene Relaxingabe eine vermehrte E2-Synthese, was sich ebenfalls wachstumsfördernd und apoptosehemmend auswirkte und somit die Tumorproliferation unterstützt hat (Catalano et al. 2009; Lewis-Wambi und Jordan 2009). Die Expression des RXFP1 im Tumorgewebe wurde durch Relaxin über eine gesteigerte E2- Synthese (Wilson et al. 2008) gefördert, ebenso wie die Expression des ER. Weiterhin führte Relaxin zu einer gesteigerten P4-Synthese und zur gesteigerten Expression des PR im Tumorgewebe über einen derzeit noch unbekannten Mechanismus. Aufgrund der maßgeblichen Bedeutung des Peptidhormons für das Progressionsverhalten von Mammakarzinomen kann die Bestimmung der Relaxinblutspiegel bei Brustkrebspatientinnen deshalb in Zukunft ein wichtiges Hilfsmittel bei der Wahl der richtigen Therapie und bei der Prognosebeurteilung werden.

Циљ ове студије је да се упореди дијагностичкa значајност 2Д и 2Д+3Д мамографије у детектовању тумора дојке. Испитивали смо 864 дојки у 740 пацијенткиња. Студија је спроведена у току рутинског рада у Центру за имиџинг дијагностику Институту за онкологију Војводине. 2Д + 3Д мамографија су начињене појединачно или у истом акту снимања као комбо опција на Selenia Dimensions апарату произвођача фирме Холоџик. Радиолошки извешатаји су класификовани у категорије 1-5 према АЦР БИ РAДС-у. Патохистолошка верификација је вршена у свих суспектних промена или у току њиховог праћења . Све пацијенткиње са уредним налазом или мамографски уочених бенигних промена су радиолошки праћене најкраће током 2 године. Уочено је 103 малигне лезије у дојкама класификованих као БИ РАДС 4, 5 на дигиталној мамографији и у 22 дојке чије су промене класификоване као БИ РАДС 1-3, током праћења или прегледа дојки помоћу других модалитета. На 2Д + 3Д мамографији малигните је потврђен у 125 дојке од којих је 118 класификовано као БИ РАДС 4,5 и у 7 дојки чује су промене категорисане у БИРАДС 1-3. Постоји статистички значајна разлика у дистрибуцији малигних налаза у односу на подгрупе Студија је показала 20% лажно негативних налаза на 2Д, а 5,6% на 2Д + 3Д модалитету прегледа дојки. Осетљивост у откривању рака у овој студији износи 82,4% на 2Д и 94,4% на 2Д+3Д методи прегледа, док је специфичност 90,5% и 92,0%, респективно. ППВ је већа за 2Д + 3Д технику прегледа , износи 66,7%, као и негативна предиктивна вредност која износи 99,0%. У 172 случаја (19, 9%) налази 2Д мамографије се не уочавају на 3Д техници прегледа и сматрају се последицом структурне или анатомске „буке“. Већина не -сталних налаза (85%) је класификовано као фокална асиметрија. У овој студији 500 дојки је класификовано према АЦР структури у масне (АЦР 1) или дифузне фибро-гландуларне (АЦР 2), а преосталих 264 је било хетеродензно (АЦР 3) и значајно дензно (АЦР 4). Статистички значајна разлика није показана приликом дистрибуције малигних налаза у поређењу са подгрупама дојки начињеним према њиховој густини - складу са правилима АЦР-а. Укупна тачност теста износи 89,4% за 2Д и 92,4% за 2Д + 3Д мамографију. Предиктивне вредности добијене за 2Д + 3Д мамографију су боље од оних које се односе само на 2Д мамографију, што је резултат њене веће осетљивости и шире могућности карактеризације промена. Варијабилност у интерпретацији налаза међу два радиолога је је ниска, показано је слагање у интершпретавцији евалуираних мамограма у 94.1% случајева.
Cilj ove studije je da se uporedi dijagnostička značajnost 2D i 2D+3D mamografije u detektovanju tumora dojke. Ispitivali smo 864 dojki u 740 pacijentkinja. Studija je sprovedena u toku rutinskog rada u Centru za imidžing dijagnostiku Institutu za onkologiju Vojvodine. 2D + 3D mamografija su načinjene pojedinačno ili u istom aktu snimanja kao kombo opcija na Selenia Dimensions aparatu proizvođača firme Holodžik. Radiološki izvešataji su klasifikovani u kategorije 1-5 prema ACR BI RADS-u. Patohistološka verifikacija je vršena u svih suspektnih promena ili u toku njihovog praćenja . Sve pacijentkinje sa urednim nalazom ili mamografski uočenih benignih promena su radiološki praćene najkraće tokom 2 godine. Uočeno je 103 maligne lezije u dojkama klasifikovanih kao BI RADS 4, 5 na digitalnoj mamografiji i u 22 dojke čije su promene klasifikovane kao BI RADS 1-3, tokom praćenja ili pregleda dojki pomoću drugih modaliteta. Na 2D + 3D mamografiji malignite je potvrđen u 125 dojke od kojih je 118 klasifikovano kao BI RADS 4,5 i u 7 dojki čuje su promene kategorisane u BIRADS 1-3. Postoji statistički značajna razlika u distribuciji malignih nalaza u odnosu na podgrupe Studija je pokazala 20% lažno negativnih nalaza na 2D, a 5,6% na 2D + 3D modalitetu pregleda dojki. Osetljivost u otkrivanju raka u ovoj studiji iznosi 82,4% na 2D i 94,4% na 2D+3D metodi pregleda, dok je specifičnost 90,5% i 92,0%, respektivno. PPV je veća za 2D + 3D tehniku pregleda , iznosi 66,7%, kao i negativna prediktivna vrednost koja iznosi 99,0%. U 172 slučaja (19, 9%) nalazi 2D mamografije se ne uočavaju na 3D tehnici pregleda i smatraju se posledicom strukturne ili anatomske „buke“. Većina ne -stalnih nalaza (85%) je klasifikovano kao fokalna asimetrija. U ovoj studiji 500 dojki je klasifikovano prema ACR strukturi u masne (ACR 1) ili difuzne fibro-glandularne (ACR 2), a preostalih 264 je bilo heterodenzno (ACR 3) i značajno denzno (ACR 4). Statistički značajna razlika nije pokazana prilikom distribucije malignih nalaza u poređenju sa podgrupama dojki načinjenim prema njihovoj gustini - skladu sa pravilima ACR-a. Ukupna tačnost testa iznosi 89,4% za 2D i 92,4% za 2D + 3D mamografiju. Prediktivne vrednosti dobijene za 2D + 3D mamografiju su bolje od onih koje se odnose samo na 2D mamografiju, što je rezultat njene veće osetljivosti i šire mogućnosti karakterizacije promena. Varijabilnost u interpretaciji nalaza među dva radiologa je je niska, pokazano je slaganje u interšpretavciji evaluiranih mamograma u 94.1% slučajeva.
The aim of this study was to compare diagnostic importance of 2D and 2D+3D diagnostic mammography in breast tumor detection. We evaluated 864 breasts in 740 patients. Study was performed in Diagnostic Imaging Center at Oncology Institute of Vojvodina. 2D+3D mammography were performed during single procedure or via combo option at Selenia Dimensions unit, Hologic, BE. Radiological findings were classified in categories 1-5 according to ACR BIRADS. Pathohistologic verification was obtained in all suspicious findings or after follow up studies. All other patients with mammographic normal findings or benign findings were fallowed up during 2 years period or longer. We detected malignant lesions in 103 breasts classified as BIRADS 4,5 at digital mammography, and in 22 breasts classified as BIRADS 1-3 after followed up or diagnosed by other imaging modalities. At 2D+3D mammography malignancy was confirmed in 125 breasts, 118 classified as BIRADS 4,5 and in 7 breasts classified as BIRADS 1-3. There is statistically significant difference (p<0.001) in distribution of malignant findings compared to the subgroups classified according to 2D mammography. There was 20% false negative findings on 2D, and 5.6% on 2D+3D modality. Sensitivity in cancer detection in this study is 82.4% and 94.4% for 2D and 2D+3D mammography, while specificity is 90.5% and 92.0%, respectively. PPV is higher for 2D+3D technique (66.7%), as well as negative predictive value (99.0%). In 172 cases (19, 9%) 2D mammography findings did not persist on 3D mammography and were considered as structural or anatomical noise. The majority of the non-persistent findings (85%) were classified as asymmetric focal density. In this study 500 breast were classified according to ACR as fatty (ACR 1) or scattered fibroglandular densities (ACR 2), and the remaining 264 had heterogeneously (ACR 3) and extremely dense breasts (ACR 4). Statistically significant difference (p<0.001) was not shown in distribution of malignant findings compared to the subgroups of density structure according to ACR. Overall accuracy of the test was 89.4% and 92.4% for 2D and 2D+3D mammography, respectively. Predictive values obtained in 2D+ 3D mammography are better than those for 2D mammography alone, as a result of its higher sensitivity and better possibility of lesion characterization. Interobserver variability is low, there is an agreement between two radiologist between two radiologic interpretations in 94.1% cases.

Hintergrund Die Strahlentherapie ist neben der radikalen Prostatektomie eine Standardtherapie zur Behandlung von Prostatatumoren und führt zu sehr guten Ergebnissen für die lokale Tumorkontrolle und für das Überleben. Allerdings ist, wie bei der Operation auch, dabei das Risiko eines Rezidivs für fortgeschrittene Tumoren im Gegensatz zu Tumoren in früheren Stadien relativ hoch. Daher besteht eine hohe Dringlichkeit zur Verbesserung der Strahlentherapie vor allem bei fortgeschrittenen Tumoren. Ein Ansatz hierfür ist die Kombination der Bestrahlung mit molekularen Therapien. Ziel dabei ist es, bestimmte Zielproteine zu blockieren, um die Strahlensensibilität der Prostatakarzinomzellen zu erhöhen. Ein potentielles Target könnte hierbei die Blockade des VEGF-C/NRP-2/Akt-Signalwegs (VEGF-C – vascular endothelial growth factor C; NRP-2 – Neuropilin 2; Akt – Proteinkinase B) sein. Im Prostatakarzinom sind die Konzentrationen von VEGF-C und NRP-2 im Vergleich zu normalen Prostatazellen erhöht. Aus Untersuchungen ist bekannt, dass beide Proteine eine progressive Wirkung auf die Tumorgenese haben. In Vorarbeiten zeigen Muders et al. (2009) zudem eine Aktivierung von Akt über die VEGF-C/NRP-2-Achse und eine darüber vermittelte Resistenz gegenüber oxidativem Stress durch H2O2. Akt wirkt in verschiedenen Tumorentitäten außerdem protektiv gegenüber Bestrahlung. Es besteht die Annahme, dass dies auch für Prostatakarzinomzellen gilt. Zielstellung Im Rahmen dieser Arbeit wurde untersucht, ob und über welchen Mechanismus VEGF-C, NRP-2 und Akt die Strahlenresistenz in Prostatakarzinomzelllinien beeinflussen. Methoden Es wurden in vitro- und in vivo-Experimente in den humanen Prostatakarzinomzelllinen PC-3, DU145, LNCaP sowie in PC-3-Xenografts durchgeführt. Der Einfluss von VEGF-C und NRP-2 auf die Strahlenresistenz wurde in vitro nach Herunterregulierung der entsprechenden Gene mittels siRNA beziehungsweise nach Supplementierung mit humanem rekombinanten VEGF-C in Koloniebildungsassays untersucht. Zur Ermittlung des Einflusses von VEGF-C und von NRP-2 auf mögliche Zellüberlebensmechanismen wurden der autophagische Flux nach Blockade der Autophagie mit Bafilomycin A1 mittels Western Blot, die DNA-Doppelstrangbruch-Reparatur mittels Quantifizierung der γH2AX Foci sowie die Zellzyklusverteilung mittels Durchflusszytometrie untersucht. Die Signalweiterleitung von VEGF-C über Akt sowie, als weitere Möglichkeit, die Signalweiterleitung über ERK1/2 wurden nach siRNA-Transfektion mit und ohne Bestrahlung mittels Western Blot geprüft. Weitere Versuche zu Akt erfolgten in vitro und in vivo mit dem PI3K/Akt-Inhibitor Nelfinavir in PC-3-Zellen. Der in vitro Effekt von Nelfinavir auf die Strahlenresistenz wurde dabei mithilfe eines Koloniebildungsassays nach Behandlung der Zellen mit 10 µM Nelfinavir getestet. In vivo wurde die Wirkung von Nelfinavir ohne sowie in Kombination mit Bestrahlung in PC-3-Xenografts in Nacktmäusen untersucht. Für die Bestimmung der Tumorwachstumszeit wurden die Mäuse mit 80 mg Nelfinavir/kg Körpergewicht 30 mal innerhalb von 6 Wochen behandelt. In einem weiteren Versuch wurde die lokale Tumorkontrolle bei gleichzeitiger fraktionierter Bestrahlung mit Gesamtdosen von 30 bis 120 Gy und einer Nachbeobachtungszeit von 180 Tagen bestimmt. Ergebnisse Die Untersuchungen zur Strahlenresistenz über den VEGF-C/NRP-2/Akt-Signalweg haben ergeben, dass in den drei Prostatakarzinomzelllinien PC-3, DU145 und LNCaP VEGF-C signifikant Strahlenresistenz vermittelt. Für NRP-2 hingegen wurde festgestellt, dass es in Abhängigkeit von der Zelllinie entweder zur Strahlenresistenz (DU145) oder zur Strahlensensibilisierung (PC-3) führt. Weiterhin wurde nachgewiesen, dass durch VEGF-C in PC-3 und DU145 weder über Akt noch über ERK1/2 Strahlenresistenz vermittelt wird. Die Versuche zu Strahlenresistenz vermittelnden Mechanismen ergaben, dass VEGF-C in unbestrahlten PC-3-Zellen die Autophagie fördert, NRP-2 jedoch nicht. Unter Bestrahlung war ein Effekt von VEGF-C und NRP-2 auf die Autophagie nicht reproduzierbar nachweisbar. Ein weiterer Versuch hat gezeigt, dass in PC-3 Autophagie keinen Einfluss auf das klonogene Überleben nach Bestrahlung hat. Außerdem wurde festgestellt, dass VEGF-C in PC-3 die DNA-Doppelstrangbruch-Reparatur nicht beeinflusst. Darüber hinaus wurde nachgewiesen, dass eine Verminderung des VEGF-C-Gehalts in PC-3 zum G2/M-Arrest führt. In DU145 konnte jedoch kein Effekt beobachtet werden. In den Untersuchungen zum Einfluss von Akt auf die Strahlenresistenz unabhängig von VEGF-C und NRP-2 wirkte Nelfinavir inhibierend auf die Akt-Phosphorylierung am Ser473 und beeinflusste das klonogene Überleben von PC-3-Zellen minimal. In PC-3-Xenografts führte Nelfinavir zu keiner Tumorwachstumsverzögerung und wirkte in vitro und in vivo nicht strahlensensibilisierend. Schlussfolgerung In den Versuchen konnte gezeigt werden, dass VEGF-C in Prostatakarzinomzellen Strahlenresistenz vermittelt. Diese Erkenntnis könnte als ein Forschungsansatz zur Entwicklung einer kombinierten Therapie aus VEGF-C-Blockade und Bestrahlung dienen. Ein potentieller Mechanismus, über den VEGF-C die Strahlenresistenz vermittelt, ist, in Abhängigkeit von der Zelllinie, die Aufhebung des G2/M-Arrests. NRP-2 wirkt in der Vermittlung von Strahlenresistenz beziehungsweise sensibilität je nach Zelllinie unterschiedlich. Hierzu sollten weitere Untersuchungen bezüglich möglicher Interaktionen innerhalb anderer Signalwege mit strahlensensibilisierendem Einfluss erfolgen. Innerhalb des untersuchten Signalwegs konnte weiterhin festgestellt werden, dass VEGF-C Strahlenresistenz nicht über Akt vermittelt. Die vorliegende Arbeit enthält die erste Studie sowohl zur Untersuchung des Einflusses von Nelfinavir in Kombination mit Bestrahlung auf das Überleben von Prostatakarzinomzellen in vitro als auch auf die Tumorwachstumszeit und die lokale Tumorkontrolle in vivo. Hierin konnte keine strahlensensibilisierende Wirkung von Nelfinavir nachgewiesen werden. Da Nelfinavir in Zellen anderer Tumorentitäten strahlensensibilisierend wirkt und außerdem bekannt ist, dass es in eine Reihe von Signalwegen eingreift, die das Zellüberleben fördern oder hemmen, sollte weiter geklärt werden, ob Tumorzellen mit einem bestimmten genetischen Profil besser auf die Behandlung mit Nelfinavir ansprechen
Background In addition to radical prostatectomy, radiotherapy is a standard therapy for the treatment of prostate tumours and leads to good results for local tumour control and survival. However, as with the resection, the risk of recurrence for advanced tumours is relatively high compared to tumours in earlier stages. Therefore, there is a high urgency to improve radiotherapy especially for advanced stages. One approach is the combination of irradiation with molecular therapies. The aim is to block certain target proteins to increase the radiosensitivity of the prostate carcinoma cells. A potential target could be the blockade of the VEGF-C/NRP-2/Akt signalling pathway (VEGF-C – vascular endothelial growth factor C; NRP-2 – neuropilin 2; Akt – protein kinase B). In prostate cancer the concentrations of VEGF-C and NRP-2 are increased compared to normal prostate cells. Studies have shown that both proteins have a progressive effect on tumourigenesis. In preliminary work Muders et al. (2009) also showed the activation of Akt via the VEGF-C/NRP-2 axis and a resistance to H2O2 induced oxidative stress. Akt also has a protective effect against irradiation in various tumour entities. It is assumed that this also applies to prostate carcinoma cells. Aim of the study Within the framework of this thesis, it was investigated whether and via which mechanism VEGF-C, NRP-2, and Akt affect the radioresistance in prostate carcinoma cell lines. Methods In vitro and in vivo experiments were performed in the human prostate carcinoma cell lines PC-3, DU145, LNCaP, as well as in PC-3 xenografts. The influence of VEGF-C and NRP-2 on the radioresistance was examined in vitro after knock down of the corresponding genes using siRNA or after supplementation with human recombinant VEGF-C in colony formation assays. In order to determine the influence of VEGF-C and NRP-2 on possible cell survival mechanisms, the autophagic flux was examined after the blockade of autophagy with bafilomycin A1 using western blot, the DNA double strand break repair by quantification of the γH2AX foci, and the cell cycle distribution by flow cytometry. The signal transduction of VEGF-C via Akt as well as, as a further possibility, the signal transduction via ERK1/2 were tested after siRNA transfection with and without irradiation using western blot. Further experiments on Akt were performed in vitro and in vivo with the PI3K/Akt inhibitor nelfinavir in PC-3 cells. The in vitro effect of nelfinavir on radioresistance was tested using a colony formation assay after treatment of the cells with 10 μM nelfinavir. In vivo, the effect of nelfinavir without and in combination with irradiation in PC-3 xenografts was investigated in nude mice. For the determination of the tumour growth time, the mice were treated with 80 mg nelfinavir/kg body weight 30 times within 6 weeks. In a further experiment, the local tumour control was determined with simultaneous fractionated irradiation with total doses of 30 to 120 Gy and a follow-up time of 180 days. Results The investigations on radioresistance via the VEGF-C/NRP-2/Akt signalling pathway showed that in the three prostate carcinoma cell lines PC-3, DU145, and LNCaP VEGF-C significantly mediates radioresistance. For NRP-2 however, it was found that, depending on the cell line, it either leads to radioresistance (DU145) or radiosensitization (PC-3). Further, it was shown that in PC-3 and DU145 VEGF-C does not mediate radioresistance via Akt or ERK1/2. The experiments on radioresistance mediating mechanisms revealed that VEGF-C promotes autophagy in untreated PC-3 cells, but NRP-2 does not. Under irradiation, an effect of VEGF-C and NRP-2 on autophagy could not be detected reproducibly. A further experiment has shown that in PC-3 autophagy has no influence on the clonogenic survival after irradiation. In addition, it was found that VEGF-C does not affect the DNA double strand break repair in PC-3. Furthermore, it was shown that a reduction in the VEGF-C content leads to a G2/M arrest in PC-3. However, no effect could be observed in DU145. In studies regarding the influence of Akt on radioresistance independent of VEGF-C and NRP-2, nelfinavir inhibited Akt phosphorylation at Ser473 and minimally affected the clonogenic survival of PC-3 cells. In PC-3 xenografts, nelfinavir did not lead to any tumour growth delay and did not have a radiosensitizing effect in vitro or in vivo. Conclusion In the experiments, it was shown that VEGF-C mediates radioresistance in prostate cancer cells. This finding could serve as a research approach for the development of a combined therapy of a VEGF-C blockade and irradiation. A potential mechanism by which VEGF-C mediates radioresistance is the reverse of the G2/M arrest, depending on the cell line. NRP-2 acts differently in the mediation of radioresistance or radiosensitivity, depending on the cell line. On this, further investigations should be carried out with regard to possible interactions within other signalling pathways with a radiosensitizing influence. Within the investigated signalling pathway, it was further shown that VEGF-C does not mediate radioresistance via Akt. The present work contains the first study examining the effect of nelfinavir in combination with irradiation on prostate cancer cell survival in vitro as well as on growth time and local tumour control in vivo. Herein no radiosensitizing effects of nelfinavir could be detected. Since nelfinavir radiosensitizes cells of other tumour entities and is also known to interfere with a series of signalling pathways that promote or inhibit cell survival, it should be clarified whether tumour cells with a particular genetic profile are more responsive to treatment with nelfinavir

Medical Microwave Imaging (MMI) has been studied in the past years to develop techniques to detect breast cancer at the earliest stages of development. Particularly, ultra-wideband (UWB) micro-wave radar imaging systems can detect and classify tumors as benign or malignant since this technique yields information about the size and shape of tumors. In this study we used this technology to classify tumors. The primary goal of this dissertation is two-folded. First, producing breast tumor numerical mod-els and using them in 2D MMI simulations that recreate the conditions of a UWB microwave radar imaging system. The breast tumor numerical produced resemble real tumor morphologies since they are made from breast MRI exams segmentations. Second, the data of the backscattered UWB microwave signals produced by the MMI simulations was used to classify tumors according to their size and histol-ogy, which is relevant to assess potential of UWB microwave radar imaging systems as a reliable alter-native method for the classification of breast tumors in the field of Medical Microwave Imaging. The Classification Algorithms used in this work were Pseudo Linear Discriminant Analysis (Pseudo-LDA), Pseudo Quadratic Discriminant Analysis (pseudo-QDA), and k-Nearest Neighbors (KNN), alongside with a feature extraction algorithm – Principal Component Analysis (PCA).
A Imagem Médica por Microondas (do inglês, MMI) tem sido estudada nos últimos anos de forma a desenvolver técnicas de deteção do cancro da mama nas primeiras fases de desenvolvimento. Em particular, os sistemas de imagem de radar por microondas em banda ultralarga (do inglês UWB) podem detetar e classificar os tumores como benignos ou malignos, uma vez que esta técnica produz informação sobre o tamanho e a forma dos tumores. Neste estudo, utilizámos esta tecnologia para classificar os tumores. A dissertação tem dois objetivos principais. Primeiro, produzir fantomas de tumores mamários e utilizá-los em simulações de MMI em 2D que recriam as condições de um sistema de imagem de radar por microondas UWB. Os fantomas numéricos de tumores mamários produzidos possuem morfologias semelhantes a tumores reais, uma vez que são feitos a partir de segmentações de exames de ressonância magnética da mama. Em segundo lugar, as reflexões dos sinais de microondas UWB produzidos pelas simulações de MMI foram utilizados para classificar tumores de acordo com o seu tamanho e histologia, o que é relevante para avaliar o potencial dos sistemas de imagem de radar por microondas UWB como um método alternativo e fiável para a classificação de tumores mamários no campo da MMI. Os Algo-ritmos de Classificação utilizados neste trabalho foram a Pseudo Linear Discriminant Analysis (Pseudo-LDA), Pseudo Quadratic Discriminant Analysis (pseudo-QDA), e a K-Nearest Neighbors (KNN), jun-tamente com um algoritmo de extração de features - Análise de Componentes Principais (do inglês PCA).

碩士
長庚大學
資訊管理研究所
94
According to the statistics from the Department of Health, breast cancer is still on the top-5 list of the cause of death. Hence, regular health check and set up of a computer-aided diagnosis system is of great urgency. Owing to the shape of a benign tumor is quite different from a malignant tumor and is relatively stable that comparing with conventional features like texture, so it is not affected with different type of scanners. In this thesis, we classify a tumor according to its shape feature. The result shows that the classification result is good and the accuracy is 89.05%. Furthermore, we use a support vector machine (SVM) as the classifier. A traditional SVM is sensitive to the outlier in the samples. It affects the location of the decision hyper-plane and decreases the accuracy. Hence, we propose an effective method to solve this problem and will improve the performance of the SVM. The experiment shows that the proposed method successfully reduces the influence of the outliers and promotes the classification accuracy of a SVM form 89.05% to 91.43%.

Os tumores mamários caninos (TMC) são considerados a neoplasia mais frequente em cadelas. Diferentes estudos indicam que cerca de 50% das cadelas com tumores mamários apresentam tumores mamários benignos e 50% manifestem tumores malignos. A evolução clínica dos tumores malignos é muito variável, sendo que o tempo livre de doença e o tempo de sobrevida podem variar entre poucos meses a vários anos. Os principais fatores de risco associados aos TMC são a idade, a raça, a exposição a hormonas gonadais ou seus análogos, a idade de realização da ovariohisterectomia e a obesidade. A classificação tumor-linfonodo-metástases (TNM) e o estadio clínico, a classificação histológica, o grau histológico de malignidade de Nottingham, o tipo de cirurgia e determinados biomarcadores séricos e tecidulares são atualmente considerados os fatores de prognóstico mais importantes. Nos últimos anos, os biomarcadores séricos e tecidulares têm sido cada vez mais estudados, uma vez que poderão contribuir de forma decisiva para a abordagem ao TMC. Os marcadores tecidulares mais estudados são os recetores das hormonas sexuais– recetores de estrogénio e de progesterona, o recetor do fator de crescimento epidermal humano 2 (HER-2/neu), o gene de suscetibilidade ao cancro mamário 1 (BRCA1), o p53 e a E-caderina. Os marcadores séricos que poderão ter utilidade ou valor de prognóstico incluem o antigénio cancerígeno 15-3 (CA15-3), a interleucina 8 e 10, a timidina quinase (TK1), a lactato desidrogenase (LDH), a mamoblobina B, a mucina 1 (MUC1) e as sequências CAN SINE (elementos nucleares intercalados simples). Existem também alguns estudos sobre os marcadores de proliferação – regiões organizadoras nucleolares argirófilas (AgNOR), antigénio nuclear de proliferação celular (PCNA) e Ki-67. Foram também publicados estudos sobre a enzima COX-2, os inibidores do ciclo dependente da quinase – p16, p21 e p27, o gene TP53, e a glicoproteína antigénio carcino-embrionário (CEA). Os fatores de prognóstico dos tumores mamários caninos apresentam-se de grande relevância devido à elevada incidência destes tumores e pelas implicações clínicas associadas à sua utilização. Adicionalmente, os tumores mamários da cadela têm também sido usados como modelos biológicos espontâneos dos tumores mamários em humanos. O presente trabalho pretendeu fazer uma revisão sobre os fatores de prognóstico existentes, tendo em conta a sua utilidade clínica documentada.
Canine mammary tumors (TMC) are considered the most frequent neoplasia in female dogs. Different studies indicate that about 50% of bitches with breast tumors have benign breast tumors, and 50% have malignant tumors. The clinical evolution of malignant tumors varies widely, with disease-free time and survival time ranging from a few months to several years. The main risk factors associated with CMD are age, breed, exposure to gonad hormones or analogues, age of ovariohysterectomy and obesity. The tumor-nodes-metastasis (TNM) classification, the clinical stage and histological classification, the histological grade of Nottingham's malignancy, the type of surgery and serum and tissue biomarkers are currently considered the most important prognostic factors. In recent years, serum and tissue biomarkers have been increasingly studied as they may contribute decisively to the approach to TMC. The most studied tissue markers are the sex hormone receptors- estrogen and progesterone receptors; the expression of human epidermal growth factor receptor 2 (HER-2 / neu); the breast cancer susceptibility gene 1 (BRCA1); p53 and E-cadherin. Serum markers that may have utility or prognostic value are carcinoembryonic antigen (CA15-3), Interleukin 8 and 10, TK1, lactate dehydrogenase (LDH), Mamoblobin B, mucin 1 (MUC1), and CAN SINE (single intercalated nuclear elements) sequences. There are also some studies on proliferation markers – argyrophilic nucleolar organizer region (AgNOR), proliferating cell nuclear antigen (PCNA) and Ki-67; the COX-2 enzyme; the kinase dependent cycle inhibitors - p16, p21 and p27; the TP53 gene; and the carcinoembryonic antigen (CEA) glycoprotein. This subject is of high relevance due to the high incidence of this type of tumor in dogs, and the clinical implications it may present. In addition, bitch mammary tumors have also been used as spontaneous biological models of mammary tumors in humans. The present study is a review of existing prognostic factors, taking into account the documented utility of these as a prognostic factor, since they have an influence on the treatment of neoplasia.

An 53 Hündinnen aus dem Patientengut der Klinik für Kleintiere der Universität Leipzig, die mit Umfangsvermehrungen der Mamma vorgestellt und anschließend in der Klinik für Kleintiere operiert wurden, wurde präoperativ eine sonographische Untersuchung der Mamma durchgeführt. Darüber hinaus wurden die Mammarkomplexe von acht tragenden und einer laktierenden Hündin mit dieser Technik untersucht. Ziel war es, Kriterien zur Einschätzung der Dignität der Tumoren mit Hilfe dieser nichtinvasiven Methode zu erarbeiten. Es sollten die Fragen geklärt werden, ob mit Hilfe der hochauflösenden Sonographie eine Aussage über Gut- oder Bösartigkeit eines Herdes möglich ist und ob dabei dieselben Kriterien entscheidend sind, die in der Humanmedizin eine zuverlässige Differenzierung erlauben. Außerdem sollte überprüft werden, welchen Beitrag die farbkodierte Duplexsonographie oder Resistance- und Pulsatilitätsindex zur Charakterisierung von Mammatumoren der Hündin leisten. Die Gesamtzahl der in die Studie eingehenden Komplexe beträgt 114. Die sonographischen Untersuchungen erfolgten mit einem 14 MHz Matrix-Linearschallkopf. Bei 70 der 114 untersuchten Lokalisationen erfolgte zusätzlich zur B-Mode-Untersuchung eine Untersuchung mit der farbkodierten Duplexsonographie. Konnten mit Hilfe dieser Methode Gefäße in der Umfangsvermehrung nachgewiesen werden, wurde in 47 von 70 Fäl-len zusätzlich der PW-Doppler eingesetzt, um Flussspektren aus den dargestellten Gefäßen abzuleiten. Aus diesen wurden Resistance-Index und Pulsatilitätsindex bestimmt. Bei der retrospektiven Auswertung der Grauwertbilder aus der B-Mode-Untersuchung wurde für jeden Komplex die Ausprägung von 12 Parametern beurteilt. Die Bilder aus der farbkodierten Duplexsonographie lieferten zusätzlich Informationen zu Gefäßzahl, Gefäßdurchmesser und Gefäßverteilung innerhalb eines Tumors. Die Exstirpate wurden pathohistologisch untersucht. Die aus der Gewebetypisierung entsprechend der WHO-Klassifikation resultierenden Gruppen sind so klein, dass nur eine deskriptive statistische Auswertung möglich war. Es erfolgte die Zusammenfassung unterschiedlicher Gewebetypen zu den Gruppen der „malignen“ bzw. „benignen“ Tumoren. Für Malignität sprechen eine unregelmäßige Randkontur (32 von 61 malignen, 4 von 48 benignen Lokalisationen), eine Schallverstärkung (36/61 malignen, 9/48 benignen Lokalisationen) oder –auslöschung (8/61 malignen, 0 /48 benignen Lokalisationen) hinter dem Tumor, Verkalkungen (20/61 malignen, 6/48 benignen Lokalisationen) sowie ein unregelmäßiger Durchmesser der Tumorgefäße (25/61 malignen, 12/48 benignen Lokalisationen). Meist gutartig sind Umfangsvermehrungen der Mamma, denen sonographisch eine klare Abgrenzung zum umgebenden Gewebe fehlt (15/61 malignen, 36/48 benignen Lokalisationen). Außerdem solche mit indifferentem retroläsionalem Schallverhalten (17/61 malignen, 39/48 benignen Lokalisationen). Kombiniert man mehrere der Parameter miteinander, ist die resultierende Teilmenge der betreffenden Läsionen kleiner, die Aussagekraft höher. Für Bösartigkeit spricht beispielsweise eine Kombination von Verkalkung und unregelmäßiger Randkontur (13 von 61 malignen, 1 von 48 benignen Lokalisationen), Verkalkung und echodichtem Randsaum („deutlich“ oder „fraglich“; 9/61 malignen, 0/48 benignen Lokalisationen) sowie mittlerer Echodichte und retroläsionaler Schallverstärkung (21/61 malignen, 6/48 benignen Lokalisationen). Für Gutartigkeit sprechen mittlere Echodichte des Tumorzentrums in Kombination mit indifferentem Schallverhalten (13/61 malignen, 33/48 benignen Lokalisationen) sowie regelmäßiger Gefäßdurchmesser bei diffuser Gefäßverteilung (3/36 malignen, 14/29 benignen Lokalisationen). Es konnte dargestellt werden, dass sich mit Hilfe der hochauflösenden B-Mode-Sonographie Kriterien aufzeigen lassen, die tendenziell für Gut- oder Bösartigkeit eines Mammatumors sprechen. Dabei ist es zweckmäßig, mehrere Parameter in die Beurteilung einfließen zu lassen. Auch die farbkodierte Duplexsonographie kann dabei einen Beitrag leisten. Die Ermittlung von Resistance- und Pulsatilitätsindex hingegen erweist sich als nicht sinnvoll. Ein Parameter, welcher in der Humanmedizin eine entscheidende Rolle zur Unterscheidung bösartiger von gutartigen Tumoren der Mamma spielt ist die Randkontur eines Tumors. Dies ist das einzige Kriterium, das auch bei Mammatumoren der Hündin einen diagnostischen Nutzen aufweist. Anhand einzelner sonographischer Parameter ist es nicht möglich, die Dignität eines Tumors vorherzusagen. Die sonographische Untersuchung kann jedoch in einigen Fällen beim Abschätzen der Prognose helfen.
In 53 bitches that underwent surgery because of tumors of the mammary gland at the Department of small animal medicine of the University of Leipzig we carried out a preoperative ultrasonographic examination of the mammary gland. Furthermore eight pregnant and one lactating bitch were examined the same way. We aimed to find out, whether high-resolution ultrasound helps differentiate benign from malignant tumors. Also we wanted to evaluate criteria established for that purpose in human medicine. Use of colour-coded duplex sonography, resistance index and pulsatility index for this question are reassessed too. The total number of mammary complexes examined for this study is 114. A GE Logiq™ 9 with a 14 MHz linear array transducer was used for all examinations. Seventy of the 114 sites of mammary tissue underwent a colour-coded duplex sonography after the B scan. Blood vessels were detectet in 70 of the tumors. In 47 of these sites the PW-Doppler was used to gain flow patterns to achieve resistance- and pulsatility-index. The images were analysed retrospectively. In B scan images lesions were judged by 12 parameters. Additional information about number, diameter and distribution of vessels within a tumor was taken from the images of colour-coded duplex sonography. The excised complexes were evaluated pathohistologically. Only descriptive statistical analysis was possible because the resulting groups were very small after being sorted according to WHO-classification. Therefore the complexes of mammary glands were subsumpted into two groups – „malignant“ and „benign“ tumours. An irregular contour of the tumor (32 of 61 malignant, 4 of 48 benign tumors), signal enhancement (36/61 malignant, 9/48 benign tumors) or total shadowing (8/61 malignant, 0/48 benign tumors) behind the tumor, calcification (20/61 malignant, 6/48 benign tumors) and irregular vessel diameters (25/61 malignant, 12/48 benign tumors) are signs of malignancy. Tumors that miss a clearly detactable borderline (15/61 malignant, 36/48 benign tumors) and tumors with no signal alteration behind the tumor (17/61 malignant, 39/48 benign tumors) are benign more often. The combination of parameters reduces the number of adequate tumors and rises significance. A tumor showing an irregular contour and calcification (13/61 malignant, 1/48 benign tumors) is more likely to be malignant as well as a tumor of medium echodensity showing signal enhancement (21/61 malignant, 6/48 benign tumors). Tumors of medium echodensity without signal alteration behind the lesion (13/61 malignant, 33/48 benign tumors) and tumors with diffusely distributed vessels of regular diameter (3/36 malignant, 14/29 benign tumors) are more likely to be benign. It could be shown that high-resolution B scan parameters can help differentiate between malignant and benign tumors of the mammary gland, especially if they are used in combination with each other. Parameters from colour-coded duplex sonography can increase predicting value of B scan examinations too but there is no use of analysing resistance index or pulsatility index. One of the criteria established in human medicine ist the contour of a tumor. This parameter is of diagnostic use in mammary tumours of the bitch too. It is not possible to clearly predict the character of a tumor of the mammary gland of a bitch by only a few parameters based on a sonogram but sonographic examination can be helpful for assessing prognosis sometimes.