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American Journal of Clinical Pathology Apr 2011We investigated the expression of gross cystic disease fluid protein 15 (GCDFP) and mammaglobin (MGB) by immunohistochemical analysis in 71 invasive breast carcinomas...
We investigated the expression of gross cystic disease fluid protein 15 (GCDFP) and mammaglobin (MGB) by immunohistochemical analysis in 71 invasive breast carcinomas (IBCs) subtyped into luminal (A and B), HER2, basal-like carcinoma (BLC), and unclassified triple-negative carcinoma (UTNC) by established surrogate immunohistochemical profiles. GCDFP and MGB were less likely to be expressed in BLC than in HER2 cancers (P = .000021 and P = .013, respectively) or luminal cancers (P = .00002 and P = .00008, respectively). However, the difference in GCDFP or MGB expression between HER2 and luminal cancers was not significant (P = 1.0 and P = .671, respectively). Our results suggest that luminal cancers demonstrate similar degrees of apocrine differentiation as HER2 cancers. Most BLCs and UTNCs are negative for MGB and GCDFP. Correlation with clinical findings may be needed to exclude the possibility of a metastasis to the breast when BLCs or UTNCs are encountered in a limited sample such as a core biopsy sample.
Topics: Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Basal Cell; Carcinoma, Ductal, Breast; Carrier Proteins; Female; Glycoproteins; Humans; Mammaglobin A; Membrane Transport Proteins; Middle Aged; Neoplasm Proteins; Receptor, ErbB-2; Tissue Array Analysis; Uteroglobin
PubMed: 21411781
DOI: 10.1309/AJCPMFR6OA8ICHNH -
Cancer Research Feb 2011Pathologic axillary lymph node (ALN) status is an important prognostic factor for staging breast cancer. Currently, status is determined by histopathology following...
Pathologic axillary lymph node (ALN) status is an important prognostic factor for staging breast cancer. Currently, status is determined by histopathology following surgical excision of sentinel lymph node(s), which is an invasive, time consuming, and costly procedure with potential morbidity to the patient. Here, we describe an imaging platform for noninvasive assessment of ALN status, eliminating the need for surgical examination of patients to rule out nodal involvement. A targeted imaging probe (MamAb-680) was developed by conjugation of a mammaglobin-A-specific monoclonal antibody to a near-infrared fluorescent dye. Using DNA and tissue microarray, mammaglobin-A was validated as a cell-surface target that is expressed in ALN-positive patient samples but is not expressed in normal lymph nodes. In vivo selectivity was determined by i.v. injection of MamAb-680 into mice with mammaglobin-A-positive and -negative mammary fat pad (MFP) tumors; and by peritumoral MFP injection of the targeted imaging probe in mice with spontaneous ALN metastases. Fluorescence imaging showed that probe was only retained in positive tumors and metastases. As few as 1,000 cells that endogenously express mammaglobin-A were detected in ALN, indicating high sensitivity of this method. Translation of this approach offers considerable potential as a noninvasive clinical strategy to stage breast cancer.
Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Breast Neoplasms; Diagnostic Imaging; Female; Fluorescent Dyes; Humans; Immunoconjugates; Lymph Nodes; Lymphatic Metastasis; Mammaglobin A; Mice; Mice, Nude; Neoplasm Proteins; RNA, Messenger; Transplantation, Heterologous; Uteroglobin
PubMed: 21169406
DOI: 10.1158/0008-5472.CAN-10-3091 -
Cancer Science Nov 2010( 2010; 101: 2501)
( 2010; 101: 2501)
Topics: Biomarkers, Tumor; Breast; Breast Neoplasms; Female; Humans; Lymphatic Metastasis; Mammaglobin A; Neoplasm Proteins; Reverse Transcriptase Polymerase Chain Reaction; Sentinel Lymph Node Biopsy; Uteroglobin
PubMed: 20950374
DOI: 10.1111/j.1349-7006.2010.01715.x -
The International Journal of Biological... 2010To investigate the diagnostic, predictive, and prognostic value of the detection of circulating tumor cells (CTCs) using a three-marker (CK19, hMAM and CEA) RT-PCR assay...
Detection of cytokeratin 19, human mammaglobin, and carcinoembryonic antigen-positive circulating tumor cells by three-marker reverse transcription-PCR assay and its relation to clinical outcome in early breast cancer.
AIMS
To investigate the diagnostic, predictive, and prognostic value of the detection of circulating tumor cells (CTCs) using a three-marker (CK19, hMAM and CEA) RT-PCR assay in patients with early breast cancer.
PATIENTS AND METHODS
Peripheral blood was obtained from 50 patients with early-stage breast cancer before any systemic adjuvant therapy and analyzed for the presence of CK-19, hMAM and CEA mRNA-positive CTCs using an RT-PCR assay. The specificity of the primers used was evaluated in 20 healthy individuals, 24 patients with benign breast disease, and 30 patients with metastatic breast cancer. The detection of CTCs was correlated with clinical outcome.
RESULTS
The detection rate of three-marker-positive CTCs in the blood of patients with early breast cancer was 54.0%, significantly higher than in patients with benign breast disease and healthy blood donors (p=0.002 and p=0.000, respectively). The three-marker RT-PCR assay had 58.8% sensitivity in the parallel test and 100% specificity for CTC detection in the serial test, which was higher than the sensitivity and specificity of single-marker assays. For early breast cancer, correlation analysis between detection of three-marker-positive CTCs and clinicopathological characteristics indicated that detection of threemarker-positive CTCs was significantly correlated with elevated serum CEA levels (p=0.001). After three years of follow-up, 13 of the 27 patients with three-marker-positive CTCs in their blood had relapsed and detection of three-marker-positive CTCs was significantly associated with locoregional recurrence and/or distant metastasis (p=0.002). Detection of three-marker-positive CTCs in peripheral blood was an independent risk factor for reduced median relapse-free interval (p=0.000).
CONCLUSION
The three-marker RT-PCR assay can enhance the sensitivity and specificity of CTC detection compared to singlemarker assay. Detection of three-marker-positive CTCs was associated with relapse and might have important predictive and prognostic implications in early breast cancer.
Topics: Adult; Aged; Biomarkers, Tumor; Breast Neoplasms; Carcinoembryonic Antigen; Carcinoma; Cell Line, Tumor; Disease Progression; Early Detection of Cancer; Female; Humans; Keratin-19; Mammaglobin A; Middle Aged; Neoplasm Proteins; Neoplastic Cells, Circulating; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; Sensitivity and Specificity; Uteroglobin
PubMed: 20586026
DOI: 10.1177/172460081002500201 -
Breast Cancer Research and Treatment May 2011Mammaglobin-A (MGBA), a 10-kD protein, is over expressed in 80% of primary and metastatic human breast cancers. Breast cancer patients demonstrate high frequencies of...
Mammaglobin-A (MGBA), a 10-kD protein, is over expressed in 80% of primary and metastatic human breast cancers. Breast cancer patients demonstrate high frequencies of CD8(+) cytotoxic T lymphocytes (CTL) specific to MGBA. Defining CD8(+) CTL responses to HLA class I-restricted MGBA-derived epitopes assumes significance in the context of our ongoing efforts to clinically translate vaccine strategies targeting MGBA for prevention and/or treatment of human breast cancers. In this study, we define the CD8(+) CTL response to MGBA-derived candidate epitopes presented in the context of HLA-B7, which has a frequency of 17.7% in Caucasian and 15.5% in African American populations. We identified seven MGBA-derived candidate epitopes with high predicted binding scores for HLA-B7 using a computer algorithm. Membrane stabilization studies with TAP-deficient T2 cells transfected with HLA-B7 indicated that MGBA B7.3 (VSKTEYKEL), B7.6 (KLLMVLMLA), B7.7 (NPQVSKTEY), and B7.1 (YAGSGCPLL) have the highest HLA-B7 binding affinities. Further, two CD8(+) CTL cell lines generated in vitro against T2.B7 cells individually loaded with MGBA-derived candidate epitopes showed significant cytotoxic activity against MGBA B7.1, B7.3, B7.6, and B7.7. In addition, the same CD8(+) CTL lines lysed the HLA-B7(+)/MGBA(+) human breast cancer cell line DU-4475 but had no significant cytotoxicity against HLA-B7(-) or MGBA(-) breast cancer cell lines. Cold-target inhibition studies strongly suggest that MGBA B7.3 is an immunodominant epitope. In summary, our results define HLA-B7-restriced, MGBA-derived, CD8(+) CTL epitopes with all of the necessary features for developing novel vaccine strategies against HLA-B7 expressing breast cancer patients.
Topics: Antibodies, Monoclonal; Antibody Affinity; Antibody Specificity; Breast Neoplasms; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cytotoxicity, Immunologic; Epitopes, T-Lymphocyte; Female; HLA-B7 Antigen; Humans; Mammaglobin A; Neoplasm Proteins; T-Lymphocytes, Cytotoxic; Uteroglobin
PubMed: 20544273
DOI: 10.1007/s10549-010-0975-z -
Breast (Edinburgh, Scotland) Oct 2010There are limited data that compare the utility of immunohistochemical detection of mammaglobin with Gross Cystic Disease Fluid Protein-15 (GCDFP-15) in locally...
There are limited data that compare the utility of immunohistochemical detection of mammaglobin with Gross Cystic Disease Fluid Protein-15 (GCDFP-15) in locally recurrent and metastatic breast cancers. Forty-three local and 72 distant recurrences of breast cancer, 8 metastatic lesions to the breast from other organs, and 30 metastases from non-breast primaries were immunohistochemically stained with mammaglobin and GCDFP-15 antibodies. Mammaglobin was expressed in 55 (47.8%) and GCDFP-15 detected in 13 (11.3%) locally and distantly recurrent breast cancers. A higher percentage of tumor cells was stained with mammaglobin at greater staining intensity than GCDFP-15, for both metastatic and locally recurrent breast cancers. The difference in staining intensity as well as mean percentage of tumor cells stained for both markers was statistically significant (p < 0.005). Metastases to the breast from other organs and metastatic lesions from non-breast primaries were uniformly negative for both mammaglobin and GCDFP-15. Our study demonstrates that immunohistochemical analysis of mammaglobin is superior to GCDFP-15 in detecting a tumor of breast origin, and can be incorporated into immunohistochemical panels evaluating tumors from unknown primary sites.
Topics: Adenocarcinoma, Mucinous; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Neoplasms; Carcinoma, Ductal, Breast; Carcinoma, Lobular; Carcinoma, Papillary; Carrier Proteins; Female; Glycoproteins; Humans; Immunohistochemistry; Mammaglobin A; Membrane Transport Proteins; Middle Aged; Neoplasm Metastasis; Neoplasm Proteins; Neoplasm Recurrence, Local; Neoplasms, Unknown Primary; Sensitivity and Specificity; Uteroglobin
PubMed: 20347310
DOI: 10.1016/j.breast.2010.02.007 -
Vaccine Feb 2010It is commonly believed that delivery of antigen into the class I antigen presentation pathway is a limiting factor in the clinical translation of DNA vaccines. This is...
It is commonly believed that delivery of antigen into the class I antigen presentation pathway is a limiting factor in the clinical translation of DNA vaccines. This is of particular concern in the context of cancer vaccine development as many immunodominant peptides derived from self tumor antigens are not processed and presented efficiently. To address this limitation, we have engineered completely assembled peptide/MHC class I complexes whereby all three components (class I heavy chain, beta(2)m, and peptide) are attached by flexible linkers and expressed as a single polypeptide (single chain trimers or SCT). In this study, we tested the efficacy of progressive generations of SCT DNA vaccines engineered to (1) enhance peptide binding, (2) enhance interaction with the CD8 coreceptor, and/or (3) activate CD4(+) helper T cells. Disulfide trap SCT (dtSCT) have been engineered to improve peptide binding, with mutations designed to create a disulfide bond between the class I heavy chain and the peptide linker. dtSCT DNA vaccines dramatically enhance the immune response to model low affinity antigens as measured by ELISPOT analysis and tumor challenge. SCT engineered to enhance interaction with the CD8 coreceptor have a higher affinity for the TCR/CD8 complex, and are associated with more robust CD8(+) T cell responses following vaccination. Finally, SCT constructs that coexpress a universal helper epitope PADRE, dramatically enhance CD8(+) T cell responses. Taken together, our data demonstrate that dtSCT DNA vaccines coexpressing a universal CD4 epitope are highly effective in generating immune responses to poorly processed and presented cancer antigens.
Topics: Animals; Antigen Presentation; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; CHO Cells; Cell Line, Tumor; Cricetinae; Cricetulus; Epitopes, T-Lymphocyte; Genes, MHC Class I; HLA-A2 Antigen; Humans; Lymphocyte Activation; Mammaglobin A; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mutagenesis, Site-Directed; Neoplasm Proteins; Ovalbumin; Protein Binding; Protein Engineering; Uteroglobin; Vaccines, DNA; beta 2-Microglobulin
PubMed: 20188246
DOI: 10.1016/j.vaccine.2009.10.096 -
Genetics and Molecular Research : GMR 2010Detection of residual tumor cells in the circulation can provide prognostic as well as therapeutic information and help in identifying patients at high risk for...
Detection of residual tumor cells in the circulation can provide prognostic as well as therapeutic information and help in identifying patients at high risk for developing metastases. Maspin and mammaglobin are two molecules that are specifically associated with breast cancer. We looked for mammaglobin and maspin transcripts in the peripheral blood of patients with breast cancer and evaluated their utility as a marker of the response to therapy. Maspin and mammaglobin transcripts were analyzed in 85 breast-cancer patients by nested RT-PCR, prior to and after treatment. Before therapy, 10 patients were found positive for mammaglobin and 20 patients were positive for maspin. In four patients, both transcripts were detected. Immediately following treatment, only one patient was still positive for mammaglobin while maspin transcripts persisted in three patients. Disease progression was observed mainly in patients in whom maspin transcripts were not detectable. Molecular detection of circulating tumor cells during therapy based on analysis for mammaglobin and maspin transcripts is an easy and practical method that can be applied to follow-up patients. We suggest that detection of mammaglobin mRNA is useful to determine the effect of therapy while maspin transcripts may indicate more aggressive disease.
Topics: Adult; Biomarkers, Tumor; Breast Neoplasms; Disease Progression; Female; Humans; Mammaglobin A; Middle Aged; Neoplasm Proteins; Neoplastic Cells, Circulating; Prognosis; RNA, Messenger; RNA, Neoplasm; Serpins; Transcription, Genetic; Uteroglobin
PubMed: 20092039
DOI: 10.4238/vol9-1gmr649 -
European Journal of Medical Research Sep 2009The diagnostic tools to predict the prognosis in patients suffering from breast cancer (BC) need further improvements. New technological achievements like the gene...
BACKGROUND
The diagnostic tools to predict the prognosis in patients suffering from breast cancer (BC) need further improvements. New technological achievements like the gene profiling of circulating tumour cells (CTC) could help identify new prognostic markers in the clinical setting. Furthermore, gene expression patterns of CTC might provide important informations on the mechanisms of tumour cell metastasation.
MATERIALS AND METHODS
We performed realtime-PCR and multiplex-PCR analyses following immunomagnetic separation of CTC. Peripheral blood (PB) samples of 63 patients with breast cancer of various stages were analyzed and compared to a control group of 14 healthy individuals. After reverse-transcription, we performed multiplex PCR using primers for the genes ga733.3, muc-1 and c-erbB2. Mammaglobin1, spdef and c-erbB2 were analyzed applying realtime-PCR.
RESULTS
ga733.2 overexpression was found in 12.7% of breast cancer cases, muc-1 in 15.9%, mgb1 in 9.1% and spdef in 12.1%. In this study, c-erbB2 did not show any significant correlation to BC, possibly due to a highly ambient expression. Besides single gene analyses, gene profiles were additionally evaluated. Highly significant correlations to BC were found in single gene analyses of ga733.2 and muc-1 and in gene profile analyses of ga733.3*muc-1 and GA7 ga733.3*muc-1*mgb1*spdef.
CONCLUSION
Our study reveals that the single genes ga733.3, muc-1 and the gene profiles ga733.3*muc-1 and ga733.3*3muc-1*mgb1*spdef can serve as markers for the detection of CTC in BC. The multigene analyses found highly positive levels in BC patients. Our study indicates that not single gene analyses but subtle patterns of multiple genes lead to rising accuracy and low loss of specificity in detection of breast cancer cases.
Topics: Adult; Aged; Breast Neoplasms; Female; Gene Expression Profiling; Humans; Immunohistochemistry; Mammaglobin A; Middle Aged; Mucin-1; Neoplasm Proteins; Neoplastic Cells, Circulating; Polymerase Chain Reaction; Receptor, ErbB-2; Uteroglobin
PubMed: 19748849
DOI: 10.1186/2047-783x-14-10-426 -
Romanian Journal of Morphology and... 2009Mammaglobin A is a specific marker of the normal and neoplastic mammary tissue that usually is detected by RT-PCR. Few data are available about the immunohistochemical...
Mammaglobin A is a specific marker of the normal and neoplastic mammary tissue that usually is detected by RT-PCR. Few data are available about the immunohistochemical expression of this marker in mammary carcinoma and about the significance of the positive reaction. Our purpose was to investigate the sensitivity of the mammaglobin expression in breast cancer and to determine its correlations with conventional prognostic parameters. There were investigated 47 patients with breast carcinoma, and slides from paraffin blocks were stained with an antibody against mammaglobin. The immunohistochemical reaction was scored based on the percentage of positive tumor cells in both primary tumors and lymph node metastasis. Positive reaction for mammaglobin was found in the normal mammary tissue adjacent to the tumor in all cases, in 78.72% primary breast carcinoma, and in 58.06% of cases with lymph node metastases. A significant correlation was found between the mammaglobin expression in the primary tumor, grade, and lymph node status, but not with the age of the patient, pathologic subtype of carcinoma and stage of the tumor. The ductal in situ carcinoma associated to the invasive tumor did not influence significantly the prognostic value of mammaglobin expression. Out results suggest that mammaglobin is a sensitive marker of breast carcinoma, it defines a subgroup of patients with better prognosis and is a useful method to detect breast cancer metastases.
Topics: Adult; Aged; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Female; Humans; Immunohistochemistry; Lymphatic Metastasis; Mammaglobin A; Middle Aged; Neoplasm Proteins; Tumor Cells, Cultured; Uteroglobin
PubMed: 19690758
DOI: No ID Found