Immunological Characteristics between αß TDC and γδ TDC Cells in the Spleen of Breast Cancer-Induced Mice / Características imunológicas entre células TDC αß e TDC γδ no baço de camundongos com câncer de mama induzido

Abstract Objective To evaluate the antitumoral role of γδ TDC cells and αβ TDC cells in an experimental model of breast cancer. Methods Thirty female Balb/c mice were divided into 2 groups: control group (n=15) and induced-4T1 group (n=15), in which the mice received 2 x 105 4T1 mammary tumor cell line. Following the 28-day experimental period, immune cells were collected from the spleen and analyzed by flow cytometry for comparison of αβ TDC (TCRαβ+ CD11c+MHCII+) and γδ TDC (TCRγδ+CD11c+MHCII+) cells regarding surface markers (CD4+ and C8+) and cytokines (IFN-γ, TNF-α, IL-12 and IL-17). Results A total of 26.53% of γδ TDC- control group (p<0.0001) - the proportion of αβ TDC was lower in splenic cells than γδ TDC; however, these 2 cell types were reduced in tumor conditions (p<0.0001), and the proportion of IFN-γ, TNF-α, IL-12 and IL-17 cytokines produced by γδ TDC was higher than those produced by αβ TDC, but it decreased under conditions of tumor-related immune system response (p<0.0001). Conclusion Healthy mice engrafted with malignant cells 4T1 breast tumor presented TDC with γδ TCR repertoire. These cells express cytotoxic molecules of lymphocytes T, producing anti-tumor proinflammatory cytokines.

Resumo Objetivo Esclarecer o possível papel antitumoral das células TDC γδ e TDC αβ em um modelo experimental de câncer de mama. Métodos Trinta baços de camundongos Balb/c analisados por citometria de fluxo, separados entre grupo controle (n=15) e o grupo tumoral induzido por 4T1 (n=15). Resultados Presença de 26,53% de TDC γδ nos camundongos do grupo controle (p<0,0001), proporção de TDC αβ menor em células esplênicas do que TDC γδ; no entanto, estes dois tipos de células são reduzidos emcondições tumorais (p<0,0001), e a proporção de citocinas IFN-γ, TNF-α, IL-12 e IL-17 produzidas pelas célula TDC γδ foi maior do que as produzidas pelas células TDC αβ, mas foram diminuídas sob condições de resposta ao sistema imunológico relacionada ao tumor (p<0,0001). Conclusão Camundongos saudáveis induzidos ao tumor de mama 4T1 apresentaram TDC com repertório TCR γδ. Estas células expressam moléculas citotóxicas de linfócitos T, produzindo citocinas proinflamatórias anti-tumor.

Análise de marcadores sanguíneos relacionados a prognóstico em câncer de reto localmente avançado / Analysis of prognostic-related blood markers in locally advanced rectal cancer

Câncer colorretal (CCR) é altamente incidente; estimam-se, no Brasil, em 2016 16.660 casos novos de CCR em homens e 17.620 em mulheres, de acordo com o Ministério da Saúde. Estudos têm demonstrado que as células tumorais circulantes (CTCs) podem estar envolvidas no processo de metastatização. Desta forma, seu isolamento constitui uma estratégia potencial para o acompanhamento clínico, como método não invasivo. O presente estudo teve como objetivos: 1) Isolar e quantificar CTCs do sangue periférico de pacientes com câncer de reto localmente avançado (CRLA), estádios II e III, para correlacionar seus níveis com exames de imagem e sobrevida livre de doença; 2) Analisar nas CTCs marcadores de resistência ao tratamento (TYMS e RAD23) e correlacionar com resposta à terapia e sobrevida livre de doença. Foram analisadas amostras de 30 pacientes. As amostras foram coletadas antes da quimioterapia neoadjuvante (baseline) e após a mesma (follow-up). As CTCs foram caracterizadas nos dois momentos por imunocitoquímica com anticorpos específicos para os marcadores acima citados e por CISH, avaliando a expressão do mRNA do gene do TYMS. As contagens de CTCs diminuíram entre a primeira e a segunda coletas em pacientes exibindo resposta patológica completa (RPC; p = 0,02) ou resposta parcial (RP; p = 0,01). Em relação à expressão protéica, TYMS estava ausente em 100% das CTCs dos pacientes com RPC (p = 0,001), mas foi expresso em 83% dos não respondedores na segunda coleta (p <0,001). Em paralelo, RAD23b foi expresso em CTCs de 75% dos não respondedores na coleta baseline (p = 0,01) e em 100% dos não respondedores do follow-up (p = 0,001). Surpreendentemente, 100% dos não respondedores expressaram mRNA de TYMS nos dois momentos (p = 0,001). Além disso, TYMS / RAD23b não foram detectadas nas CTCs de pacientes que exibiam RPC (p = 0,001). Encontramos 83,3% de sensibilidade para o mRNA de TYMS no baseline (p = 0,001) e 100% para a expressão da proteína TYMS (p = 0,064) e RAD23B (p = 0,01) no follow- up. Assim, a expressão do RNAm TYMS e / ou TYMS / RAD23b nas CTCs, bem como a cinética das CTCs, têm o potencial de prever não resposta ao tratamento neoadjuvante e evitar cirurgias radicais desnecessárias em pacientes com CRLA com RPC. Com a realização deste trabalho obtivemos uma melhor compreensão a respeito dos mecanismos que envolvem a resistência ao tratamento nos pacientes com CRLA, por meio da análise de CTCs. Auxiliados por estas análises, identificamos novos biomarcadores sanguíneos prognósticos, que poderão ser usados como instrumentos de direcionamento clínico na escolha da melhor terapêutica

Colorectal cancer (CRC) is highly incident; In Brazil, in 2016, an estimated 16,660 new cases of CRC in men and 17,620 in women, according to the Ministry of Health. Studies have shown that circulating tumor cells (CTCs) may be involved in the metastasization process. Thus, its isolation is a potential strategy for clinical follow-up as a noninvasive method. The present study aims to: 1) Isolate and quantify peripheral blood CTCs from patients with locally advanced rectal cancer (stages II and III) to correlate their levels with imaging and progression-free survival; 2) Analyze treatment resistance markers on CTCs (TYMS and RAD23), and correlate with therapy response and progression-free survival. Samples from 30 patients were analyzed. Samples were collected before and after neoadjuvant chemotherapy (follow-up). CTCs were characterized at both times by immunocytochemistry with specific antibodies to the markers mentioned above and by CISH, evaluating the expression of the TYMS gene mRNA. CTC counts decreased between the first and second collection in patients exhibiting complete pathological response (CPR; p = 0.02) or partial response (PR; p = 0.01). Regarding protein expression, TYMS was absent in 100% of the CTCs of patients with CPR (p = 0.001), but was expressed in 83% of non-responders in the second collection (p <0.001). Meanwhile, RAD23b was expressed in CTCs of 75% of baseline non-responders (p = 0.01) and 100% of follow-up non-responders (p = 0.001). Surprisingly 100% of non-responders expressed TYMS mRNA at both times (p= 0.001). In addition, TYMS / RAD23b was not detected in CTCs of patients with CPR (p = 0.001). We found 83.3% sensitivity for TYMS mRNA at baseline (p = 0.001) and 100% for TYMS protein expression (p = 0.064) and RAD23B (p = 0.01) at follow-up. Thus, TYMS mRNA and/or TYMS / RAD23b in CTCs, as well as CTC kinetics, have the potential to predict non-response to NCRT and to avoid unnecessary radical surgery in patients with LARC and CPR. With this work we obtained a better understanding about the mechanisms involving resistance to treatment in patients with locally advanced rectal cancer, through the analysis of CTCs. With these analyzes, we identified new prognostic blood biomarkers that could be used as clinical guidance tools in choosing the best therapy

Antiprogestágenos en cáncer de mama: del ratón a la clínica y nuevamente al ratón / ANTIPROGESTINS IN BREAST CANCER : FROM MOUSE TO HUMANS AND BAC

En el laboratorio de la Dra Pasqualini, por serendipitismo surgió un modelo de cáncer de mama murino que se caracteriza por el hecho de que es uno de los pocos modelos de ratón en los cuales los carcinomas mamarios inducidos modelan al tipo de tumor más frecuente de la mujer: el carcinoma mamario luminal que expresa receptores hormonales y es modulable por terapia endócrina. Luego ya en el IBYME demostramos que en este modelo los receptores de estrógeno interaccionan con los de progesterona en los promotores de genes blanco y que la proporción de isoformas de receptor de progesterona es fundamental para predecir la respuesta a antiprogestágenos. Hemos validado estas observaciones en modelos de xenotransplantes de líneas celulares de cáncer de mama humano creciendo en ratones inmunosuprimidos y posteriormente, utilizando muestras de pacientes de cáncer de mama humano, hemos demostrado que sólo aquellas con mayor expresión de isofoma A que de isofoma B, responden ex vivo al tratamiento con antiprogestágenos. Proponemos que estas pacientes serían candidatas a tratamientos combinados de antiestrógenos y antiprogestágenos. Estudios de tumores de pacientes creciendo en ratones inmunosuprimidos ayudarán a discernir cuáles tumores responden mejor a una u otra terapia endocrina

By serendipity, at Dr. Christiane Dosne Pasqualini´s Laboratory,, we have developed one of the few mouse experimental models of breast cancer in which induced-tumors share many features of human luminal breast cancer: ductal histology, hormone receptor expression and hormone responsiveness. After moving to IBYME, we were able to demonstrate that estrogen receptors interact with progesterone receptors at the promoter of key target genes and that the ratio of progesterone receptor isoforms A and B is essential to predict antiprogestin responsiveness. After validating this data in human breast cancer cell xenografts, modified to express different PR isoform ratios, we designed a study aimed to evaluate the response of human breast cancers to antiprogestins ex-vivo and correlate this data with the PR isoform ratio. Only those samples with higher levels of isoform A than isoform B were inhibited by antiprogestins. We propose that selected breast cancer patients, those with higher levels of isoform A than isoform B, might be candidates for adjuvant treatment with antiprogestins in combination with standard endocrine therapy. Future experiments generating patient derived xenografts will help to understand and better select patients for personalized endocrine treatments.

Protagonismo del sistema inmune en el microambiente de los tumores malignos de la mama / Role of the immune system in the microenvironment of malignant tumors of breast

Los avances recientes en la comprensión de los mecanismos génicos y moleculares del cáncer de mama han revelado que el sistema inmune protagoniza los eventos responsables del desarrollo y la progresión del tumor. Las células de la respuesta inmune innata y adaptativa, así como diversos mediadores solubles liberados por ellas, pueden establecer una respuesta antitumoral protectora o, por el contrario, inducir eventos de inflamación crónica que favorezcan la promoción y progresión de esta enfermedad. Esta dualidad, se protagoniza en el microambiente del tumor, el cual puede regular la carcinogénesis en dependencia del infiltrado de células inmunes que predominen. Esta revisión, pretende resumir los conocimientos actuales de la relación sistema inmune-cáncer de mama, enfatizando en las células inmunes del microambiente del tumor y su importancia como biomarcadores de evolución clínica de la enfermedad(AU)

The recent advances in the understanding of the genetic and molecular mechanisms of breast cancer have demonstrated that immune system plays important events responsible for the development and progression of the tumor. The cells of the innate and adaptive immune system, as well as diverse soluble mediators, may establish a protective anti-tumor response or, on the contrary, to induce events of chronic inflammation that favor promotion and progression of disease. This duality occurs in the tumor microenvironment, which can regulate the carcinogenesis depending on the predominant immune cells. This revision summarizes the current knowledge of the relationship between immune system - breast cancer, emphasizing in the immune cells of the tumor microenvironment and its importance as biological markers of the clinical evolution of the disease(AU)

Análisis de las variables clinicopatológicas e inmunohistoquímicas del cáncer de mama / Analysis of the clinicopathological and inmunohistochemical variables of breast cancer

Los carcinomas de mama representan un grupo heterogéneo de tumores tanto en comportamiento clínico como en pronóstico, estableciéndose su diversidad a nivel molecular al expresar distintos genes que le confieren variabilidad biológica.Objetivos: Clasificar los carcinomas de mama en subtipos moleculares mediante marcadores inmunohistoquímicos y analizar las características clinicopatológicas.Material y método: Se analizaron retrospectivamente 303 pacientes con diagnóstico de cáncer de mama invasivo derivadas a la Unidad Integral De Oncología Gral. Roca (Río Negro), entre los años 2008-2013. Las pacientes se clasificaron en cuatro subtipos tumorales: Luminal A, Luminal B HER2 Positivo, Triple Negativo y HER2. Resultados: La distribución por inmunofenotipos fue: Luminal A: 72,93%, Luminal B HER2 Positivo: 9,57%, Triple Negativo: 12,22% y HER2: 5,28 %. Los carcinomas Luminal A expresaron tumores más pequeños, diferenciados, con ganglios axilares negativos y estadio precoz de la enfermedad respecto de los subtipos Luminal B HER2 Positivo, Triple Negativo y HER2. Conclusiones: La clasificación del cáncer de mama basada en parámetros inmunohistoquímicos permite una mejor definición pronóstica. Los tumores Luminal A presentaron características clinicopatológicas más favorables que los inmunofenotipos Luminales B HER2 Positivos, Triple Negativo y HER2...

Caracterização do perfil de dano oxidativo em carcinoma mamário e avaliação de associação com parâmetroshistopatológicos / Profile characterization of oxidative damage in breast carcinoma and evaluation of association with histopathological parameters

O carcinoma mamário é uma doença complexa que engloba diversas alterações genéticas e bioquímicas, além de mudanças no microambiente tumoral, tais como o estresse oxidativo,que pode ocasionar danos estruturais em lipídios, proteínas, bem como no DNA nuclear. O presente estudo teve por objetivo caracterizar o perfil oxidativo de neoplasias mamárias,comparando o perfil de expressão de produtos dos processos de peroxidação lipídica, nitração proteica e oxidação de DNA em função de parâmetros histopatológicos com valor prognóstico quanto à evolução clínica do câncer de mama. (...) Contudo, quando o tamanho tumoral foi analisado separadamente entre tumores Triplo-Negativos ou entre os demais subtipos (Luminal A/B eHER2-like), não se observou efeito significativo sobre a expressão de 8-OHdG. Em conclusão, os dados indicam que alguns tumores Triplo-Negativos são mais resistentes a danos oxidativos sobre DNA, independentemente de grau ou tamanho tumoral, o que pode contribuir para o perfil de maior agressividade deste subtipo tumoral...

Breast cancer is a complex disease that involves multiple genetic and biochemical alterationsand changes in the tumor microenvironment, such as oxidative stress, which can lead tostructural damages to lipids, proteins and nuclear DNA. The present work aimed tocharacterize the oxidative profile of mammary tumors, comparing the products of profileexpression of lipid peroxidation processes, protein nitration and DNA oxidation according tohistopathological parameters with prognostic value as to clinical evolution of breast cancer. (...) However, when the tumor size was analyzed separately from Triple-Negativetumors or among the other subtypes (luminal A / B and HER2-like), there was no significanteffect on the expression of 8-OHdG. In conclusion, the data indicate that some triple-negativetumors are more resistant to oxidative damage over the DNA, regardless of tumor grade orsize, which may contribute to the more aggressive subtype of the tumor...

Effect of Propofol and Desflurane on Immune Cell Populations in Breast Cancer Patients: A Randomized Trial

Several factors can affect the perioperative immune function. We evaluated the effect of propofol and desflurane anesthesia on the surgery-induced immune perturbation in patients undergoing breast cancer surgery. The patients were randomly assigned to receive propofol (n = 20) or desflurane (n = 20) anesthesia. The total and differential white blood cell counts were determined with lymphocyte subpopulations before and 1 hr after anesthesia induction and at 24 hr postoperatively. Plasma concentrations of interleukin (IL)-2 and IL-4 were also measured. Both propofol and desflurane anesthesia preserved the IL-2/IL-4 and CD4+/CD8+ T cell ratio. Leukocytes were lower in the propofol group than in the desflurane group at 1 hr after induction (median [quartiles], 4.98 [3.87-6.31] vs. 5.84 [5.18-7.94] 10(3)/microL) and 24 hr postoperatively (6.92 [5.54-6.86] vs. 7.62 [6.22-9.21] 10(3)/microL). NK cells significantly decreased 1 hr after induction in the propofol group (0.41 [0.34-0.53] to 0.25 [0.21-0.33] 10(3)/microL), but not in the desflurane group (0.33 [0.29-0.48] to 0.38 [0.30-0.56] 10(3)/microL). Our findings indicate that both propofol and desflurane anesthesia for breast cancer surgery induce a favorable immune response in terms of preservation of IL-2/IL-4 and CD4+/CD8+ T cell ratio in the perioperative period. With respect to leukocytes and NK cells, desflurane anesthesia is associated with less adverse immune responses than propofol anesthesia during surgery for breast cancer. (Clinical trial registration at https://cris.nih.go.kr/cris number: KCT0000939)

Recurrencia luego del tratamiento conservador en cáncer de mama: su relación con la edad y factores inmunohistoquímicos / Recurrence after breast cancer conservative treatment: relationship with age and immunohistochemical factors

La estrategia terapéutica para los tumores de mama en estadios tempranos ha sido establecida desde fines del siglo XX; es entonces que al tratamiento quirúrgico conservador y la radioterapia se sumó el ganglio centinela y el tratamiento adyuvante, de acuerdo a parámetros pre- y posoperatorios. Sin embargo, existe en todas las series una tasa de recurrencia local, y se ha podido determinar en parte cuáles son los factores de la paciente, anatómicos e histológicos, que predisponen a tal evento, a partir de hallazgos posoperatorios.También existe una tasa de enfermedad regional y a distancia, derivando en la necesidad de identificar factores de predicción que puedan sugerir la necesidad de una terapéutica más agresiva en pacientes susceptibles. Por otra parte, la clasificación molecular de los tumores de mama descripta por Perou y cols. pone en evidencia características inherentes a cada tumor, permitiendo establecer subtipos con implicancia clínica. A partir del análisis de nuestra estadística, en la que presentamos pacientes con cáncer de mama inicial que han recibido un tratamiento conservador, los objetivos planteados son: Presentar las tasas de eventos locales, regionales, a distancia y sobrevida luego de 15 años de experiencia en tratamiento conservador, y determinar si existe en nuestra serie una relación entre estos eventos, la edad de las pacientes y la clasificación molecular de los tumores. Establecer a partir de los resultados su implicancia a nivel asistencial.

Presence of CD3+ tumor infiltrating lymphocytes is significantly associated with good prognosis in infiltrating ductal carcinoma of breast.

BACKGROUND: Aim of this study was to investigate the prognostic significance of CD3+ TILs in infiltrating ductal carcinoma (IDC) of the breast. MATERIALS AND METHODS: Immuno-histochemistry was done with CD3 antibodies in tissue sections of 127 breast cancer patients, and CD3+ intra-tumoral and stromal TILs were counted in relation to clinico-pathological variables. RESULTS: Intra-tumoral and stromal CD3+ TILs were significantly associated with positive lymph node status (P = 0.006, P = 0.043, respectively) without significant association with age, menopausal status, family history, and hormonal status. The higher CD3 intra-tumoral and stromal counts both showed significant association with good prognosis (P = 0.039, P = 0.044, respectively). The intra-tumoral count was higher than stromal count and was independently associated with disease-free survival in stage I and II cancer (P = 0.021). CONCLUSIONS: CD3+ TILs may serve as independent marker of good prognosis in IDC breast. The findings of this study need further validation on a larger sample size.

Sclareol reduces CD4+ CD25+ FoxP3+ t-reg cells in a breast cancer model in vivo

Sclareol is a phytochemical used in people's diet in Southeast Asia. To investigate the immunotherapeutic effectiveness of Sclareol against breast cancer by direct intraperitoneal injection. Sclareol was isolated and purified from Salvia sclarea. Effect of Sclareol on cell growth inhibition was evaluated by MTT assay. Intraperitoneally injected Sclareol effects on reducing the tumor volume and shifting the cytokine profile were investigated. We also assessed if intraperitoneally injected Sclareol could improve the outcome of cancer therapy through suppressing the regulatory T cells. The results confirmed a significant decrease in the tumor size. Furthermore, a significant decrease in the level of IL-4 and an increase in the level of IFN-gamma were noticed in the intraperitoneally injected Sclareol group [p<0.05]. It was also observed that the splenocytes of treated animals significantly increase in cell proliferation assay. Moreover, measurements of splenic T regulatory cell indicated that intraperitoneally injected Sclareol significantly decreased the number of splenic T regulatory cell. Our results suggest that Sclareol, by reducing T-reg cells frequency and also tumor size can enhance the effect of cancer therapy as an immunostimulant

Concomitant increase of OX40 and FOXP3 transcripts in peripheral blood of patients with breast cancer

Regulatory T cells [T-regs] have an important role in cancer by suppression of protective antitumor immune responses. Regulatory T cells express the forkhead/ winged helix transcription factor [FOXP3] and OX40 molecules which have important regulatory roles in the immune system. To evaluate FOXP3 and OX40 transcripts in the peripheral blood mononuclear cells of women with breast cancer. Blood samples from 40 women with histologically-confirmed infiltrating ductal carcinoma of the breast and 40 healthy volunteer women without a history of malignancy or autoimmune disorders were collected. The abundance of FOXP3 and OX40 gene transcripts were determined by quantitative real-time PCR [qRT-PCR]. There was a significant positive correlation between FOXP3 and OX40 gene expression in women with breast cancer in a stage dependent manner. This finding emphasizes the importance of T-regs as predominant targets for breast cancer immunotherapy

Immunohis1ochemical cyclooxygenase-2 [COX-2] and cd31 expression in breast carcinoma with correlation to clinico-pathological parameters

Breast cancer is the most'common cancer in Egyptian women. COX-2 seems to be involved in malignant transformation and tumor progression by affecting cell proliferation, mitosis, cell adhesion, apoptosis, immune surveillance, and angiogenesis. Angiogenesis is an important key step in tumor progression. Microvascular density [MVD], a surrogate marker of angiogenesis can be assessed by CD31 staining. This study aims to: 1. Evaluate COX-2 and CD31 expressions in breast cancer. 2. Determine the correlation between COX-2 and CD31 with the clinico-pathological parameters in ductal breast carcinoma. This study included 74 specimens of breast lesions. Patient's age, tumor size and local aggressive changes, history of recurrence and/or presence of distant metastasis were obtained. Hematoxylin and Eosin [HandE] stained sections were evaluated for histopathological tumor type, tumor grade, presence or absence of normal hyperplastic, in situ component, lymphocytic infiltration, lymphovascular invasion, and axillary lymph node status. COX-2 and CD31 immunostaining was done to detect their expression using the avidin-biotin peroxidase method. COX-2 increased with increasing grade of ductal carcinoma in situ [DC1S] and invasive ductal carcinomas [IDC] [P< 0.05 and P< 0.002 respectively]. COX-2 expression increased progressively along the continuum of neoplastic changes from normal breast epithelium to IDC [P< 0.01]. There was significant correlation between COX-2 and tumor size [P< 0.05], tumor grade [P< 0.002], lymphovascular invasion [P< 0.03] and lymph node metastasis [P< 0.02]. CD31 staining was observed along the cell membrane of endothelial cells of microvessels in all breast specimens. The median CD31 MVD count was 10 for normal breast, increased insignificantly to 17 in hyperplastic lesions, and reached 19 for DCIS, and 66.5 in IDC [P < 0.000]. There was significant increase in MVD between different grades of IDC [P < 0.01] but not in DCIS. Positive correlation was present between COX-2 and CD31 in DCIS and in IDC [P< 0.000 for each]. COX-2 was increased with poor prognostic parameters; tumor size, tumor grade, lymphovascular invasion and lymph node metastasis. CD31 increases with increasing grade of IDC. These findings might imply for new therapeutic strategies in order to prevent progression of DCIS to IDC and to improve cancer therapy

Estudo clínico, histopatológico e imunoistoquímico de neoplasias mamárias em cadelas / Clinical, histopathological and immunohistochemical study of mammary neoplasm in bitches

O objetivo foi realizar um estudo clínico e epidemiológico de neoplasias mamárias em cadelas, considerando-se histórico reprodutivo, exame físico, diagnóstico histopatológico e imunoistoquímico. Utilizaram-se 60 neoplasias mamárias, divididas em grupos (grupo 1 - benigno, e grupo 2 - maligno). Avaliaram-se dados do histórico reprodutivo, o exame físico e achados histopatológicos e imunoistoquímicos para fator de crescimento endotelial vascular. Ao estudo do histórico reprodutivo, encontraram-se 90% dos animais com irregularidade de cio, 86,63% das cadelas não foram medicadas com contraceptivos e 83,33% não eram castradas. Ao exame físico, não foi verificada diferença (p>0,05) entre grupos ao se avaliar consistência das massas, regularidade da superfície tumoral e localização anatômica dos tumores. Quanto ao tamanho das massas, verificou-se diferença entre os grupos (p=0,0077), com 0,78±1,13cm para o grupo 1 e 1,81±2,29cm para o grupo 2. Diagnosticaram-se 40% de massas benignas e 60% de malignas, de acordo com os tipos de neoplasias. Para VEGF, verificaram-se valores médios de 2,22±0,89 para tumores malignos e 1,66±0,91 para benignos, com diferença entre grupos (p=0,0315). As neoplasias mamárias em cadelas não apresentam características de histórico reprodutivo e de exame clínico que auxiliem o diagnóstico diferencial, sendo a histopatologia o único método para conclusão do diagnóstico e a imunoistoquímica podendo ser utilizada para prognóstico da lesão.

The objective was to conduct a clinical and epidemiological study of mammary cancer in bitches, considering their reproductive history, physical examination, histopathological and immunohistochemical diagnosis. We used 60 breast tumors which were divided into groups (group 1 - group 2 and benign - malignant). We evaluated data from the reproductive history, physical examination and histopathology and immunohistochemistry for VEGF. The study of reproductive history had 90% of irregular estrus, 86.63% of the dogs were not tested with contraceptives and 83.33% were not castrated. On physical examination, there was no difference (p>0.05) between groups regarding the consistency of the masses, surface regularity of the tumor and anatomic location of tumors. As for the masses, there was a difference between groups (p=0.0077), with 0.78±1.13cm for group 1 and 1.81±2.29 cm for group 2. 40% of benign masses and 60% of malignant masses were diagnosed, according to the types of malignancies. For VEGF, the average values were 2.22±0.89 for malignant tumors and 1.66±0.91 for benign, with differences between groups (p=0.0315). The mammary tumors do not exhibit characteristics of reproductive history and clinical examination to help the differential diagnosis, and histopathology is the only method for completion of diagnosis and immunohistochemistry which can be used for injury prognosis.

Co-culture of apoptotic breast cancer cells with immature dendritic cells: a novel approach for DC-based vaccination in breast cancer

A dendritic cell (DC)-based vaccine strategy could reduce the risk of recurrence and improve the survival of breast cancer patients. However, while therapy-induced apoptosis of hepatocellular and colorectal carcinoma cells can enhance maturation and antigen presentation of DCs, whether this effect occurs in breast cancer is currently unknown. In the present study, we investigated the effect of doxorubicin (ADM)-induced apoptotic MCF-7 breast cancer cells on the activation of DCs. ADM-induced apoptotic MCF-7 cells could effectively induce immature DC (iDC) maturation. The mean fluorescence intensity (MFI) of DC maturity marker CD83 was 23.3 in the ADM-induced apoptotic MCF-7 cell group compared with 8.5 in the MCF-7 cell group. The MFI of DC co-stimulatory marker CD86 and HLA-DR were also increased after iDCs were treated with ADM-induced apoptotic MCF-7 cells. Furthermore, the proliferating autologous T-lymphocytes increased from 14.2 to 40.3% after incubated with DCs induced by apoptotic MCF-7 cells. The secretion of interferon-γ by these T-lymphocytes was also increased. In addition, cell-cell interaction between apoptotic MCF-7 cells and iDCs, but not soluble factors released by apoptotic MCF-7 cells, was crucial for the maturation of iDCs. These findings constitute a novel in vitro DC-based vaccine strategy for the treatment of breast cancer by ADM-induced apoptotic MCF-7 cells.

Analysis of CD83 antigen expression in human breast fibroadenoma and adjacent tissue / Análise da expressão do antígeno CD83 no fibroadenoma mamário humano e no tecido adjacente

CONTEXT AND OBJECTIVE: Dendritic cell maturation is considered essential for starting an immune response. The CD83 antigen is an important marker of dendritic cell maturation. The objectives here were to analyze CD83 antigen expression in human breast fibroadenoma and breast tissue adjacent to the lesion and to identify clinical factors that might influence this expression. DESIGN AND SETTING: This was a retrospective study at a public university hospital, in which 29 histopathological samples of breast fibroadenoma and adjacent breast tissue, from 28 women of reproductive age, were analyzed. METHODS: The immunohistochemistry method was used to analyze the cell expression of the antigen. The antigen expression in the cells was evaluated by means of random manual counting using an optical microscope. RESULTS: Positive expression of the CD83 antigen in the epithelial cells of the fibroadenoma (365.52; standard deviation ± 133.13) in relation to the adjacent breast tissue cells (189.59; standard deviation ± 140.75) was statistically larger (P < 0.001). Several clinical features were analyzed, but only parity was shown to influence CD83 antigen expression in the adjacent breast tissue, such that positive expression was more evident in nulliparous women (P = 0.042). CONCLUSIONS: The expression of the CD83 antigen in the fibroadenoma was positive and greater than in the adjacent breast tissue. Positive expression of the antigen in the adjacent breast tissue was influenced by parity, and was significantly more evident in nulliparous women.

CONTEXTO E OBJETIVOS: A maturação da célula dendrítica é considerada essencial para o início da resposta imune. O antígeno CD83 é um importante marcador da maturação da célula dendrítica. Os objetivos são analisar a expressão do antígeno CD83 no fibroadenoma mamário humano e no tecido mamário adjacente à lesão e identificar fatores clínicos que possam influenciar esta expressão. TIPO DE ESTUDO E LOCAL: Este é um estudo retrospectivo, realizado em um hospital público universitário, onde 29 amostras histopatológicas de fibroadenomas de mamas e de tecidos mamários adjacentes, de 28 mulheres em idade reprodutiva, foram analisados. MÉTODOS: O método de imunoistoquímica foi utilizado na análise da expressão celular do antígeno. A expressão do antígeno nas células foi avaliada por contagem aleatória e manual utilizando-se microcópio de luz. RESULTADOS: A expressão positiva do antígeno CD83 nas células epiteliais dos fibroadenomas (365,52; desvio padrão ± 133,13) em relação às células do tecido mamário adjacente (189,59; desvio padrão ± 140,75) foi estatisticamente superior (P < 0,001). Vários aspectos clínicos foram analisados, porém, a paridade se mostrou influente na expressão do antígeno CD83 no tecido mamário adjacente, onde a expressão positiva foi mais evidente nas mulheres nulíparas (P = 0,042). CONCLUSÕES: A expressão do antígeno CD83 foi positiva e mais expressiva no fibroadenoma do que no tecido mamário adjacente. A expressão positiva do antígeno no tecido mamário adjacente foi influenciada pela paridade, sendo significativamente mais evidente nas mulheres nulíparas.

Subtipos moleculares do câncer de mama / Molecular subtypes of breast cancer

O câncer de mama apresenta alta heterogeneidade clínica, morfológica e biológica, fato esse justificado pela existência de diversas formas moleculares. Diferentes perfis de expressão gênica foram caracterizados, possibilitando a identificação de subtipos moleculares distintos, com fatores prognósticos e alvos terapêuticos específicos. Esta revisão foi conduzida utilizando artigos científicos das bases de dados MEDLINE, SciELO, LILACS e PubMed e teve como objetivo discutir os subtipos moleculares do câncer de mama e suas principais características. O subtipo luminal A apresenta, com relação aos demais, o melhor prognóstico. Na sua maioria, são tumores histologicamente de baixo grau e apresentam resposta inferior à quimioterapia, enquanto, tumores luminais B apresentam maior proliferação e são, muitas vezes, de alto grau histológico. O subtipo superexpressão do receptor tipo 2 do fator de crescimento epidérmico humano (HER2), sem a terapia adjuvante sistêmica, tem menor sobrevida livre de doença e elevada taxa de recorrência, porém se beneficia de terapias alvoespecíficas. O subtipo basaloide demonstra prognóstico mais reservado, associado à menor sobrevida livre de doença e à menor sobrevida global. A anatomia patológica e o teste de imunoistoquímica, através da classificação tumoral, são de fundamental relevância na abordagem terapêutica do carcinoma mamário. Para a atual classificação molecular por imunoistoquímica, recomenda-se a adoção do painel de fatores preditivos receptor de estrogênio (RE), receptor de progesterona (RP) e HER2 para todos os casos, adicionando-se outros marcadores, como o receptor tipo 1 do fator de crescimento epidérmico (EGFR), a citoceratina 5 e o Ki-67

Breast cancer has highly heterogeneous clinical, morphological and biological features, a fact that is justified by the existence of several molecular forms. Different gene expression profiles were characterized, enabling the identification of different molecular subtypes with specific prognostic factors and therapeutic targets. This review was conducted using scientific articles of the databases MEDLINE, SciELO, LILACS and PubMed, and aimed to discuss the molecular subtypes of breast cancer and their main characteristics. The luminal A subtype, when compared with the others, features the best prognosis. Most tumors are histologically low grade and have less response to chemotherapy, while luminal B tumors have high cell proliferation rate and are most often high grade. The enriched subtype to receptor 2 human epidermal growth factor (HER2), without adjuvant systemic therapy, has a lower disease-free survival and higher recurrence rate, but it benefits from target-specific therapies. The basal-like subtype pattern shows poor prognosis associated with lower disease-free survival and shorter overall survival. The pathology and immunohistochemistry test, by tumor classification, are of fundamental importance in the therapeutic management of breast cancer. For the current molecular classification by immunohistochemistry, the adoption of the panel of the predictive factors estrogen receptors (ER), receptors progesterone (PR) and HER2 is recommended for all cases of breast cancer, adding other markers such as epidermal growth factor receptor type 1 (EGFR), the cytokeratin 5 and the Ki-67

Effects of Laughter Therapy on Depression, Quality of Life, Resilience and Immune Responses in Breast Cancer Survivors

PURPOSE: In this study, the effects of laughter therapy on levels of depression, quality of life, resilience and immune responses in breast cancer survivors were examined. METHODS: A quasi-experimental nonequivalent control group, pretest-posttest design was used. Participants (n=37) included breast cancer survivors who finished chemotheraphy and radiation therapy: 16 in the experiment group and 21 in the control group. Data were collected from August to November 2009. The experimental group participated in laughter therapy eight times, twice a week for 60 min per session. Questionnaires were used to me-asure pretest and posttest levels of depression, quality of life and resilience. A blood test was used to analyze changes in Total T cell, T helper, T suppressor, Th/Ts ratio, Total B cell, T cell/B cell ratio and NK cell for immune responses. RESULTS: The results showed that laughter therapy was effective in increasing the quality of life and resilience in breast cancer survivors. but depression and immune responses did not differ significantly between the groups. CONCLUSION: The results of the study indicate that laughter therapy may be an effective nursing intervention to improve quality of life and resilience in breast cancer survivors.

Expression of survivin and p53 proteins and their correlation with hormone receptor status in Indian breast cancer patients.

Background : In invasive ductal carcinoma (IDC), many antiapoptotic and proapoptotic genes regulate disease outcome. Hormone receptor-mediated mechanisms have also been shown to prevent apoptosis. Therefore, relations between hormone receptor status and other molecular markers need further examination. Aims : In the present study, we analyzed the expression of apoptosis-regulating genes, viz., Survivin and mutant p53, in benign breast disease (fibroadenoma) and IDC patients. Results were then correlated with hormone receptor status of the patients. Material and Methods : Paraffin-embedded tissue samples from 63 untreated female patients with IDC and 32 female patients with fibroadenoma were used. Expression of Survivin and mutant p53 was evaluated using immunohistochemical staining method. Statistical Analysis : Fisher exact test and nonparametric correlation test (Spearman rank correlation test) were performed. Results : In fibroadenoma, 53% of patients expressed Survivin and 13% of patients expressed p53 protein. Statistically significant increase in Survivin and p53 protein expression was observed in carcinoma cases. Survivin expression correlated negatively with progesterone receptor (PR) status, but its expression was independent of estrogen receptor (ER) status. p53 expression showed negative correlation with both ER and PR status. Conclusions : Increased expression of Survivin and p53 in IDC patients and correlation with hormone receptors suggest that Survivin and p53 along with hormone receptors status are likely to contribute significantly to apoptosis resistance and may serve as therapeutic target that could increase the effectiveness of conventional breast cancer therapy.

Imunofenótipo e evolução de câncer de mama: comparação entre mulheres muito jovens e mulheres na pós-menopausa / Immunophenotype and evolution of breast carcinomas: a comparison between very young and postmenopausal women

OBJETIVO: avaliar características clínicas, patológicas e moleculares de carcinomas mamários em mulheres muito jovens em comparação a tumores de mulheres na pós-menopausa. MÉTODOS: foram selecionados 106 casos de câncer de mama de mulheres jovens e 130 casos de mulheres pós-menopausa. Foram analisados dados clínicos (idade ao diagnóstico, estadiamento, ocorrência de metástases, tempo de sobrevida global e livre de doença), anátomo-patológicos (tamanho do tumor, tipo e grau histológico do tumor primário) e marcadores moleculares (receptores de estrógeno e progesterona, HER2, p53, p63, citoqueratinas 5 e 14 e EGFR) com uso da imunoistoquímica empregando microarranjo de tecido. Foi analisada a relação entre as características clínico-patológicas, imunoistoquímicas e de sobrevidas global e livre de doença. RESULTADOS: as pacientes muito jovens apresentaram maior frequência de nuliparidade (p=0,03), maior diâmetro dos tumores (p≤0,000), estadiamento clínico mais avançado (p=0,01), maior número de linfonodos positivos (p=0,001) e tumores pouco diferenciados (p=0,004). A maioria das pacientes jovens recebeu tratamento com quimioterapia (90,8%) e radioterapia (85,2%) e em menor proporção com tamoxifeno (31,5%), comparado às mulheres na pós-menopausa. Observamos baixa positividade para o receptor de estrógeno (49,1%; p=0,01) e alta positividade para a proteína HER2 (28,7%; p=0,03) nas mulheres jovens. O fenótipo triplo-negativo foi observado em 29,6% no grupo jovem e em 20% nas mulheres na pós-menopausa. Os tumores de fenótipo basal foram mais frequentes nas mulheres jovens (50%). As metástases sistêmicas ocorreram em 55,3% dos casos nas jovens e em 39,2% nas idosas. As sobrevidas global e livre de doença em cinco anos foram, respectivamente, 63 e 39% para as mulheres jovens e 75 e 67% para o grupo de mulheres na pós-menopausa...

PURPOSE: the objective of this study was to evaluate the clinical, pathological and molecular characteristics in very young women and postmenopausal women with breast cancer. METHODS: we selected 106 cases of breast cancer of very young women (≤35 years) and 130 cases of postmenopausal women. We evaluated clinical characteristics of patients (age at diagnosis, ethnic group, family history of breast cancer, staging, presence of distant metastases, overall and disease-free survival), pathological characteristics of tumors (tumor size, histological type and grade, axillary lymph nodes status) and expression of molecular markers (hormone receptors, HER2, p53, p63, cytokeratins 5 and 14, and EGFR), using immunohistochemistry and tissue microarray. RESULTS: when comparing clinicopathologic variables between the age groups, younger women demonstrated greater frequency of nulliparity (p=0.03), larger tumors (p≤0.000), higher stage disease (p=0.01), lymph node positivity (p=0.001), and higher grade tumors (p=0.004). Most of the young patients received chemotherapy (90.8%) and radiotherapy (85.2%) and less tamoxifen therapy (31.5%) comparing with postmenopausal women. Lower estrogen receptor positivity 49.1% (p=0.01) and higher HER2 overexpression 28.7% (p=0.03) were observed in young women. In 32 young patients (29.6%) and in 20% of the posmenopausal women, the breast carcinomas were of the triple-negative phenotype (p=0.034). In 16 young women (50%) and in 10 postmenopausal women (7.7%), the tumors expressed positivity for cytokeratin 5 and/or 14, basal phenotype (p=0.064). Systemic metastases were detected in 55.3% of the young women and in 39.2% of the postmenopausal women. Breast cancer overall survival and disease-free survival in five years were, respectively, 63 and 39% for young women and 75 and 67% for postmenopausal women. CONCLUSIONS: breast cancer arising in very young women showed negative clinicobiological characteristics and more aggressive tumors.