Apocrine Cysts of the Breast Biomarkers, Origin, Enlargement, and Relation with Cancer Phenotype

Danish Centre for Translational Breast Cancer Research (DCTB), Copenhagen, Denmark.
Molecular & Cellular Proteomics (Impact Factor: 6.56). 04/2006; 5(3):462-83. DOI: 10.1074/mcp.M500348-MCP200
Source: PubMed


Up to one-third of women aged 30-50 years have cysts in their breasts and are presumed to be at increased risk of developing breast cancer. Here we present an extensive proteomic and immunohistochemistry (IHC) study of breast apocrine cystic lesions aimed at generating specific biomarkers and elucidating the relationship, if existent, of apocrine cysts with cancer phenotype. To this end we compared the expression profiles of apocrine macrocysts obtained from mastectomies from high risk cancer patients with those of cancerous and non-malignant mammary tissue biopsies collected from the same patients. We identified two biomarkers, 15-hydroxyprostaglandin dehydrogenase and 3-hydroxymethylglutaryl-CoA reductase, that were expressed specifically by apocrine type I cysts as well as by apocrine metaplastic cells in type II microcysts, terminal ducts, and intraductal papillary lesions. No expression of these markers was observed in non-malignant terminal ductal lobular units, type II flat cysts, stroma cells, or fat tissue as judged by IHC analysis of matched non-malignant tissue samples collected from 93 high risk patients enrolled in our cancer program. IHC analysis of the corresponding 93 primary tumors indicated that most apocrine changes have little intrinsic malignant potential, although some may progress to invasive apocrine cancer. None of the apocrine lesions examined, however, seemed to be a precursor of invasive ductal carcinomas, which accounted for 81% of the tumors analyzed. Our studies also provided some insight into the origin, development, and enlargement of apocrine cysts in mammary tissue. The successful identification of differentially expressed proteins that characterize specific steps in the progression from early benign lesions to apocrine cancer opens a window of opportunity for designing and testing new approaches for pharmacological intervention, not only in a therapeutic setting but also for chemoprevention, to inhibit cyst development as both 15-hydroxyprostaglandin dehydrogenase and 3-hydroxymethylglutaryl-CoA reductase are currently being targeted for chemoprevention strategies in various malignancies.

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Available from: Julio Celis, Jan 05, 2016
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    • "Fibrocystic changes are the most frequent benign disorder of the breast (Guray et al., 2006). Cysts contains a variety of biologically active substances including steroids, cytokines, growth factors as well as proteins (Celis et al., 2006). Previous workers found that expression of HGF and its receptor c -Met is correlated with breast carcinoma progression (Beviglia et al., 1999). "
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    ABSTRACT: Hepatocyte growth factor (HGF) also known as scatter factor (SF) and its receptor c-met play important roles in mammary differentiation and have been implicated in mammary carcinogenesis. Objective: Estimation of the plasma level of HGF in females with benign breast lumps or breast carcinomas and correlating levels with important prognostic parameters. Subjects: Sixty eight adult premenopausal females were divided into control group of fifteen healthy volunteers and fifty-three patients subdivided into fifteen with benign breast lumps and thirty-eight with breast carcinomas. A thorough clinical examination, plain chest x-rays, ultrasonography of the abdomen and pelvis, pre- operative fine needle aspiration cytology, estimation of fasting serum glucose, urea, creatinine and uric acid levels, alanine aminotransferase activities, C-reactive protein, HGF level and histopathological examination of the breast masses were performed. Significant increase in HGF levels was found in patients with benign breast lumps and in breast cancer patients when each was compared to controls and when cancer patients were compared to the benign breast lumps group. The serum level of HGF is an independent prognostic indicator for breast cancer.
    Preview · Article · Jan 2010 · Asian Pacific journal of cancer prevention: APJCP
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    • "By comparing the gel-based protein expression profiles of ''blue dome'' apocrine cysts with normal breast epithelial tissue obtained from the same patient (Celis et al., 2006a,b, 2007a,b, 2008), we have identified a number of apocrine protein biomarkers and two of these, 15-PGDH (Figure 1A) and ACSM1 (Figure 1B), were proven to be specific for benign apocrine lesions as determined by IHC and 2D gel-based proteomics (Celis et al., 2006a,b, 2007a,b, 2008). IHC analysis of well-defined sets of breast tumour subtypes without apocrine differentiation, has failed to show any reactivity with the 15- PGDH and ACSM1 antibodies at the dilutions used in this study (Celis et al., 2008). "
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    ABSTRACT: Invasive apocrine carcinomas (IACs), as defined by morphological features, correspond to 0.3-4% of all invasive ductal carcinomas (IDC), and despite the fact that they are histologically distinct from other breast lesions there are currently no standard molecular criteria available for their diagnosis and no unequivocal information as to their prognosis. In an effort to address these concerns we have been using protein expression profiling technologies in combination with mass spectrometry and immunohistochemistry (IHC) to discover specific biomarkers that could allow us to molecularly characterize these lesions as well as to dissect some of the steps in the processes underlying breast apocrine metaplasia and development of precancerous apocrine lesions. Establishing these apocrine-specific markers as best practice for the routine pathology evaluation of breast cancer, however, will require their validation in large cohorts of patients. Towards this goal we have composed a panel of antibodies against components of an apocrine protein signature that includes probes against the apocrine-specific markers 15-prostaglandin dehydrogenase (15-PGDH), and acyl-CoA synthetase medium-chain family member 1 (ACSM1), in addition to a set of categorizing markers that are consistently expressed (AR, CD24) or not expressed (ERα, PgR, Bcl-2, and GATA-3) by apocrine metaplasia in benign breast lesions and apocrine sweat glands. This panel was used to analyze a well-defined cohort consisting of 14 apocrine ductal carcinoma in situ (ADCIS), and 33 IACs diagnosed at the Cancer Institute Hospital, Tokyo between 1997 and 2001. Samples were originally classified on the basis of cellular morphology with all cases having more than 90% of the tumour cells exhibiting cytological features typical of apocrine cells. Using the expression of 15-PGDH and/or ACSM1 as the main criterion, but taking into account the expression of other markers, we were able to identify unambiguously 13 out of 14 ADCIS (92.9%) and 20 out of 33 (60.6%) IAC samples, respectively, as being of apocrine origin. Our results demonstrate that IACs correspond to a distinct, even if heterogeneous, molecular subgroup of breast carcinomas that can be readily identified in an unbiased way using a combination of markers that recapitulate the phenotype of apocrine sweat glands (15-PGDH(+), ACSM1(+), AR(+), CD24(+), ERα(-), PgR(-), Bcl-2(-), and GATA-3(-)). These results pave the way for addressing issues such as prognosis of IACs, patient stratification for targeted therapeutics, as well as research strategies for identifying novel therapeutic targets for developing new cancer therapies.
    Full-text · Article · Mar 2009 · Molecular oncology
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    • "A variety of experiments suggest a role of cholesterol in the biology of PGRMC1, as reviewed by Cahill [20]. The rate-limiting enzyme of the mevalonate pathway leading to cholesterol synthesis is hydroxymethylglutarate-coenzyme A reductase, and this enzyme is both regulated by cholesterol levels [36,37] and is diagnostic of a recently identified class of poor prognosis apocrine breast cancers that were both ER-α and progesterone receptor negative [38]. "
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    ABSTRACT: Breast tumors lacking the estrogen receptor-alpha (ER-alpha) have increased incidence of resistance to therapy and poorer clinical prognosis. Whole tissue sections from 16 cryopreserved breast cancer tumors that were either positive or negative for the ER (eight ER positive and eight ER negative) were differentially analyzed by multiplex imaging of two-dimensional PAGE gels using 54 cm isoelectric focusing. Differentially detected spots of Progesterone Receptor Membrane Component 1 (PGRMC1) were shown to differ in phosphorylation status by differential two dimensional polyacrylamide gel electrophoresis of phosphatase-treated tumor proteins. Site directed mutagenesis was used to create putative phosphorylation site point mutants in PGRMC1. Stable transfectants of these mutants in MCF7 cells were assayed for their survival after oxidative stress, and for AKT kinase phosphorylation. Immune fluorescence using anti-PGRMC1 monoclonal antibody 5G7 was performed on breast cancer tissue microarrays. Proteins significantly differentially abundant between estrogen receptor negative and estrogen receptor positive tumors at the 0.1% level were consistent with published profiles, suggesting an altered keratin pool, and increased inflammation and wound responses in estrogen receptor negative tumors. Two of three spots of PGRMC1 were more abundant in estrogen receptor negative tumors. Phosphatase treatment of breast tumor proteins indicated that the PGRMC1 isoforms differed in their phosphorylation status. Simultaneous mutation of PGRMC1 serine-56 and serine-180 [corrected] fully abrogated the sensitivity of stably transfected MCF7 breast cancer cells to peroxide-induced cell death. Immune fluorescence revealed that PGRMC1 was primarily expressed in ER-negative basal epithelial cells of mammary ductules. Even in advanced tumors, high levels of ER or PGRMC1 were almost mutually exclusive in individual cells. In five out of five examined ductal in situ breast cancers of comedo type, PGRMC1 was expressed in glucose transporter 1 negative or positive poorly oxygenated cells surrounding the necrotic core, surrounded by a more distal halo of ER-positive cells. PGRMC1 phosphorylation may be involved in the clinical differences that underpin breast tumors of differing ER status.
    Full-text · Article · Nov 2008 · Breast cancer research: BCR
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