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HER2 is a well-studied tyrosine kinase (TK) membrane receptor which functions as a therapeutic target in invasive ductal breast carcinomas (IDC). The standard of care for the treatment of HER2-positive breast is the antibody trastuzumab. Despite specific treatment unfortunately, 20% of primary and 70% of metastatic HER2 tumors develop resistance. HER2 belongs to a gene family, with four members (HER1-4) and these members could be involved in resistance to anti-HER2 therapies. In this study we designed a probemix to detect the amplification of the four HER oncogenes in a single reaction. In addition, we developed a protocol based on the combination of MLPA with ddPCR to detect the tumor proportion of co-amplified HERs. On 111 IDC, the HER2 MLPA results were validated by FISH (Adjusted r 2 = 0,91, p < 0,0001), CISH (Adjusted r 2 = 0,938, p < 0,0001) and IHC (Adjusted r 2 = 0,31, p < 0,0001). HER1-4 MLPA results were validated by RT-qPCR assays (Spearman Rank test p < 0,05). Of the 111 samples, 26% presented at least one HER amplified, of which 23% showed co-amplifications with other HERs. The percentage of cells with HER2 co-amplified varied among the tumors (from 2-72,6%). Independent in-silico findings show that the outcome of HER2+ patients is conditioned by the status of HER3 and HER4. Our results encourage further studies to investigate the relationship with patient's response to single or combined treatment. The approach could serve as proof of principle for other tumors in which the HER oncogenes are involved.
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Introducción El Carcinoma Lobulillar Invasor (cli) es el tipo histológico especial más común del cáncer de mama. Presenta características histopatológicas asociadas a buen pronóstico, pero algunos estudios sugieren que los resultados a largo plazo pueden ser peores que los del Carcinoma Ductal Invasor (cdi). Objetivo Los objetivos principales del estudio fueron evaluar las características clínico-patológicas del cli y establecer el valor pronóstico. Material y método Se seleccionaron 244 pacientes con cli y se utilizó como grupo control a 524 pacientes con cdi, comparándolas con relación 2 a 1. Resultados No se observaron diferencias en edad, estado menopáusico, motivo de consulta e invasión linfovascular. Fueron más frecuentemente multifocales, multicéntricos, de mayor tamaño, bajo grado histológico y her2 negativo. La cirugía conservadora se realizó con menos frecuencia. No hubo diferencias significativas en recaída a distancia, cáncer de mama contralateral, sobrevida libre de enfermedad y global. Conclusiones Las pacientes con cli no tuvieron mejores resultados a pesar de un fenotipo biológico más favorable. La histología ductal o lobulillar no debería ser un factor en el manejo de la patología, y no debería considerarse un factor pronóstico o predictivo determinante al momento del diagnóstico
Introduction Invasive Lobular Carcinoma (ilc) is the second most common histologic type of breast cancer. Typically, displays features associated with a good prognosis, but some studies suggest that outcomes of ilc may be worse than for Invasive Ductal Carcinoma (idc). Objective The main purpose of this study was to evaluate the clinical-pathological characteristics of Lobular Breast Carcinoma and establish his prognostic value. Materials and method We selected a group of 244 patients with ilc and compared with 524 patients whit idc in relation 2:1. Results There were no differences in age, menopausal status, symptoms at time of diagnosis, and lymph vascular invasion. ilc were larger, low histological grade and her2 negative, more often mulfifocal and multicentric. Breast-preservation therapy was less frequent for Invasive Lobular Carcinoma. Distant relapse, contralateral cancer, overall survival, disease-free survival, did not differ between idc and ilc. Conclusions Women with ilc do not have better clinical outcomes than patients with idc, despite the fact that the biologic phenotype of ilc is quite favorable. The ductal or lobular histology should not be a factor in the therapeutic decision-making process, and should not be considered an important prognostic or predictive factor at diagnosis
Assuntos
Neoplasias da Mama , Carcinoma Lobular , Carcinoma DuctalRESUMO
AIM: Accumulated evidence suggests that aberrant methylation of the TP73 gene and increased levels of ΔNp73 in primary tumours correlate with poor prognosis. However, little is known regarding the transcriptional and functional regulation of the TP73 gene in breast cancer. The aim of the present study was to determine the expression of the ΔNp73 isoform, its relationship with DNA methylation of TP73 and their clinical prognostic significance in breast cancer patients. METHODS: TP73 gene methylation was studied in TCGA datasets and in 70 invasive ductal breast carcinomas (IDCs). The expression of p73 isoforms was evaluated by immunohistochemistry (IHC) and Western blot and correlated with clinicopathological variables and clinical outcome. RESULTS: We observed that the methylation of diverse CpG islands of TP73 differed significantly between molecular subtypes. An inverse correlation was found between p73 protein expression and the methylation status of the TP73 gene. The expression of exon 3' of p73 (only expressed in ΔNp73) was significantly higher in patients with wild-type p53. Immunohistochemical analysis revealed that all p73 isoforms were localised in both the nuclear and cytoplasmic compartments. We confirmed a positive association between the expression of ∆Np73 and high histological grade. CONCLUSIONS: Our findings suggest that high expression of ΔNp73 could be used to determine the aggressiveness of IDCs and could be incorporated in the pathologist's report.
Assuntos
Neoplasias da Mama/diagnóstico , Carcinoma Ductal de Mama/diagnóstico , Ilhas de CpG/genética , Proteína Tumoral p73/genética , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/metabolismo , Carcinoma Ductal de Mama/patologia , Metilação de DNA , Intervalo Livre de Doença , Feminino , Humanos , Imuno-Histoquímica , Prognóstico , Isoformas de Proteínas , Proteína Tumoral p73/metabolismoRESUMO
During the last decades it has been established that breast cancer arises through the accumulation of genetic and epigenetic alterations in different cancer related genes. These alterations confer the tumor oncogenic abilities, which can be resumed as cancer hallmarks (CH). The purpose of this study was to establish the methylation profile of CpG sites located in cancer genes in breast tumors so as to infer their potential impact on 6 CH: i.e. sustained proliferative signaling, evasion of growth suppressors, resistance to cell death, induction of angiogenesis, genome instability and invasion and metastasis. For 51 breast carcinomas, MS-MLPA derived-methylation profiles of 81 CpG sites were converted into 6 CH profiles. CH profiles distribution was tested by different statistical methods and correlated with clinical-pathological data. Unsupervised Hierarchical Cluster Analysis revealed that CH profiles segregate in two main groups (bootstrapping 90-100%), which correlate with breast laterality (p = 0.05). For validating these observations, gene expression data was obtained by RealTime-PCR in a different cohort of 25 tumors and converted into CH profiles. This analyses confirmed the same clustering and a tendency of association with breast laterality (p = 0.15). In silico analyses on gene expression data from TCGA Breast dataset from left and right breast tumors showed that they differed significantly when data was previously converted into CH profiles (p = 0.033). We show here for the first time, that breast carcinomas arising on different sides of the body present differential cancer traits inferred from methylation and expression profiles. Our results indicate that by converting methylation or expression profiles in terms of Cancer Hallmarks, it would allow to uncover veiled associations with clinical features. These results contribute with a new finding to the better understanding of breast tumor behavior, and can moreover serve as proof of principle for other bilateral cancers like lung, testes or kidney.
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Neoplasias da Mama/genética , Neoplasias da Mama/fisiopatologia , Carcinoma/genética , Carcinoma/fisiopatologia , Ilhas de CpG , Adulto , Idoso , Estudos de Coortes , Metilação de DNA , Epigênese Genética , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Pessoa de Meia-IdadeRESUMO
Breast cancer is a heterogeneous disease characterized by the accumulation of genetic and epigenetic alterations that contribute to the development of regional and distant metastases. Lymph node metastasis (LNM) status is the single most important prognostic factor. Metastatic cancer cells share common molecular alterations with those of the primary tumor, but in addition, they develop distinct changes that allow the cancer to progress. There is an urgent need for molecular studies which focus on identifying genomic and epigenomic markers that can predict the progression to metastasis. The objective of this study was to identify epigenetic similarities and differences between paired primary breast tumor (PBT) and LNM. We employed Methylation-Specific-MLPA (Multiplex ligation-dependent probe amplification) to assess the methylation status of 33 cancer-related genes in a cohort of 50 paired PBT and LNM specimens. We found that the methylation index, which represents the degree of aberrantly methylated genes in a specimen, was maintained during the progression to LNM. However, some genes presented differential methylation profiles. Interestingly, PAX6 presented a significant negative correlation between paired PBT and LNM (p = 0.03), which indicated a switch from methylated to unmethylated status in the progression from PBT to LNM. We further identified that the methylation status of PAX6 on the identified CpG site functionally affected the expression of PAX6 at the mRNA level. Our study unraveled significant epigenetic changes during the progression from PBT to LNM, which may contribute to improved prognosis, prediction and therapeutic management of metastatic breast cancer patients.
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Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Epigênese Genética , Regulação Neoplásica da Expressão Gênica , Metástase Linfática , Biomarcadores Tumorais , Mama/patologia , Estudos de Coortes , Ilhas de CpG , Metilação de DNA , Progressão da Doença , Proteínas do Olho/metabolismo , Feminino , Proteínas de Homeodomínio/metabolismo , Humanos , Linfonodos/patologia , Fator de Transcrição PAX6 , Fatores de Transcrição Box Pareados/metabolismo , Prognóstico , Proteínas Repressoras/metabolismoRESUMO
In a recent study, we have shown that in mammary tumors from mice lacking the Cav-1 gene, there are alterations in specific heat shock proteins as well as in tumor development. With this in mind, we have now investigated other proteins in the same mammary mouse tumor model (Her-2/neu expressing mammary tumors from Cav-1 wild type and Cav-1 null mice), to further comprehend the complex tumor-stroma mechanisms involved in regulating stress responses during tumor development. In this tumor model the cancer cells always lacked of Cav-1, so the KO influenced the Cav-1 in the stroma. By immunohistochemistry, we have found a striking co-expression of ß-catenin and Her-2/neu in the tumor cells. The absence of Cav-1 in the tumor stroma had no effect on expression or localization of ß-catenin and Her-2/neu. Both proteins appeared co-localized at the cell surface during tumor development and progression. Since Her-2/neu activation induces MTA1, we next evaluated MTA1 in the mouse tumors. Although this protein was found in numerous nuclei, the absence of Cav-1 did not alter its expression level. In contrast, significantly more PTEN protein was noted in the tumors lacking Cav-1 in the stroma, with the protein localized mainly in the nuclei. P-Akt levels were relatively low in tumors from both Cav-1 WT and Cav-1 KO mice. There was also an increase in nuclear NHERF1 expression levels in the tumors arising from Cav-1 KO mice. The data obtained in the MMTV-neu model are consistent with a role for Cav-1 in adjacent breast cancer stromal cells in modulating the expression and localization of important proteins implicated in tumor cell behavior.
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Caveolina 1/metabolismo , Neoplasias Mamárias Animais/metabolismo , Vírus do Tumor Mamário do Camundongo/genética , PTEN Fosfo-Hidrolase/metabolismo , Fosfoproteínas/metabolismo , Receptor ErbB-2/metabolismo , Trocadores de Sódio-Hidrogênio/metabolismo , beta Catenina/metabolismo , Animais , Caveolina 1/genética , Feminino , Humanos , Imuno-Histoquímica , Células MCF-7 , Neoplasias Mamárias Animais/patologia , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor ErbB-2/genética , beta Catenina/genéticaRESUMO
Breast carcinogenesis is a multistep process that involves both genetic and epigenetic alterations. Identification of aberrantly methylated genes in breast tumors and their relation to clinical parameters can contribute to improved diagnostic, prognostic, and therapeutic decision making. Our objective in the present study was to identify the methylation status of 34 cancer-involved genes in invasive ductal carcinomas (IDC). Each of the 70 IDC cases analyzed had a unique methylation profile. The highest methylation frequency was detected in the WT1 (95.7%) and RASSF1 (71.4%) genes. Hierarchical cluster analysis revealed three clusters with different distribution of the prognostic factors tumor grade, lymph node metastasis, and proliferation rate. Methylation of TP73 was associated with high histological grade and high proliferation rate; methylation of RARB was associated with lymph node metastasis. Concurrent methylation of TP73 and RARB was associated with high histological grade, high proliferation rate, increased tumor size, and lymph node metastasis. Patients with more than six methylated genes had higher rates of relapse events and cancer deaths. In multivariate analysis, TP73 methylation and the methylation index were associated with disease outcome. Our results indicate that methylation index and methylation of TP73 and/or RARB are related to unfavorable prognostic factors in patients with IDC. These epigenetic markers should be validated in further studies to improve breast cancer management.
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Neoplasias da Mama/genética , Carcinoma Ductal de Mama/genética , Ilhas de CpG , Metilação de DNA , Regulação Neoplásica da Expressão Gênica , Adulto , Mama/metabolismo , Mama/patologia , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Carcinoma Ductal de Mama/diagnóstico , Carcinoma Ductal de Mama/patologia , Análise por Conglomerados , Proteínas de Ligação a DNA/genética , Feminino , Humanos , Pessoa de Meia-Idade , Proteínas Nucleares/genética , Prognóstico , Receptores do Ácido Retinoico/genética , Proteína Tumoral p73 , Proteínas Supressoras de Tumor/genética , Proteínas WT1/genéticaRESUMO
En la Argentina el cáncer de mama con un 25 por ciento del total es la neoplasia maligna más común en las mujeres. El objetivo del presente trabajo es describir el perfil epidemiológico, clínico y patológico de mujeres con cáncer de mama concurrentes a centros asistenciales, públicos y privados del país, adheridos voluntariamente al Grupo Colaborativo durante el período 2012-2013. De los 607 casos reportados corresponden un 27 por ciento al sector público y un 73 por ciento al privado. Según subsectores: de obra social, 63.7 por ciento; sin cobertura, 19.7 por ciento; sistema prepago, 14.2 por ciento y mutal, 2.2 por ciento. Los casos por jurisdicciones fueron: Mendoza, 20.5 por ciento; CABA, 19.9 por ciento; Buenos Aires, 13.6 por ciento; Tucumán, 10.8 por ciento; Córdoba, 6.9 por ciento ; Río Negro, 4.9 por ciento y el resto del país, 23 por ciento. Edad de presentación promedio fue de 57.5 años, Con un rango de 63.4 (90.6-27.9). La mayor frecuencia se dio entre los 50-59 años con un 27.2 por ciento. Estadios: El pTNM de presentación en 506 casos fue: estadio 0: 5.9 por ciento; I: 37.7 por ciento; II: 35.7 por ciento; III: 18.8 por ciento y IV: 1.5 por ciento. Un estudio epidemiológico colaborativo sobre el cáncer de mama brinda información sobre las características biológicas y epidemiológicas en un breve lapso, considerando diferencias geográficas, socio-demográficas, biológicas e institucionales subyacentes en la prevención y el diagnóstico de esta patología.
In Argentina breast cancer with 25 percent of the total is the most common malignancy in women. The aim of this paper is to describe the epidemiological, clinical and pathologic women attending breast cancer care centers, public an private, voluntarily adhered to a Collaborative Group for the period 2012-2013. Of the 607 cases reported 27 percent are from the public and 73 percent from private sector. According to the subsectors they were from : social work, 63.7 percent; uninsured, 19.7 percent; prepaid system, 14.2 percent and mutual, 2.2 percent. Cases by jurisdictions were: Mendoza, 20.5 percent; CABA, 19.9 percent; Buenos Aires, 13.6 percent; Tucumán, 10.8 percent; Córdoba, 6.9 percent ; Río Negro, 4.9 percent and rest of the country, 23 percent. Average age at presentation was 57.5 years, with a range 63.4 years (90.6-27.9). The highest frequency was between 50-59 years with 27.2 percent. The pTNM of 506 cases was: stage 0: 5.9 percent; I: 37.7 percent; II: 35.7 percent; III: 18.8 percent and IV: 1.5 percent. A colaborative epidemiology study on breast cancer provides information on the biological and epidemiology in a short time, considering geographical, socio-demographic, biological and institutional differences underlying the prevention and diagnosis of this pathology.
Assuntos
Humanos , Feminino , Adulto , Pessoa de Meia-Idade , Argentina/epidemiologia , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/epidemiologia , Neoplasias da Mama/mortalidade , Autoexame de Mama , Fatores Etários , Fatores de Risco , Hospitais Privados , Hospitais Públicos , Incidência , Mamografia , Sensibilidade e EspecificidadeRESUMO
En la Argentina el cáncer de mama con un 25 por ciento del total es la neoplasia maligna más común en las mujeres. El objetivo del presente trabajo es describir el perfil epidemiológico, clínico y patológico de mujeres con cáncer de mama concurrentes a centros asistenciales, públicos y privados del país, adheridos voluntariamente al Grupo Colaborativo durante el período 2012-2013. De los 607 casos reportados corresponden un 27 por ciento al sector público y un 73 por ciento al privado. Según subsectores: de obra social, 63.7 por ciento; sin cobertura, 19.7 por ciento; sistema prepago, 14.2 por ciento y mutal, 2.2 por ciento. Los casos por jurisdicciones fueron: Mendoza, 20.5 por ciento; CABA, 19.9 por ciento; Buenos Aires, 13.6 por ciento; Tucumán, 10.8 por ciento; Córdoba, 6.9 por ciento ; Río Negro, 4.9 por ciento y el resto del país, 23 por ciento. Edad de presentación promedio fue de 57.5 años, Con un rango de 63.4 (90.6-27.9). La mayor frecuencia se dio entre los 50-59 años con un 27.2 por ciento. Estadios: El pTNM de presentación en 506 casos fue: estadio 0: 5.9 por ciento; I: 37.7 por ciento; II: 35.7 por ciento; III: 18.8 por ciento y IV: 1.5 por ciento. Un estudio epidemiológico colaborativo sobre el cáncer de mama brinda información sobre las características biológicas y epidemiológicas en un breve lapso, considerando diferencias geográficas, socio-demográficas, biológicas e institucionales subyacentes en la prevención y el diagnóstico de esta patología. (AU)
In Argentina breast cancer with 25 percent of the total is the most common malignancy in women. The aim of this paper is to describe the epidemiological, clinical and pathologic women attending breast cancer care centers, public an private, voluntarily adhered to a Collaborative Group for the period 2012-2013. Of the 607 cases reported 27 percent are from the public and 73 percent from private sector. According to the subsectors they were from : social work, 63.7 percent; uninsured, 19.7 percent; prepaid system, 14.2 percent and mutual, 2.2 percent. Cases by jurisdictions were: Mendoza, 20.5 percent; CABA, 19.9 percent; Buenos Aires, 13.6 percent; Tucumán, 10.8 percent; Córdoba, 6.9 percent ; Río Negro, 4.9 percent and rest of the country, 23 percent. Average age at presentation was 57.5 years, with a range 63.4 years (90.6-27.9). The highest frequency was between 50-59 years with 27.2 percent. The pTNM of 506 cases was: stage 0: 5.9 percent; I: 37.7 percent; II: 35.7 percent; III: 18.8 percent and IV: 1.5 percent. A colaborative epidemiology study on breast cancer provides information on the biological and epidemiology in a short time, considering geographical, socio-demographic, biological and institutional differences underlying the prevention and diagnosis of this pathology. (AU)
Assuntos
Humanos , Adulto , Feminino , Pessoa de Meia-Idade , Idoso , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/epidemiologia , Neoplasias da Mama/mortalidade , Argentina/epidemiologia , Autoexame de Mama , Mamografia , Sensibilidade e Especificidade , Incidência , Fatores de Risco , Fatores Etários , Hospitais Privados , Hospitais PúblicosRESUMO
INTRODUCTION: Giant fibroadenoma is an uncommon variant of benign breast lesions. Aberrant methylation of CpG islands in promoter regions is known to be involved in the silencing of genes (for example, tumor-suppressor genes) and appears to be an early event in the etiology of breast carcinogenesis. Only hypermethylation of p16INK4a has been reported in non-giant breast fibroadenoma. In this particular case, there are no previously published data on epigenetic alterations in giant fibroadenomas. Our previous results, based on the analysis of 49 cancer-related CpG islands have confirmed that the aberrant methylation is specific to malignant breast tumors and that it is completely absent in normal breast tissue and breast fibroadenomas. CASE PRESENTATION: A 13-year-old Hispanic girl was referred after she had noted a progressive development of a mass in her left breast. On physical examination, a 10 × 10 cm lump was detected and axillary lymph nodes were not enlarged. After surgical removal the lump was diagnosed as a giant fibroadenoma. Because of the high growth rate of this benign tumor, we decided to analyze the methylation status of 49 CpG islands related to cell growth control. We have identified the methylation of five cancer-related CpG islands in the giant fibroadenoma tissue: ESR1, MGMT, WT-1, BRCA2 and CD44. CONCLUSION: In this case report we show for the first time the methylation analysis of a giant fibroadenoma. The detection of methylation of these five cancer-related regions indicates substantial epigenomic differences with non-giant fibroadenomas. Epigenetic alterations could explain the higher growth rate of this tumor. Our data contribute to the growing knowledge of aberrant methylation in breast diseases. In this particular case, there exist no previous data regarding the role of methylation in giant fibroadenomas, considered by definition as a benign breast lesion.