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Cancer Genomics & Proteomics 2022Until now, little emphasis has been placed on the protein expression profile of male breast cancer (MBC) tumors, due to the rarity of the disease. The present study...
BACKGROUND/AIM
Until now, little emphasis has been placed on the protein expression profile of male breast cancer (MBC) tumors, due to the rarity of the disease. The present study aimed to identify a proteomic pattern that is characteristic for malignant male breast tissue epithelium.
MATERIALS AND METHODS
The protein content of four male breast tumors and corresponding adjacent healthy (control) tissues was analyzed by high-throughput nano-liquid chromatography-MS/MS technology.
RESULTS
A total of 2,352 proteins were identified, that correspond to 1,249 single gene products, with diverse biological roles. Of those, a panel of 119 differentially expressed tissue proteins was identified in MBC samples compared to controls; 90 were found to be over-expressed in MBC tissues, while 29 were down-regulated. Concurrently, 844 proteins were detected only in MBC tumors and 197 were expressed exclusively in control mammary samples.
CONCLUSION
Differential proteomic expression was found in MBC tissue, leading to improved understanding of MBC pathology and highlighting the need for personalized management of male patients.
Topics: Biomarkers, Tumor; Breast; Breast Neoplasms; Breast Neoplasms, Male; Chromatography, Liquid; Humans; Male; Proteomics; Tandem Mass Spectrometry
PubMed: 35181590
DOI: 10.21873/cgp.20316 -
PeerJ 2023The epithelial-mesenchymal transition (EMT) is a multi-step morphogenetic process in which epithelial cells lose their epithelial properties and gain mesenchymal...
BACKGROUND
The epithelial-mesenchymal transition (EMT) is a multi-step morphogenetic process in which epithelial cells lose their epithelial properties and gain mesenchymal characteristics. The process of EMT has been shown to mediate mammary gland fibrosis. Understanding how mesenchymal cells emerge from an epithelial default state will aid in unravelling the mechanisms that control fibrosis and, ultimately, in identifying therapeutic targets to alleviate fibrosis.
METHODS
The effects of EGF and high glucose (HG) on EMT in mammary epithelial cells, MCF10A and GMECs, as well as their pathogenic role, were studied. analysis was used to find interacting partners and protein-chemical/drug molecule interactions.
RESULTS
On treatment with EGF and/or HG, qPCR analysis showed a significant increase in the gene expression of EMT markers and downstream signalling genes. The expression of these genes was reduced on treatment with EGF+HG combination in both cell lines. The protein expression of COL1A1 increased as compared to the control in cells treated with EGF or HG alone, but when the cells were treated with EGF and HG together, the protein expression of COL1A1 decreased. ROS levels and cell death increased in cells treated with EGF and HG alone, whereas cells treated with EGF and HG together showed a decrease in ROS production and apoptosis. analysis of protein-protein interactions suggest the possible role of MAPK1, actin alpha 2 (ACTA2), COL1A1, and NFB1 in regulating TGF1, ubiquitin C (UBC), specificity protein 1 (SP1) and E1A binding protein P300 (EP300). Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment suggests advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signalling pathway, relaxin signalling pathway and extra cellular matrix (ECM) receptor interactions underlying fibrosis mechanism.
CONCLUSION
This study demonstrates that EGF and HG induce EMT in mammary epithelial cells and may also have a role in fibrosis.
Topics: Humans; Epithelial-Mesenchymal Transition; Epidermal Growth Factor; Mammary Glands, Human; Reactive Oxygen Species; Fibrosis
PubMed: 37187521
DOI: 10.7717/peerj.15207 -
IUBMB Life May 2017Epithelial cell layers are interconnected by a meshwork of tight junction (TJ) protein strands, which are localized within apicolateral membranes. The proteins that form... (Review)
Review
Epithelial cell layers are interconnected by a meshwork of tight junction (TJ) protein strands, which are localized within apicolateral membranes. The proteins that form TJs are regarded to provide a static barrier, determining epithelial properties. However, recent findings in the field of barriology suggest that TJs contribute to more physiological aspects than indicated by the sum of the qualities of the single TJ proteins. Generally, TJs exhibit four major functions: (i) a "gate function," defining transepithelial permeability (i.e., barrier) properties, (ii) a "fence function" determining epithelial cell polarity, (iii) a "signaling function," affecting regulatory pathways, and (iv) a "stabilizing function," maintaining the integrity of the epithelium. This review presents a critical view on how the efficacy of physiological processes in epithelia and thus organ function might be improved by changes in the expression of claudins, the latter representing the largest and most variable family of TJ proteins. Major focus is set on (i) the coordinated regulation of transport and barrier in the intestine, (ii) the role of TJs in defining the route for antigen uptake and presentation in intestinal Peyer's patches, and (iii) the TJ function in mammary glands in response to milk accumulation, which represent impressive examples to highlight the amplification of epithelial functions by TJ proteins. © 2017 IUBMB Life, 69(5):290-296, 2017.
Topics: Animals; Claudins; Epithelial Cells; Epithelium; Humans; Intestinal Mucosa; Mammary Glands, Human; Permeability; Sodium; Tight Junctions
PubMed: 28371008
DOI: 10.1002/iub.1622 -
Journal of Mammary Gland Biology and... Oct 2004The structure and function of each individual mammary epithelial cell (MEC) is largely controlled by a bidirectional interchange of chemical and mechanical signals with... (Review)
Review
The structure and function of each individual mammary epithelial cell (MEC) is largely controlled by a bidirectional interchange of chemical and mechanical signals with the microenvironment. Most of these signals are tissue-specific, since they arise from the three-dimensional (3D) tissue organization and are modulated during mammary gland development, maturation, pregnancy, lactation, and involution. Although the important role played by structural and mechanical signals in mammary cell and tissue function is being increasingly recognized, quantitative biomechanical approaches are still scarce. Here we review currently available biomechanical tools that allow quantitative examination of individual cells, groups of cells or full monolayers in two-dimensional cultures, and cells in 3D cultures. Current technological limitations and challenges are discussed, with special emphasis on their potential applications in MEC biology. We argue that the combination of biomechanical tools with current efforts in mathematical modeling and in cell and molecular biology applied to 3D cultures provides a powerful approach to unravel the complexity of tissue-specific structure-function relationships.
Topics: Animals; Biology; Biomechanical Phenomena; Cell Physiological Phenomena; Epithelium; Humans; Mammary Glands, Human
PubMed: 15838605
DOI: 10.1007/s10911-004-1406-8 -
Cold Spring Harbor Protocols Dec 2015Cleared mammary fat pad (MFP) transplantation has been a standard technique for studies of mammary development and cancer for several decades. The mammary gland is...
Cleared mammary fat pad (MFP) transplantation has been a standard technique for studies of mammary development and cancer for several decades. The mammary gland is comprised of several fundamental components: The epithelial compartment contains basal/myoepithelial cells and luminal cells, and the stromal compartment (called the MFP) contains adipocytes, smooth muscle cells, fibroblasts, and immune cells. In 3- to 4-wk-old female mice, the mammary epithelium is concentrated very close to the nipple and has not yet grown beyond the mammary lymph node to penetrate the bulk of the MFP. This developmental feature provides an anatomical fixed point, and enables one to cut away the portion of the MFP from the nipple to the lymph node, leaving behind the majority of the MFP free of epithelium. The "cleared" MFP can serve as a supportive native microenvironment fully sufficient for the organogenesis of injected donor epithelium. Normal mammary epithelial donor cells will produce histologically and functionally normal mammary ductal epithelium several weeks posttransplant, with the exception that the ducts will not be connected to the nipple. The assay described here provides a powerful platform for assessing the developmental and tumorigenic potential of engineered cells of interest.
Topics: Adipose Tissue; Animals; Carcinogenesis; Epithelial Cells; Humans; Mammary Glands, Human; Mice; Models, Animal; Organogenesis; Transplantation
PubMed: 26631119
DOI: 10.1101/pdb.prot078071 -
EMBO Molecular Medicine Feb 2020During obesity, macrophages infiltrate the breast tissue leading to low-grade chronic inflammation, a factor considered responsible for the higher risk of breast cancer...
During obesity, macrophages infiltrate the breast tissue leading to low-grade chronic inflammation, a factor considered responsible for the higher risk of breast cancer associated with obesity. Here, we formally demonstrate that breast epithelial cells acquire malignant properties when exposed to medium conditioned by macrophages derived from human healthy donors. These effects were mediated by the breast cancer oncogene IKKε and its downstream target-the serine biosynthesis pathway as demonstrated by genetic or pharmacological tools. Furthermore, amlexanox, an FDA-approved drug targeting IKKε and its homologue TBK1, delayed in vivo tumour formation in a combined genetic mouse model of breast cancer and high-fat diet-induced obesity/inflammation. Finally, in human breast cancer tissues, we validated the link between inflammation-IKKε and alteration of cellular metabolism. Altogether, we identified a pathway connecting obesity-driven inflammation to breast cancer and a potential therapeutic strategy to reduce the risk of breast cancer associated with obesity.
Topics: Aminopyridines; Animals; Breast Neoplasms; Culture Media, Conditioned; Epithelial Cells; Female; Humans; I-kappa B Kinase; Inflammation; Macrophages; Mammary Glands, Human; Mice; Obesity; Protein Serine-Threonine Kinases; Serine
PubMed: 31930708
DOI: 10.15252/emmm.201910491 -
The American Journal of Pathology Jan 1999Tcf-4 is a member of the Tcf/Lef family of transcription factors that interact functionally with beta-catenin to mediate Wnt signaling in vertebrates. We have previously...
Tcf-4 is a member of the Tcf/Lef family of transcription factors that interact functionally with beta-catenin to mediate Wnt signaling in vertebrates. We have previously demonstrated that the tumor suppressor function of APC in the small intestine is mediated via regulation of Tcf-4/beta-catenin transcriptional activity. To gain further insight into the role of Tcf-4 in development and carcinogenesis we have generated several mouse monoclonal antibodies, one of which is specific for Tcf-4 and another of which recognizes both Tcf-3 and Tcf-4. Immunohistochemistry performed with the Tcf 4- specific monoclonal antibody revealed high levels of expression in normal intestinal and mammary epithelium and carcinomas derived therefrom. Additional sites of Tcf-3 expression, as revealed by staining with the Tcf-3/-4 antibody, occurred only within the stomach epithelium, hair follicles, and keratinocytes of the skin. A temporal Tcf-4 expression gradient was observed along the crypt-villus axis of human small intestinal epithelium: strong Tcf-4 expression was present within the crypts of early (week 16) human fetal small intestine, with the villi showing barely detectable Tcf-4 protein levels. Tcf-4 expression levels increased dramatically on the villi of more highly developed (week 22) fetal small intestine. We conclude that Tcf-4 exhibits a highly restricted expression pattern related to the developmental stage of the intestinal epithelium. The high levels of Tcf-4 expression in mammary epithelium and mammary carcinomas may also indicate a role in the development of this tissue and breast carcinoma.
Topics: Animals; Antibodies, Monoclonal; Breast; Breast Neoplasms; COS Cells; Carcinoma; Cell Nucleus; Colonic Neoplasms; Cytoskeletal Proteins; Epithelium; Female; Fetus; HMGB Proteins; Humans; Immunohistochemistry; Intestinal Mucosa; Intestines; Mice; Mice, Inbred BALB C; Staining and Labeling; TCF Transcription Factors; Trans-Activators; Transcription Factor 7-Like 1 Protein; Transcription Factor 7-Like 2 Protein; Transcription Factors; beta Catenin
PubMed: 9916915
DOI: 10.1016/S0002-9440(10)65247-9 -
The Journal of Clinical Investigation Nov 2020Women with dense breasts have an increased lifetime risk of malignancy that has been attributed to a higher epithelial density. Quantitative proteomics, collagen... (Randomized Controlled Trial)
Randomized Controlled Trial
Women with dense breasts have an increased lifetime risk of malignancy that has been attributed to a higher epithelial density. Quantitative proteomics, collagen analysis, and mechanical measurements in normal tissue revealed that stroma in the high-density breast contains more oriented, fibrillar collagen that is stiffer and correlates with higher epithelial cell density. microRNA (miR) profiling of breast tissue identified miR-203 as a matrix stiffness-repressed transcript that is downregulated by collagen density and reduced in the breast epithelium of women with high mammographic density. Culture studies demonstrated that ZNF217 mediates a matrix stiffness- and collagen density-induced increase in Akt activity and mammary epithelial cell proliferation. Manipulation of the epithelium in a mouse model of mammographic density supported a causal relationship between stromal stiffness, reduced miR-203, higher levels of the murine homolog Zfp217, and increased Akt activity and mammary epithelial proliferation. ZNF217 was also increased in the normal breast epithelium of women with high mammographic density, correlated positively with epithelial proliferation and density, and inversely with miR-203. The findings identify ZNF217 as a potential target toward which preexisting therapies, such as the Akt inhibitor triciribine, could be used as a chemopreventive agent to reduce cancer risk in women with high mammographic density.
Topics: Adult; Animals; Breast Neoplasms; Double-Blind Method; Female; Humans; Mammary Glands, Human; Mice; MicroRNAs; Middle Aged; Oncogene Proteins; Proto-Oncogene Proteins c-akt; RNA, Neoplasm; Risk Factors; Trans-Activators
PubMed: 32721948
DOI: 10.1172/JCI129249 -
Oncogene Mar 2018The process of organ development requires a delicate balance between cellular plasticity and differentiation. This balance is disrupted in cancer initiation and...
The process of organ development requires a delicate balance between cellular plasticity and differentiation. This balance is disrupted in cancer initiation and progression. N-Myc and STAT interactor (NMI: human or Nmi: murine) has emerged as a relevant player in the etiology of breast cancer. However, a fundamental understanding of its relevance to normal mammary biology is lacking. To gain insight into its normal function in mammary gland, we generated a mammary-specific Nmi knockout mouse model. We observed that Nmi protein expression is induced in mammary epithelium at the onset of pregnancy, in luminal cells and persists throughout lactation. Nmi knockout results in a precocious alveolar phenotype. These alveoli exhibit an extensive presence of nuclear β-catenin and enhanced Wnt/β-catenin signaling. The Nmi knockout pubertal ductal tree shows enhanced invasion of the mammary fatpad and increased terminal end bud numbers. Tumors from Nmi null mammary epithelium show a significant enrichment of poorly differentiated cells with elevated stem/progenitor markers, active Wnt/β-catenin signaling, highly invasive morphology as well as, increased number of distant metastases. Our study demonstrates that Nmi has a distinct role in the differentiation process of mammary luminal epithelial cell compartment and developmental aberrations resulting from Nmi absence contribute to metastasis and demonstrates that aberration in normal developmental program can lead to metastatic disease, highlighting the contribution and importance of luminal progenitor cells in driving metastatic disease.
Topics: Animals; Breast; Breast Neoplasms; Cell Differentiation; Cells, Cultured; Female; Humans; Intracellular Signaling Peptides and Proteins; Mammary Glands, Animal; Mammary Glands, Human; Mammary Neoplasms, Animal; Mice; Mice, Inbred C57BL; Mice, Knockout; Neoplasm Metastasis; Organogenesis
PubMed: 29326438
DOI: 10.1038/s41388-017-0037-7 -
International Journal of Experimental... Aug 1998The rodent and human nonpregnant mammary glands contain epithelial, intermediate and myoepithelial cells which have all been isolated as cell lines in vitro.... (Review)
Review
The rodent and human nonpregnant mammary glands contain epithelial, intermediate and myoepithelial cells which have all been isolated as cell lines in vitro. Transforming growth factor-alpha (TGF alpha) and basic fibroblast growth factor (bFGF) are produced by myoepithelial cells and can stimulate the growth of intermediate stem cells in vitro. Epithelial and intermediate cells behave like stem cells in vitro, since they can differentiate into alveolar-like an myoepithelial cells. The myoepithelial differentiation pathway is associated with the early expression of a calcium-binding regulatory protein called p9Ka and the protease, Cathepsin D. Myoepithelial cells are also present in benign lesions but not in malignant mammary carcinomas of rats or humans, whose resultant cell lines fail to differentiate completely along the myoepithelial cell pathway. Loss of the myoepithelial cell in some invasive carcinomas may be compensated, at least in part, by changes in malignant cells. Over-expression of TGF alpha and/or erbB receptors may reduce the requirement for TGF alpha, whilst ectopic production of bFGF and its receptors and p9Ka/Cathespin D may assist in tumorigenesis and in metastasis, respectively. Thus compensation for, or retention of, molecules potentially involved in the differentiation of mammary cells may be a mechanism by which malignancy progresses in some human invasive carcinomas.
Topics: Animals; Antigens, Differentiation; Breast; Breast Neoplasms; Cell Differentiation; Cell Transformation, Neoplastic; Epithelium; Female; Growth Substances; Hormones; Humans; Mammary Glands, Animal; Micronutrients; Pituitary Gland; Rats; Stem Cells
PubMed: 9797716
DOI: 10.1046/j.1365-2613.1998.00068.x