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Although peptide vaccines offer a novel venue for cancer immunotherapy, clinical success has been rather limited. Cell-penetrating peptides, due to their ability to translocate through the cell membrane, could be conjugated to the peptide vaccine to2 enhance therapeutic efficiency. The S4 transduction domain of the shaker-potassium channel was conjugated to mammaglobin-A (MamA) immunodominant epitope (MamA2.1) to verify its anticancer immunogenicity. S4-MamA2.1 peptide has demonstrated significantly higher epitope loading and stable membrane expression of HLA-A2 antigen-presenting molecules on T2 cell lines. Further, these S4-MamA2.1 treated T2 cells were able to activate naïve CD8+ T cells to induce MamA-specific cytotoxicity against breast cancer cells. Conjugation of the S4 domain has also demonstrated a slight increase in immunogenicity of lesser immunodominant MamA epitopes. The conjugation of the S4 domain to N-terminus of MamA2.1 demonstrated significantly higher immunogenicity over C-terminus conjugation. Taken together, the results of the present study suggest that conjugation of the S4 cell-penetrating peptide domain to MamA2.1 epitope enhances the peptide vaccine immunogenicity against MamA-expressing breast cancers. [ABSTRACT FROM AUTHOR]

Background/Aim. Human mammaglobin is considered to be one of the most significant markers of hematogenous dissemination of breast carcinoma. This paper aimed to indicate the important role of peritumoral tissue as an active participant in the tumorigenesis process and the concentration/expression of mammaglobins in the peritumoral tissue as a significant prognostic factor. Methods. This research included 64 female patients with primary breast carcinoma during the five-year follow- up period. To determine the concentration of mammaglobin A in samples of carcinoma tissue and peritumoral tissue, Enzyme-linked immunosorbent assay (ELISA) test was used, and for the determination of relative gene expression of mammaglobin A, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used. Results. The concentration of mammaglobin A was increased in both the carcinoma tissue and peritumoral tissue with an increase in tumor size, number of affected lymph nodes, number of metastases, while relative expression of mammaglobin A was statistically significantly higher in carcinoma tissue than in peritumoral tissue, regardless of the tumor size, number of affected lymph nodes, number of metastases and tumor type. The concentration of mammaglobin A was higher in peritumoral tissue than in tissue of ductal carcinoma, while in the case of lobular carcinoma the concentration of mammaglobin A was higher in carcinoma tissue than in peritumoral tissue. Conclusion. Mammaglobin A concentration in peritumoral tissue higher than 0.6704221 ng/mL, and in carcinoma tissue higher than 0.5784426 ng/mL, as well as mammaglobin A relative gene expression in carcinoma tissue higher than 1.003, were determined as cut-off values. These values may identify patients who are at higher risk of metastatic disease, which would be treated with early radical adjuvant treatment. [ABSTRACT FROM AUTHOR]

Highlights • Highly sensitive detection of metastatic breast cancer cells was demonstrated using terahertz chemical microscopy with aptamers as the probes. • Metastatic breast cancer cells MCF7 and MDA-MB-415 cells were detected by scanning sensing plates immobilized with MAMB1 and MAMA2 aptamers using THz radiation. Abstract We demonstrate high-sensitivity detection of metastatic breast cancer cells using terahertz (THz) chemical microscopy (TCM) with aptamers as ligands. For the aptamers, we use previously developed synthetic single stranded (ss) DNA aptamers mammaglobin B1 (MAMB1) and mammaglobin A2 (MAMA2) that bind to mammaglobin B and mammaglobin A proteins, respectively, which are overexpressed on the surface of MCF7 and MDA-MB-415 breast cancer cells. Each aptamer was immobilized on the surface of a sensing plate, and the amplitude of the THz signal was measured upon the binding of each aptamer to different number (10–106) of its target breast cancer cells. A change in the THz amplitude was observed when MAMB1 and MAMA2 bind to MCF7 and MDA-MB-415, respectively. We find that this change was linear as a function of the log number of breast cancer cells used. No change in the THz amplitude was observed when the same number of normal breast cells (MCF 10A) were used. Moreover, MAMB1 and MAMA2 did not show binding to the counter breast cancer cells, indicating high selectivity. We have demonstrated that the TCM using aptamers as ligands has a limit of detection as small as 1 breast cancer cell in 100 μL of sample. Results described here indicate that the TCM could be a powerful tool to detect metastatic breast cancer cells. [ABSTRACT FROM AUTHOR]

Aims: In addition to oestrogen and progesterone receptors, gross cystic disease fluid protein-15 (GCDFP-15) and mammaglobin A (MAM) are the most common markers used to identify breast origin by immunohistochemistry. GCDFP-15 expression has been reported in approximately 60% of breast carcinomas and MAM expression in approximately 80%. Data on their expression in triple-negative breast cancer (TNBC) are very limited. The aim of this study was to examine the expression of these markers in TNBC to determine their utility in pathological diagnosis. Methods and results: We studied the immunohistochemical (IHC) expression of GCDFP-15 and MAM in 63 primary and 118 metastatic TNBCs. GCDFP-15 staining was present in 14% of primary and 21% of metastatic TNBCs. MAM staining was present in 25% of primary and 41% of metastatic TNBCs. The frequency of expression of GCDFP-15 and/or MAM was 30% in primary and 43% in metastatic TNBCs, and many positive tumours had only focal staining. Conclusions: Staining for GCDFP-15 and/or MAM in triple-negative carcinomas helps to confirm breast origin, but most tumours in this subgroup of breast carcinomas lack expression of either marker. [ABSTRACT FROM AUTHOR]

Abstract: Objective: A one-tube nested RT-PCR protocol was set up and used to detect mammaglobin A (MGA) expression in blood samples from breast cancer patients. The correlation of MGA detection with prognostic factors was analyzed. Design and methods: Total RNA from nucleated blood cells was extracted from 65 breast cancer patients (before surgery and after the treatments) and 18 healthy subjects and used to detect MGA expression by a modified nested RT-PCR. Results: MGA expression was detected in 38.4% of patients before surgery, and in 50% and 36.8% of post-treatment samples from patients that expressed MGA or were MGA negative before surgery, respectively. MGA detection was associated with the absence of tumor estrogen receptors (p =0.004). Conclusions: MGA detection by the modified nested RT-PCR is a specific marker for circulating tumor cells in patients with breast carcinoma and a negative prognostic factor for the disease. [Copyright &y& Elsevier]

Abstract: Objectives: : The development and validation of a nested RT-PCR methodology for the detection of Mammaglobin A-mRNA-positive circulating tumor cells in peripheral blood of patients with operable breast cancer and evaluation of its prognostic significance. Design and methods: : Different combinations of specific primers were in silico designed and selected, so that false positive results due to genomic DNA contamination were avoided. The specificity of the primers used was evaluated in 30 healthy individuals, 20 patients with colorectal cancer and 20 patients with non-small cell lung cancer. The method was applied in 101 patients with operable breast cancer before the administration of adjuvant chemotherapy and 39 patients with metastatic breast cancer. Results: : Mammaglobin A-mRNA-positive cells were detected in 14/101 (13.9%) of early breast cancer patients but not in the control population studied (0%); 9 of them (64.3%) relapsed during the follow-up period. Mammaglobin A was detected in 7/39 (17.9%) of patients with verified metastasis. Multivariate analysis revealed the detection of Mammaglobin A-mRNA-positive cells, as an independent risk factor for reduced DFI. Conclusions: : Mammaglobin A is a highly specific molecular marker for the detection of circulating tumor cells in operable breast cancer, with important prognostic applications. [Copyright &y& Elsevier]