Submitted by Ana Rosa Silva (arosa@ufpa.br) on 2012-07-23T13:47:00Z No. of bitstreams: 2 Dissertacao_ExpressaoGenesC-myc.pdf: 1582070 bytes, checksum: ded639d2c11fa3ed553f5b611b74750c (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Approved for entry into archive by Ana Rosa Silva(arosa@ufpa.br) on 2012-07-23T13:59:27Z (GMT) No. of bitstreams: 2 Dissertacao_ExpressaoGenesC-myc.pdf: 1582070 bytes, checksum: ded639d2c11fa3ed553f5b611b74750c (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Made available in DSpace on 2012-07-23T13:59:27Z (GMT). No. of bitstreams: 2 Dissertacao_ExpressaoGenesC-myc.pdf: 1582070 bytes, checksum: ded639d2c11fa3ed553f5b611b74750c (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Previous issue date: 2010 / O câncer de mama é um dos tumores de maior incidência na mulher, e por isso, muitas pesquisas vêm sendo realizadas, desde a avaliação das características epidemiológicas, à dinâmica biocelular e o tratamento desta doença. Na avaliação de respostas ao tratamento, os fatores preditivos são marcadores que auxiliam na escolha da melhor droga a ser usada. Esta dissertação teve o objetivo de avaliar os genes de receptores de estrogênio e progesterona, HER-2 e C-MYC em tumores localmente avançados da mama, como fatores preditivos de resposta à quimioterapia neoadjuvante. Estudaram-se fragmentos da neoplasia maligna mamária de 50 pacientes com carcinoma ductal infiltrativo, com estadiamento clínico E-III e tratadas com quimioterapia neoadjuvante. Utilizaram-se as técnicas de imunohistoquímica e de hibridização in situ por fluorescência (FISH). A análise dos receptores hormonais não apresentou diferença estatisticamente significativa comparando as pacientes com resposta satisfatória à quimioterapia, das insatisfatórias; a análise do HER-2 apresentou significância apenas para as respostas satisfatórias, onde houve baixa amplificação deste gene. Em relação ao C-MYC observou-se uma diferença estatisticamente significativa comparando a alta amplificação deste gene a uma resposta insatisfatória à quimioterapia. O estudo concluiu que o gene C-MYC pode ser um importante marcador de predição nos tratamentos quimioterápicos neoadjuvantes usados em câncer mamário. / Breast cancer is a higher incidence of tumors in women, and therefore many studies have been performed since the assessment of the epidemiological characteristics, the dynamics biocelular and treatment of this disease. In evaluating responses to treatment, the risk factors are markers that help in choosing the best drug to use. This work aimed to evaluate the genes of estrogen and progesterone receptors, HER-2 and C-MYC in locally advanced breast tumors as predictors of response to neoadjuvant chemotherapy. We studied fragments of mammary malignancy in 50 patients with infiltrating ductal carcinoma, with clinical stage III and E-treated with neoadjuvant chemotherapy. We used the techniques of immunohistochemistry and fluorescence in situ hybridization (FISH). The analysis of hormone receptors showed no statistically significant difference comparing patients with satisfactory response to chemotherapy, the poor and the analysis of HER-2 showed significance only for satisfactory answers, where there was poor amplification of this gene. Regarding C-MYC was observed a statistically significant difference comparing the high amplification of this gene to a poor response to chemotherapy. The study concluded that the C-MYC gene may be an important marker for predicting the treatments used in neoadjuvant chemotherapy for breast cancer.

Amplification of the receptor tyrosine kinase ErbB-2 has been linked to the proliferation of breast cancer cells.1,2 Trastuzumab targets the extracellular domain of ErbB-2, leading to growth inhibition of approximately 15% of the breast cancers with genomic amplification of the ERBB2 gene.3 Clinical studies have demonstrated its efficacy in both early4 and metastatic breast cancers. 5,6 However, many tumors with ERBB2 amplification are not responsive to treatment.7 Moreover, the ones that initially respond, eventually progress and acquire drug resistance.8 An in vitro model for this acquired resistance was established by Chan & al.9 The breast cancer cell line, BT474, containing amplified ERBB2, was grown in the presence of trastuzumab for several months until subclones outgrew. Gene expression profiling was performed on these clones to determine differentially expressed genes between the parental and resistant cells. DARPP-32 (Dopamine and cAMP regulated phosphoprotein of 32kDa) was, by far, the most overexpressed transcript. DARPP-32 is coamplified with ERBB2 on the same amplicon of chromosome 17.10 This protein has been mostly described in neurobiology, but DARPP-32 overexpression was recently reported in gastrointestinal, esophageal, prostate and breast cancer.11 Therefore, we suggest that overexpression of DARPP-32 can cause acquired resistance of breast cancer cells to trastuzumab. The in vitro knockout of DARPP-32, using stable shRNA transfection, abolishes the resistance to trastuzumab in the clones, while overexpression of DARPP-32 in the parental cells results in de novo resistance. Overall, our results suggest that DARPP-32 may be a potential therapeutic target in breast cancer patients who develop acquired trastuzumab resistance.

L'inhibidor de carboxipeptidasa de patata (PCI) és un antagonista de l'EGF. Un dels problemes del PCI, però, és la baixa afinitat per l'EGFR en comparació amb el lligand natural. En aquest treball s'han dissenyat nous bloquejadors de la via ErbB partint de l'estructura de l'EGF per tal d'augmentar l'afinitat d'unió. S'ha determinat que les variants tenen reduïda la capacitat d'activació del receptor però segueixen promovent la proliferació cel·lular. Tot i així, s'ha obtingut una variant, l'EGF Truncat, que tot i no estar correctament plegada, és la que ofereix uns millors. També s'ha dut a terme un tractament combinat entre el PCI i un inhibidor de tirosin quinases d'aquesta mateixa via, l'Iressa. Els resultats han posat de manifest que hi ha un antagonisme entre els dos tractaments. Aquesta és la primera vegada que s'associa de forma directa un tipus de tractament amb una resistència associada a l'activació de la producció autocrina de lligands, i representa un avenç molt important en la recerca oncològica. / Potato carboxypeptidase inhibitor (PCI) is an EGF antagonist. One of the main problems of PCI is its low affinity for EGFR in comparison to the natural ligand. In this study, blockers of the ErbB pathway have been designed taking as a model the EGF structure, in order to increase binding affinity. It has been observed that the variants have less ability to activate the receptor but they still promote cell proliferation. Nevertheless, the variant called Truncated EGF, although being unfolded, is the one that shows best perspectives.A combined treatment using PCI and a tyrosine kinase inhibitor of the same pathway, Iressa, has also been carried out. Results have shown that there is an antagonism between both treatments. This is the first time that a treatment is associated to resistance involving autocrine loop activation of ligands, and it represents a very important finding in the oncologic research.

Diss. (sammanfattning) Umeå : Umeå universitet, 2006. / Härtill 5 uppsatser.

INTRODUÇÃO: O câncer do pâncreas apresenta taxas anuais de mortalidade e incidência muito semelhantes, sendo uma das principais causas de morte por câncer no mundo. A agressividade do tumor e o retardo no seu diagnóstico são considerados responsáveis pela sua alta letalidade. O tratamento adjuvante convencional aumenta pouco a sobrevida a longo prazo e a terapia-alvo pode ser uma alternativa ao tratamento deste tipo de tumor. OBJETIVO: O objetivo do presente estudo é avaliar a expressão do receptor do fator de crescimento epitelial e seu eventual valor prognóstico em pacientes portadores de adenocarcinoma pancreático submetidos à ressecção cirúrgica. MÉTODO: Foram estudados retrospectivamente 88 pacientes portadores de adenocarcinoma pancreático operados no Serviço de Cirurgia de Vias Biliares e Pâncreas do HC-FMUSP e no Departamento de Cirurgia Abdominal Hospital A.C. Camargo no período de 1990 a 2006. RESULTADOS: Quarenta e sete (53,4%) pacientes do sexo feminino e 41 (46,6%) do masculino com idade mediana de 60 anos. As cirurgias realizadas foram duodenopancreatectomia com preservação do piloro (55,1%), gastroduodenopancreatectomia (34,8%), pancreatectomia corpo-caudal (6,8%) e gastroduodenopancreatectomia total (2,3%). A ressecção venosa portal foi realizada em 12 pacientes (13,5%). O tamanho tumoral médio foi de 3,75cm. Invasão vascular esteve presente em 31% dos casos e neural em 88,5%. A margem de ressecção estava comprometida em 33 pacientes (37,5%). Cinco (5,7%) pacientes eram do estádio IA, 15(17%) do IB, 19(21,6%) do IIA, 47(53,4) do IIB e dois (2,3%) do III.Observou-se diferença na expressão de EGFR na membrana celular entre o tecido tumoral e o tecido não tumoral (p=0,004); entre o tecido metastático linfonodal e o tecido não tumoral (p=0,02) mas não houve diferença quanto à sua expressão quando comparamos o tecido tumoral e o tecido metastático linfonodal (p=0,28). Dentre as variáveis clínicas e patológicas estudadas, observou-se diferença de expressão do EGFR entre os gêneros feminino e masculino (p=0,03), não havendo diferenças entre as outras variáveis. A sobrevida mediana global foi de 22,9 meses. A sobrevida cumulativa global em 1 ano, 3 anos e 5 anos foi de 48%, 20% e 18%, respectivamente. As sobrevidas cumulativas em 1 ano, 3 anos e 5 anos foram 77%, 30% e 8% no grupo sem expressão do EGFR na membrana tumoral versus 46%, 8% e 0% respectivamente no grupo com expressão do EGFR na membrana celular do tumor. Na análise univariada, as seguintes variáveis estiveram associadas a menor sobrevida: sexo masculino, ressecção venosa portal, invasão peri-neural, e vascular, invasão do tecido peri-pancreático, acometimento da margem de ressecção pancreática e expressão positiva de EGFR no tecido tumoral. Na análise multivariada, os fatores associados à sobrevida menor foram: gênero masculino, ressecção venosa portal, invasão vascular e invasão peri-neural. CONCLUSÃO: A expressão do EGFR na membrana celular é significativamente maior no tecido tumoral que no tecido pancreático não tumoral. A expressão do EGFR na membrana celular do tecido tumoral está associada a pior prognóstico (menor sobrevida). / INTRODUCTION: Pancreatic cancer is one of the main cancer related deaths in the world and its incidence is similar to its mortality. Biological aggressiveness and delayed diagnosis are a major concern. Adjuvant treatment has little impact on survival and the expression of potential target molecules has been undertaken in order to increase survival. OBJECTIVE: The aim of the present study is to study the expression of EGFR and its potential prognostic role in tumor, non-tumor and metastatic lymph node tissues of patient with pancreatic adenocarcinoma treated with surgical resection. MATHERIAL AND METHODS: Eighty eight patients with pancreatic adenocarcinoma operated at Serviço de Cirurgia de Vias Biliares e Pâncreas do HC-FMUSP and Departamento de Cirurgia Abdominal do Hospital A.C.Camargo were retrospectively studied between 1990 and 2003. RESULTS: Forty seven females (53,4%) and 41 males (46,6%) with median age of 60 years were studied. Pylorus preserving duodenopancreatectomy was performed in 55%, classical duodenopancreatectomy in 34,8%, distal pancreatectomy in 6,8% and total pancreatectomy in 2,3%. Portal vein resection was performed in 12 patients (13,5%). Mean tumor size was 3,75cm. Vascular and neural invasion were present in 31% and 88,5%, respectively. Positive surgical margin was present in 33 (37,5%) patients. Five (5,7%) patients were stage IA, 15(17%) stages IB, 19(21,6%) stages IIA, 47(53,4%) stages IIB and two (2,3%) stages III. There were difference in the membrane expression of EGFR between tumor and non tumor pancreatic tissue (p=0,004); between metastatic lymph node and non tumor pancreatic tissue (p=0,02); but there were no difference between tumor and metastatic lymph node tissue (p=0,28). Median survival time was 22,9 months. Cumulative one, three and five years survival were 48%, 20% and 18%. Cumulative survival at 1, 3 and 5 years were 77%, 30% and 8% in patients with negative expression of EGFR in tumor membrane and 46%, 8% and 0%, respectively in patients with positive EGFR expression in tumoral membrane. Univariate analysis showed that male gender, portal vein resection, neural, vascular and peri-pancreatic invasion invasion, positive surgical margin and positive membrane EGFR expression in tumoral tissue were correlated with poor survival. Multivariate analysis showed that male gender, portal vein resection, vascular invasion and peri-neural invasion are associated with lower survival after resection. CONCLUSION: EGFR membrane expression is different between tumor tissue and non tumor pancreatic tissue. EGFR membrane expression in tumoral tissue was associated with worst survival.

No description available.

No description available.

Introdução: Estimativas mostram que mais de dois terços das mulheres com câncer de mama possuem receptores hormonais positivos, e recebem terapia endócrina como tratamento, sendo tamoxifeno (TAM) o tratamento padrão (EBCTCG 2005; Davies et al., 2012). Porém, muitas pacientes se tornam resistentes com o passar do tempo. Estudos prévios mostraram que a expressão do receptor de estrogênio β (REβ) aumenta a resposta ao tratamento com TAM em células de câncer de mama, assim como a coexpressão de REα e REβ esta associada com maior ação proliferativa de TAM (Treeck et al., 2010; Sun et al., 2014). Também foi observada a existência de “cross-talk” entre os RE e a família do receptor do fator de crescimento epidérmico (HER) na resposta ao tratamento com TAM (Lindberg et al., 2011; Blows et al., 2010). Objetivo: Verificar a expressão do REβ, e suas interações com REα e receptores HER, durante o tratamento com TAM e em células resistentes ao TAM. Métodos: A expressão do REβ foi analisada em dois bancos de dados contendo informações de pacientes com câncer de mama. A expressão de RNAm dos RE, receptores HER e vias de sinalização PTEN, Akt e MAPK foram avaliadas após tratamento com TAM, em células resistentes ao TAM e em células silenciadas para os genes dos RE. Também foi avaliada a viabilidade celular após tratamento com TAM e nas células silenciadas para os genes dos RE. Resultados: Pacientes com câncer de mama apresentaram expressão reduzida do REβ, e os subtipos de câncer de mama REα positivos apresentaram baixa expressão do REβ quando comparados aos subtipos REα negativos. Células expressando níveis moderados de REβ apresentaram melhor resposta ao tratamento com TAM. Diminuição nos níveis dos RE é acompanhada por aumento nos níveis dos receptores ErbB2 e ErbB3, aumento de PTEN e diminuição de Akt e MAPK3 após tratamento com TAM. ERβ modula a ação antiproliferativa do TAM através da via de MAPK3. Células resistentes ao TAM apresentaram baixos níveis dos RE e altos níveis dos receptores EGFR, ErbB3 e ErbB4. Conclusão: Estes resultados demonstram que o REβ, e suas interações com REα e receptores HER, possuem papel importante na resposta ao tratamento com TAM. / Introduction: Approximately two-thirds of all breast cancer patients overexpress hormonal receptors, and are treated with endocrine therapy, being tamoxifen (TAM) the standard treatment. However many of initial responders to TAM as first-line experience relapse. Several mechanisms have been proposed to explain the occurrence of acquired TAM resistance. Previous studies showed that estrogen receptor β (ERβ) expression is associated with better response to tamoxifen treatment, as the co-expression of ERα and ERβ is associated with TAM antiproliferative effects. Moreover, there is growing interest about the cross-talk between ERs and ErbB family in response to endocrine therapy. Suggesting that TAM can acts through ERβ and/or ErbB family as compensatory pathways. Objective: To evaluate the expression of ERβ and the relation of ERβ with ERα and ErbB family in response to TAM treatment and in TAM resistant cells. Methods: ERβ expression was analyzed in two different databases of breast cancer patients. The mRNA levels of ER, HER receptors and PTEN, Akt and MAPK signal pathways were measured after TAM treatment, in TAM resistance cells and in cells silenced for ER genes. The cellular viability was also measured after TAM treatment, in TAM resistance cells and in cells silenced for ER genes. Results: Breast cancer patients presented reduced ERβ expression and the ERα-positive breast cancer subtypes presented lower ERβ levels when compared to ERα-negative breast cancer subtypes. Cells expressing moderates levels of ERβ presented better response to TAM treatment. Down-regulation of ERs induced by TAM treatment are accompanied with an increase in ErbB2 and ErbB3, reduced AKT and MAPK3 mRNA levels and increased PTEN levels. ERβ modulates TAM anti-proliferative effects through MAPK3 pathway. TAM– resistant cells expressed decreased ER mRNA levels and increased EGFR, ErbB3 and ErbB4 levels. Demonstrating that the cross-talk between ERs and HER family influence the response to TAM treatment. Conclusion: These results provide additional data indicating the importance of ERβ, and the relation with ERα and HER receptors, to predict TAM responsiveness.

Abstract Cancer treatments have remarkably improved over the past years since targeted therapies and immunotherapy have been introduced to the field of oncology. The benefit of these new therapies is often limited, however, by de novo or acquired therapy resistances, which should be noticed when making clinical decisions. In this current work, we studied the prognostic and predictive values of several immunological markers in metastatic HER2-positive breast cancer treated with trastuzumab, because trastuzumab is still given to patients according to the HER2 status only, without certainty of tumor response. We also determined the role of HER2 and HER3 for cancer stem cells (CSC) in ALK translocated non-small cell lung cancer (NSCLC) cell lines since the CSCs are causing therapy resistance and cancer recurrence. The results demonstrated that a high number of cytotoxic T cells, together with a high number of M1-like macrophages in the center of the tumor (CT), are promising and independent prognostic factors in HER2-positive breast cancer. These markers together also can predict the progression of the disease and the length of trastuzumab discontinuation in tumor response. Expression of HER2 and HER3 increased the stem-like properties of ALK translocated NSCLC cells, which were decreased when the expressions were downregulated. HER2-HER3-dependent CSCs also mediated the ALK therapy resistance. In conclusion, this study suggests that patients with a favorable immunological tumor profile (high number of cytotoxic T cells and M1-like macrophages in the CT) could be treated in a less-intensive manner, that trastuzumab discontinuation could be feasible for these patients, and that targeting of HER2 and HER3 receptors can lead to more effective killing of cancer stem-like cells and should be further studied. / Tiivistelmä Syöpähoidot ovat kehittyneet huomattavasti, kun kohdennetut hoidot ja immunologiset hoidot ovat tulleet perinteisten hoitojen rinnalle. Usein näiden hoitojen hyötyä kuitenkin rajoittaa jo olemassa oleva lääkeresistenssi tai sen kehittyminen, mikä tulisi ottaa huomioon hoitoja suunniteltaessa. Tässä työssä tutkittiin immunologisia merkkiaineita, joilla voitaisiin ennustaa trastutsumabi-hoidon vastetta sekä potilaiden ennustetta levinneessä HER2-positiivisessa rintasyövässä. Tällä hetkellä trastutsumabi-hoitopäätös tehdään pelkän HER2-geenimonistuman mukaan ilman varmuutta siitä, hyötyykö potilas oikeasti hoidosta. Lisäksi tutkimme HER2- ja HER3-reseptorien merkitystä syövän kantasoluille ALK-translokoituneessa ei-pienisoluisessa keuhkosyövässä (NSCLC), sillä syövän kantasolut ovat yksi merkittävimmistä tekijöistä lääkeresistenssin kehittymisessä ja syövän uusiutumisessa. Työssä havaittiin, että kasvaimen keskellä oleva suuri määrä sytotoksisia T-soluja sekä M1-tyypin makrofageja on yhteydessä potilaiden parempaan ennusteeseen ja että kyseiset merkkiaineet ovat toisistaan riippumattomia. Merkkiaineet pystyivät ennustamaan myös taudin etenemistä sekä trastutsumabi-hoitokeskeytyksen pituutta. HER2- ja HER3-proteiinien tuotto lisäsi ALK-translokoituneiden NSCLC-solujen kantasolumaisia ominaisuuksia, jotka puolestaan vähenivät, kun proteiinien tuotto estettiin. Lisäksi HER2-HER3 -riippuvaiset syövän kantasolut säätelivät lääkeresistenssiä kyseisessä taudissa. Työn tulokset viittaavat siihen, että potilaita, joilla on suotuisa kasvaimen immunoprofiili (suuri määrä sytotoksisia T-soluja ja M1-tyypin makrofageja kasvaimen keskellä) pystyttäisiin hoitamaan keveimmillä hoidoilla ja HER2-hoitokeskeytys voisi olla mahdollinen näillä potilailla. Lisäksi työ korostaa HER2- ja HER3-reseptorien kohdentamista syövän kantasolumaisten solujen tehokkaamman tuhoamisen saavuttamiseksi. / Huomautus/Notice Painetussa virheelliset ISBN -tunnukset: ISBN (print) 978-952-42-2343-8 pitäisi olla 978-952-62-2343-8. ISBN (PDF) 978-952-42-2344-5 pitäisi olla 978-952-62-2344-5. Printed version has incorrect ISBNs: ISBN (print) 978-952-42-2343-8 it should be 978-952-62-2343-8. ISBN (PDF) 978-952-42-2344-5 it should be 978-952-62-2344-5.

<p>The incidence of breast cancer is rising steadily in Sweden and the proportion of carcinoma in situ (CIS) has increased appreciably, most likely due to mammography screening. The aim of this study was twofold: (1) to examine risk factors for subsequent invasive breast carcinoma and breast cancer death after primary ductal carcinoma in situ (DCIS) and (2) to study the biology in the progress between in situ and invasive carcinoma.</p><p> In a cohort-study based on 3,398 women with a primary CIS reported to the Swedish Cancer Registry (SCR) 1980-1992, women diagnosed in 1989-1992 ran a relative risk of 0.1 (CI 95%, 0.0-0.9) from dying of breast cancer as compared with women diagnosed in 1980-1982. Women in counties with mammography screening ran a relative risk of 0.2 (CI 95%, 0.0-2.1) for breast cancer death in comparison with women in non-screening counties.</p><p> In a case-control study derived from all 4,661 women with primary CIS reported to the SCR 1960-1992, we investigated risk factors for subsequent invasive breast carcinoma (n=118) and breast cancer death (n=39). Large size and multifocality were found to increase the risk for breast cancer death. Postoperative radiotherapy and mastectomy lowered the risk for ipsilateral invasive cancer.</p><p> The standardised incidence rates (SIR) for invasive breast cancer were estimated in the cohort from 1980-1992. The SIR after primary DCIS and primary lobular carcinoma in situ (LCIS) was 4.5 (CI 95%, 3.7-5.5) and 4.0 (CI 95%, 2.1-7.5), respectively.</p><p> New histopathological classification systems for DCIS were evaluated in 195 women consecutively diagnosed with primary DCIS between 1986-1994. One group with highly differentiated lesions was defined with the EORTC classification system and had an excellent prognosis.</p><p> Histopathological grade and expression of p53, c-erbB-2, Ki 67, hormone receptors, Bcl-2 and angiogenesis were compared in 626 women with either a pure DCIS, a small invasive carcinoma or a lesion with both an invasive and in situ component. When grade was taken into account, no change in tumour markers could be detected that signalled the progression from an in situ stage to invasiveness. All tumour markers correlated to grade and their distribution was very similar in the two components of mixed lesions.</p>