Introduction: Triple-negative breast cancer (TNBC) does not express estrogen and progesterone receptors, and does not overexpress the human epidermal growth factor 2. It represents 15%‒20% of breast cancers and have worse prognosis, with scarce available therapies and overall survival (OS) of 18 months. For these particularities, research on TNBC is important for its greater understanding. Objectives: To describe the clinical-epidemiological profile of patients with TNBC at Instituto Mário Penna (IMP). To compare findings with data from the literature. Methods: Consultation of breast immunohistochemistry (IHC) performed at IMP between July/2012 and June/2017. TNBC were selected. Data were collected from patients in electronic medical records. Maximum follow-up until December/2018. Database and statistical analysis using the SPSS program. Bibliographic review used the key phrase: “triple-negative breast cancer”. Results: 1,343 breast IHC performed at IMP in the studied period, 168 were TNBC (12.5%). Mean age of 53.4 years. Mean follow-up of 41.7 months. Neoadjuvant chemotherapy (CT) performed in 46.4%, with 12.8% of complete pathological response. Mean SG of 23.6 months, 20.2% progressed before the end of the treatment. Tumor mean size of 4.04 cm. Mortality of 22%, with 31.5% without information on death in the medical record, and about 17% on average with missing information. Table 1 shows the frequency distribution of the variables evaluated. Discussion: TNBC is a heterogeneous group of diseases, more commonly found in people aged under 40 years, of African descent, diagnosed at an advanced stage and with a high histological grade. Earlier metastasis, preferably visceral. More sensitive to CT, but with worse OS compared to other subtypes. Use of platinum, capecitabine and recent studies with immunotherapy are promising, in the search for better outcomes. Conclusion: The profile of patients with TNBC in IMP is compatible with that described in the literature. This study is a hypothesis generator and the basis for more complex research. High rates of missing information are a limiting factor.
Objective: The identification and characterization of miRNAs derived from microvesicles of breast cancer patients is the main goal of this study. Methods: A cohort of 28 patients was evaluated, 28.6% are HER2 overexpressed, 46.4% are triple-negative, and 25% are triple-positive. The study protocol was approved by the Ethics Committee of Instituto Mário Penna (CAEE 82703418.8.0000.5121). For this purpose, the peripheral blood was collected in EDTA tubes and obtained the red blood cell-free plasm. The microvesicles were purified from plasm using the microRNeasy kit (Qiagen) with posterior small RNA precipitation, according to manufacturer’s instructions. The small RNA obtained was used for high throughput sequencing using the QIAseq miRNA Library kit (Qiagen) for library construction, according to manufacturer’s instructions. The sequencing was performed by the Illumina NextSeq 550. The sequences obtained were filtered by quality, the adapters were removed, and small RNA patterns were evaluated using the Unitas (version 1.7.8). Results: The three groups of patients showed a significant abundance of miRNA profiles. The triple-negative breast cancer (TNBC) patients showed the highest relative abundance, which can be due to the more intense exocrine activity of this type of tumor. Furthermore, our results highlighted a great abundance of miR-223-3p in the TNBC patient group. Conclusion: Normally, TNBC patients have an aggressive condition of disease, and cell proliferation, migration, and invasion are common events. These characteristics can be regulated by miRNAs exported from tumor cells in microvesicles. Several miRNAs are already related to these events, and this makes them potential therapeutic or diagnostic targets for this disease. miR-223-3p was previously related to epithelial-mesenchymal transition, cell proliferation, and migration. This phenotypic effect is a strong indication that this miRNA could be used as a biomarker in TNBC management and opens great possibilities for further validation of this as a tool for liquid biopsy tests.
Objective: The objective of this study was to standardize the Ficoll gradient technique for the circulating hematopoietic stem cell (HSC) isolation for the assembly of the peripheral blood mononuclear cell (PBMC) biorepository of breast cancer (BC) patients attended in the Clinical Oncology Service of Instituto Mário Penna. Methods: The study protocol was approved by the Ethics Committee of Instituto Mário Penna (CAEE 82703418.8.0000.5121). In recommended protocols, 15 mL of blood was used. At first, we adapted this volume due to the limited amounts of samples for research available. Blood was collected in a 9-mL sodium heparin tube. The experiments were performed in 50-mL conical tubes, but with reduced blood volume, and no PBMC ring was formed. It was necessary to change to 15 mL conical tubes. Finally, the remaining red blood cells were lysed with ammonium chloride. However, with the reduced volume, this solution lysed the PBMC too. Then, we decided to remove this step from the protocol. Results: We obtained 8.06×106 cells/mm3 with 80% viability. Data were confirmed by a Neubauer camera and an automatic cell counter. The HSCs were labeled with antibodies against CD34 and CD133 by flow cytometry. Conclusion: The characterization of HSCs is important to link tumor-associated HSCs with malignant and immunosuppressive phenotypes. Studies are in progress with this standardization, and they will permit us to perform the HSC characterization of BC patients with a better knowledge of tumor microenvironment.
Background: Breast cancer is the most common cancer in women in Brazil and worldwide. There is large variation in survival among patients and molecular subtypes are important prognostic factors. However, most of the data comes from developed countries such as the United States and in Europe. Aim: Our goal was to describe breast cancer patients' demographic and pathologic characteristics, as well as their survival according to estimated molecular subtypes, assessed by common immunohistochemistry stains. Methods: AMAZONA study is a retrospective cohort conducted from June 2008 to January 2009 including women of at least 18 years old, with histologically proven breast cancer diagnosed in the period between 1 January 2001 and 31 December 2001 and between 1 January 2006 and 31 December. Estimated molecular subtypes by local immunohistochemical stains were luminal A, luminal B, HER-2 positive and triple-negative. Data were obtained from medical records and public databases. Kaplan-Meier method was used for data description and log-rank test for comparison between the subgroups. Results: 2296 patients were included in this analysis. Mean age was 54 years. Most subjects included came from hospitals located in the southeast region of the country, treated in the public health system and had stage II invasive ductal carcinoma of breast. Regarding subtype, 71.3% had hormonal receptor positive disease, 15.7% were HER-2 positive and 21.1% had triple-negative breast cancer. Overall survival (OS) was significantly different among molecular subtypes and was independent of pathologic stage for stages II and III patients. For stage III patients 5-years OS for luminal A subtype was 75.8% and for triple-negative was 56.1% ( P .0002). Conclusion: Classification of breast cancer patients in predicted molecular subtypes using immunohistochemistry is currently available in most underdeveloped countries and is a useful prognostic tool that goes beyond clinical or pathologic stage.
Objective: Due to the already observed importance of neutrophil-derived microvesicles (NMVs) in cancer development and progression, this study aims to quantify NMVs according to clinical staging and histological grade in the blood of breast cancer (BC) patients for a possible use in the liquid biopsy technique and personalized medicine, assisting in treatment decision. Methods: Peripheral blood was collected from 19 healthy women (control group) and from 51 patients with locally advanced BC (case group) in the Instituto Mário Penna, Belo Horizonte, Brazil. The study protocol was approved by the Ethics Committee of Instituto Mário Penna (CAEE 82703418.8.0000.5121). Clinical staging and histological grade data were obtained from the medical records of the study patients. The characterization of circulating NMVs was performed by immunophenotyping with specific neutrophil markers (CD66 and CD16), and quantification was performed by flow cytometry. Results: Our data showed a higher number of NMVs in BC patients, regardless of clinical staging and degree of tumor differentiation, when compared to the control group. Although no difference was observed in relation to the histopathological grade, the NMVs appear to have a potential diagnostic in BC patients. Conclusion: In a clinical scenario, they are going to use liquid biopsy as a candidate strategy to support clinical decision-making and guide therapeutic choices.
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