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PurposeTriple-negative breast cancer (TNBC), an aggressive breast cancer subtype, is genetically heterogeneous which challenges the identification of clinically effective molecular makers. Extracellular vesicles (EVs) are key players in the intercellular signaling communication and have been shown to be involved in tumorigenesis. The main goal of this study was to evaluate the role and mechanisms of EVs derived from TNBC cells in modulating proliferation and cytotoxicity to chemotherapeutic agents in non-tumorigenic breast cells (MCF10A).MethodsEVs were isolated from TNBC cell lines and characterized by nanoparticle tracking analysis, Western blot, and transmission electron microscopy. MCF10A cells were treated with the isolated EVs and evaluated for cell proliferation and cytotoxicity to Docetaxel and Doxorubicin by the MTT and MTS assays, respectively. Gene and miRNA expression profiling was performed in the treated cells to determine expression changes that may be caused by EVs treatment.ResultsMCF10A cells treated with HCC1806-EVs (MCF10A/HCC1806-EVs) showed a significant increase in cell proliferation and resistance to the therapeutic agents tested. No significant effects were observed in the MCF10A cells treated with EVs derived from MDA-MB-231 cells. Gene and miRNA expression profiling revealed 138 genes and 70 miRNAs significantly differentially expressed among the MCF10A/HCC1806-EVs and the untreated MCF10A cells, affecting mostly the PI3K/AKT, MAPK, and HIF1A pathways.ConclusionEVs isolated from the HCC1806 TNBC cells are capable of inducing proliferation and drug resistance on the non-tumorigenic MCF10A breast cells, potentially mediated by changes in genes and miRNAs expression associated with cell proliferation, apoptosis, invasion, and migration.Electronic supplementary materialThe online version of this article (10.1007/s10549-018-4925-5) contains supplementary material, which is available to authorized users.

MicroRNAs derived from extracellular vesicles (EV-miRNAs) are circulating miRNAs considered as potential new diagnostic markers for cancer that can be easily detected in liquid biopsies. In this study, we performed RNA sequencing analysis as a screening strategy to identify EV-miRNAs derived from serum of clinically well-annotated breast cancer (BC) patients from the south of Brazil. EVs from three groups of samples (healthy controls (CT), luminal A (LA), and triple-negative (TNBC)) were isolated from serum using a precipitation method and analyzed by RNA-seq (screening phase). Subsequently, four EV-miRNAs (miR-142-5p, miR-150-5p, miR-320a, and miR-4433b-5p) were selected to be quantified by quantitative real-time PCR (RT-qPCR) in individual samples (test phase). A panel composed of miR-142-5p, miR-320a, and miR-4433b-5p distinguished BC patients from CT with an area under the curve (AUC) of 0.8387 (93.33% sensitivity, 68.75% specificity). The combination of miR-142-5p and miR-320a distinguished LA patients from CT with an AUC of 0.9410 (100% sensitivity, 93.80% specificity). Interestingly, decreased expression of miR-142-5p and miR-150-5p were significantly associated with more advanced tumor grades (grade III), while the decreased expression of miR-142-5p and miR-320a was associated with a larger tumor size. These results provide insights into the potential application of EVs-miRNAs from serum as novel specific markers for early diagnosis of BC.

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Improvement of breast cancer (BC) patient's outcome is directly related to early detection. However, there is still a lack of reliable biomarkers for diagnosis, prognosis and, treatment follow up in BC, leading researchers to study the potential of liquid biopsy based on circulating microRNAs (c-miRNAs). These c-miRNAs can be cellfree or associated with extracellular vesicles (EVs), and have great advantages such as stability in biofluids, noninvasive accessibility compared to current techniques (core-biopsy and surgery), and expression associated with pathogenic conditions. Recently, a new promising field of EV-derived miRNAs (EV-miRNAs) as cancer biomarkers has emerged, receiving special attention due to their selective vesicle sorting which makes them accurate for disease detection. In this review, we discuss new findings about c-miRNA and their potential as biomarkers for BC diagnosis, prognosis, and therapy. Additionally, we address the impact of limitations associated with the standardization of analysis techniques and methods on the implementation of these biomarkers in the clinical setting. (Translational Research 2020; 223:40À60) Abbreviations: BC = breast cancer; c-miRNAs = circulating microRNAs; cf-miRNAs = cell-free microRNAs; EVs = extracellular vesicles; EV-miRNAs = EV-derived miRNAs; ER = estrogen receptor; PR = progesterone receptor; ERBB2 = human epidermal growth factor receptor 2; TNBC = triple negative breast cancer; CTCs = circulating tumor cells; CA15-3 = cancer antigen 15-3; CEA = carcinoembryonic antigen; RANTES/CCL5 = chemokine (CÀC motif) ligand 5; OPN = osteopontin; PAI-1 = plasminogen activator inhibitor-1; AUC = area under the curve; RT-qPCR = reverse transcription quantitative polymerase chain reaction; CT = controls; OS = overall survival; dPCR = digital PCR; NGS = next generation sequencing BACKGROUND Breast cancer (BC) is the most frequently diagnosed cancer in females and the leading cause of death by cancer worldwide, with the mortality rate predicted to reach around 626,679 deaths worldwide in 2018. 1 This cancer can be classified in several ways, one of the methods of clinical evaluation being the immunohistochemistry classification. 2 According to this classification, BC is stratified based on the expression of 4 main protein markers À estrogen receptor (ER), progesterone, receptor (PR), the proliferation marker and

Chemokines and its receptors have significant impact on physiological and pathological processes and studies concerning their association with tumor biology are subject of great interest in scientific community. CXCL12/CXCR4 axis has been widely studied due to its significant role in tumor microenvironment, but it is also important to development and maintenance of tissues and organs, for example, in the brain and cerebellum. Studies have demonstrated that CXCL12 and CXCR4 are required for normal cerebellar development and that dysfunction in this pathway may be involved with medulloblastoma pathogenesis. In this context, a new molecular subgroup has been suggested based on the importance of the association between CXCR4 overexpression and sonic hedgehog subgroup. Treatment using CXCR4 antagonists showed significant results, evidencing the important role and possible therapeutic capacity of CXCR4 in MB. This review summarizes studies on MB cell biology, focusing on a chemokine-receptor axis, CXCL12/CXCR4, that may have implications for treatment strategies once it can improve life expectancy and reduce neurocognitive sequelae of patients with this neoplasia.Tumors originate from cells with dysregulation on normal growth mechanism control, caused by genetic mutations. 1 Studies have demonstrated that the tumor microenvironment is constituted not only by tumor cells, but also of extracellular matrix, fibroblasts, endothelial cells and immune cells that could influence tumor progression. 2,3 One chemokine that has been acquiring relevance in cancer is CXCL12, whose receptor CXCR4 is overexpressed in at least 20 different human cancers, including breast cancer, ovarian cancer, melanoma and prostate cancer. 4 Besides their critical role in tumor cell growth, 5 survival and angiogenesis in multiple cancers, 6 this chemokine receptor has been described as an important homing and metastatic mediator of secondary growth in organs that produce CXCL12, such as liver, 7 lung and bone marrow. 8 However, the contribution of CXCR4/CXCL12 axis in organ-specific dissemination and tumor growth has been strongly debated. [9][10][11] Studies have shown that CXCR4, a molecule strongly expressed in proliferating granule neuron precursors (GNP) that are cell types associated with medulloblastoma (MB), 12 is also involved with sonic hedgehog (SHH) pathway, 13 as well as its ligand, that significantly enhances SHH-induced cell proliferation. 14,15 CXCR4 is predominantly expressed in tumor areas, while CXCL12 is expressed mainly in the endothelium of tumor associated blood vessels. 16,17 It has also been described another receptor for the chemokine CXCL12, the CXCR7, however, it seems to play no role in this tumor. 15 CXCL12 and CXCR4 are expressed in several brain tumors, including MB and the expression level of this receptor appears to have prognostic significance. 16,18,19 Positive CXCR4 expression was identified in nine of ten samples of MB tumors, in contrast to little or no staining in normal cerebellum counterparts. 16 Studies als...

BackgroundWilms’ tumor is an embryonal neoplasm of the kidney that accounts for approximately 6 % of all childhood tumors. The chemokine CXCL12 (C-X-C chemokine ligand 12) and its ligand CXCR4 (C-X-C chemokine receptor type 4) are involved in the development of several organs, including the kidney, and are also associated with tumor growth and metastasis. FOXP3 (forkhead transcription factor 3) was initially described as a marker for regulatory T cells; however, its expression in several types of tumor cells has already been described and may have prognostic significance. The aim of the present study was to analyze rs3761548 and rs2232365 FOXP3 polymorphisms, as well as evaluate rs1801157 CXCL12 polymorphism in Wilms’ tumor samples.MethodsPolymorphisms were evaluated in 32 patients and 78 neoplasia-free controls. Genotypes of rs1801157 were determined using PCR-restriction fragment length polymorphism (PCR-RFLP) method, and genotypes of rs2232365 and rs3761548 were determined using allele-specific PCR (AS-PCR).ResultsThe case-control study indicated a significant association for allele A carriers of rs1801157 polymorphism in relation to Wilms’ tumor susceptibility (OR = 5.261; 95 % CI 2.156 to 12.84; p = 0.0002). The opposite was observed in male carriers of G allele for rs2232365 polymorphism (OR 0.1164; 95 % CI 0.0227 to 0.5954; p = 0.0091) or when male and female subjects were analyzed (OR = 0.1304; 95 % CI 0.05013 to 0.3394; p < 0.0001).ConclusionsAll in all, these markers may contribute to this neoplasia susceptibility and progression; however, further studies are needed to real clarify their role in Wilms’ tumor pathogenesis.

Breast cancer represents a complex and heterogeneous disease that comprises distinct disease conditions, histological features, and clinical outcome. Since many years, it has been demonstrated as an association between HER2 amplification and poor prognosis, because its overexpression is associated with an aggressive phenotype of breast tumor cells. A significant proportion of cases have developed resistance to the current therapies available. Consequently, new prognostic markers are urgently needed to identify patients who are at the highest risk for developing metastases. During the past decade, new insights provided valuable knowledge regarding mechanisms underlying the dynamic interplayed between immune cells and tumor progression. It has been shown that the presence of a lymphocytic infiltrate, particularly of regulatory T cells, in cancer tissue, is associated with clinical outcome promoting rather than inhibiting cancer development and progression. It has been also verified that the clinical value of lymphocytic infiltration in breast cancers could be subtype-dependent, including the HER2-enriched subtype. In this context, this work summarizes proposed to discuss the prognostic value of regulatory T cell infiltration in microenvironment of HER2-enriched breast cancer.