scholarly journals Dual recombinase action in the normal and neoplastic mammary gland epithelium

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Patrick D. Rädler ◽  
Kerry Vistisen ◽  
Aleata A. Triplett ◽  
Rayane Dennaoui ◽  
Yong Li ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Cancer Cells ◽  
Mammary Cancer ◽  
Gene Activation ◽  
Metastatic Tumor ◽  
Mammary Tumors ◽  
Lineage Tracing ◽  
Normal Mammary Gland ◽  
Transgenic Mouse Line

AbstractWe developed a transgenic mouse line that expresses the codon-optimized Flp recombinase under the control of the MMTV promoter in luminal epithelial cells of the mammary gland. In this report, we demonstrate the versatile applicability of the new MMTV-Flp strain to manipulate genes in a temporally and spatially controlled manner in the normal mammary gland, in luminal-type mammary tumors that overexpress ERBB2, and in a new KRAS-associated mammary cancer model. Although the MMTV-Flp is expressed in a mosaic pattern in the luminal epithelium, the Flp-mediated activation of a mutant KrasG12D allele resulted in basal-like mammary tumors that progressively acquired mesenchymal features. Besides its applicability as a tool for gene activation and cell lineage tracing to validate the cellular origin of primary and metastatic tumor cells, we employed the MMTV-Flp transgene together with the tamoxifen-inducible Cre recombinase to demonstrate that the combinatorial action of both recombinases can be used to delete or to activate genes in established tumors. In a proof-of-principle experiment, we conditionally deleted the JAK1 tyrosine kinase in KRAS-transformed mammary cancer cells using the dual recombinase approach and found that lack of JAK1 was sufficient to block the constitutive activation of STAT3. The collective results from the various lines of investigation showed that it is, in principle, feasible to manipulate genes in a ligand-controlled manner in neoplastic mammary epithelial cells, even when cancer cells acquire a state of cellular plasticity that may no longer support the expression of the MMTV-Flp transgene.

Antineoplastic effect of iodine and iodide in dimethylbenz[a]anthracene-induced mammary tumors: association between lactoperoxidase and estrogen-adduct production

2011 ◽  
Vol 18 (4) ◽  
pp. 529-539 ◽  
Author(s):  
Ofelia Soriano ◽  
Guadalupe Delgado ◽  
Brenda Anguiano ◽  
Pavel Petrosyan ◽  
Edith D Molina-Servín ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Dna Adducts ◽  
Mammary Cancer ◽  
Mammary Tumors ◽  
Molecular Iodine ◽  
Mammary Tissue ◽  
Normal Mammary Gland ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Antineoplastic Effect

Several groups, including ours, have reported that iodine exhibited antiproliferative and apoptotic effects in various cancer cells only if this element is supplemented as molecular iodine, or as iodide, to cells that are able to oxidize it with the enzyme thyroperoxidase. In this study, we analyzed the effect of various concentrations of iodine and/or iodide in the dimethylbenz[a]anthracene (DMBA) mammary cancer model in rats. The results show that 0.1% iodine or iodide increases the expression of peroxisome proliferator-activated receptor type γ (PPARγ), triggering caspase-mediated apoptosis pathways in damaged mammary tissue (DMBA-treated mammary gland) as well as in frank mammary tumors, but not in normal mammary gland. DMBA treatment induces the expression of lactoperoxidase, which participates in the antineoplastic effect of iodide and could be involved in the pro-neoplastic effect of estrogens, increasing the formation of DNA adducts. In conclusion, our results show that a supplement of 0.1% molecular iodine/potassium iodide (0.05/0.05%) exert antineoplastic effects, preventing estrogen-induced DNA adducts and inducing apoptosis through PPARγ/caspases in pre-cancer and cancerous cells. Since this iodine concentration does not modify the cytology (histology, apoptosis rate) or physiology (triiodothyronine and thyrotropin) of the thyroid gland, we propose that it be considered as an adjuvant treatment for premenopausal mammary cancer.

165. LOCATION OF ACTIVE TGFB1 IN THE MAMMARY GLAND DURING DIFFERENT STAGES OF DEVELOPMENT

2010 ◽  
Vol 22 (9) ◽  
pp. 83
Author(s):  
X. Sun ◽  
S. A. Robertson ◽  
W. V. Ingman
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Developmental Stage ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Biological Effects ◽  
In Situ Hybridisation ◽  
Late Pregnancy ◽  
Qualitative Assessment ◽  
Normal Mammary Gland

Development of the mammary gland involves complex interactions between epithelial and stromal cells under the influence of hormones and cytokines. Transforming growth factor beta 1 (TGFB1) is a multi-functional cytokine that we have reported to be essential for normal mammary gland development. TGFB1 is produced and secreted as part of a latent complex, and requires activation at the site of action to have biological effects. In situ hybridisation studies have shown mRNA encoding Tgfb1 is mainly expressed by mammary epithelium; however, the expression pattern of active TGFB1 in the mammary gland during different developmental stages is still unclear. Mammary gland tissue was collected from mice at puberty (5 weeks old), adult diestrus and late pregnancy (day 18 pc). Frozen sections were stained with antibody specifically reactive with active TGFB1 protein (not latent TGFB1 or other TGFB isoforms) for immunofluorescent analysis. Qualitative assessment of the staining revealed different patterns of active TGFB1 localisation depending on developmental stage. The strongest expression of active TGFB1 was observed in the mammary gland at diestrus compared to puberty and pregnancy. At diestrus, active TGFB1 was located around the surface of mammary epithelial cells. The staining was heterogeneous, with distinct zones of active TGFB1 accumulated around some but not all epithelial cells. During puberty, active TGFB1 was observed only within the lumen of the ducts. During late pregnancy, TGFB1 was homogenously distributed within the alveolar epithelium. The different patterns of active TGFB1 observed during puberty, diestrus and pregnancy suggest that TGFB1 has different roles in the mammary gland dependent on developmental stage.

scholarly journals Estrogen receptor signaling is reprogrammed during breast tumorigenesis

2019 ◽  
Vol 116 (23) ◽  
pp. 11437-11443 ◽  
Author(s):  
David Chi ◽  
Hari Singhal ◽  
Lewyn Li ◽  
Tengfei Xiao ◽  
Weihan Liu ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Estrogen Receptor ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Breast Tumors ◽  
Normal Mammary Gland ◽  
Breast Tumorigenesis ◽  
Effective Prevention ◽  
Significant Difference ◽  
Estrogen Stimulation

Limited knowledge of the changes in estrogen receptor (ER) signaling during the transformation of the normal mammary gland to breast cancer hinders the development of effective prevention and treatment strategies. Differences in estrogen signaling between normal human primary breast epithelial cells and primary breast tumors obtained immediately following surgical excision were explored. Transcriptional profiling of normal ER+ mature luminal mammary epithelial cells and ER+ breast tumors revealed significant difference in the response to estrogen stimulation. Consistent with these differences in gene expression, the normal and tumor ER cistromes were distinct and sufficient to segregate normal breast tissues from breast tumors. The selective enrichment of the DNA binding motif GRHL2 in the breast cancer-specific ER cistrome suggests that it may play a role in the differential function of ER in breast cancer. Depletion of GRHL2 resulted in altered ER binding and differential transcriptional responses to estrogen stimulation. Furthermore, GRHL2 was demonstrated to be essential for estrogen-stimulated proliferation of ER+ breast cancer cells. DLC1 was also identified as an estrogen-induced tumor suppressor in the normal mammary gland with decreased expression in breast cancer. In clinical cohorts, loss of DLC1 and gain of GRHL2 expression are associated with ER+ breast cancer and are independently predictive for worse survival. This study suggests that normal ER signaling is lost and tumor-specific ER signaling is gained during breast tumorigenesis. Unraveling these changes in ER signaling during breast cancer progression should aid the development of more effective prevention strategies and targeted therapeutics.

scholarly journals Malignancy Associated MicroRNA Expression Changes in Canine Mammary Cancer of Different Malignancies

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Marie-Charlotte von Deetzen ◽  
Bernd T. Schmeck ◽  
Achim D. Gruber ◽  
Robert Klopfleisch
Keyword(s):  
Mammary Gland ◽  
Lymph Node ◽  
Mirna Expression ◽  
Mammary Cancer ◽  
Normal Mammary Gland ◽  
Primary Tumors ◽  
Normal Gland ◽  
Metastatic Cells ◽  
Node Metastases ◽  
Nucleolar Rna

MicroRNA has been suspected to be generally involved in carcinogenesis since their first description. A first study supported this assumption for canine mammary tumors when miRNA expression was compared to normal gland. The present study extends these results by comparing the expression of 16 microRNA (miRNA) and 4 small nucleolar RNA (snoRNA) in tumors of different malignancy, for example, adenomas, nonmetastasizing and metastasizing carcinomas as well as lymph node metastases, with each other and with normal mammary gland. All neoplastic tissues differed in their miR-210 expression levels from normal gland. While metastatic cells differed in their expression of mir-29b, miR-101, mir-125a, miR-143, and miR-145 from primary tumors, the comparison of miRNA expression in primary tumors of different malignancy failed to reveal significant differences except for a significant downregulation of mir-125a in metastasizing carcinomas when compared to adenomas.

scholarly journals β3 Integrin–EGF receptor cross-talk activates p190RhoGAP in mouse mammary gland epithelial cells

2011 ◽  
Vol 22 (22) ◽  
pp. 4288-4301 ◽  
Author(s):  
Nikolas Balanis ◽  
Masaaki Yoshigi ◽  
Michael K. Wendt ◽  
William P. Schiemann ◽  
Cathleen R. Carlin
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Tyrosine Phosphorylation ◽  
Focal Adhesions ◽  
Mouse Mammary Gland ◽  
Normal Mammary Gland ◽  
Integrin Subunit ◽  
Mesenchymal Transition ◽  
Β3 Integrin ◽  
Mammary Gland Epithelial Cells

Active RhoA localizes to plasma membrane, where it stimulates formation of focal adhesions and stress fibers. RhoA activity is inhibited by p190RhoGAP following integrin-mediated cell attachment to allow sampling of new adhesive environments. p190RhoGAP is itself activated by Src-dependent tyrosine phosphorylation, which facilitates complex formation with p120RasGAP. This complex then translocates to the cell surface, where p190RhoGAP down-regulates RhoA. Here we demonstrate that the epidermal growth factor receptor (EGFR) cooperates with β3 integrin to regulate p190RhoGAP activity in mouse mammary gland epithelial cells. Adhesion to fibronectin stimulates tyrosine phosphorylation of the EGFR in the absence of receptor ligands. Use of a dominant inhibitory EGFR mutant demonstrates that fibronectin-activated EGFR recruits p120RasGAP to the cell periphery. Expression of an inactive β3 integrin subunit abolishes p190RhoGAP tyrosine phosphorylation, demonstrating a mechanistic link between β3 integrin–activated Src and EGFR regulation of the RhoA inhibitor. The β3 integrin/EGFR pathway also has a positive role in formation of filopodia. Together our data suggest that EGFR constitutes an important intrinsic migratory cue since fibronectin is a key component of the microenvironment in normal mammary gland development and breast cancer. Our data also suggest that EGFR expressed at high levels has a role in eliciting cell shape changes associated with epithelial-to-mesenchymal transition.

scholarly journals A new analytical pipeline for the study of the onset of mammary gland oncogenesis based on mammary organoid transplantation and organ clearing

2019 ◽  
Author(s):  
Emilie Lagoutte ◽  
Clémentine Villeneuve ◽  
Vincent Fraisier ◽  
Denis Krndija ◽  
Marie-Ange Deugnier ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Epithelial Cells ◽  
Primary Tumor ◽  
Early Stage ◽  
Mammary Epithelium ◽  
Size Reduction ◽  
Normal Mammary Gland ◽  
Tissue Clearing ◽  
Cell Dissemination

AbstractMetastasis formation is a multi-step process starting from the dissemination of transformed carcinomatous cells from the primary tumor and could occur at a very early stage of oncogenesis, before primary tumor detection. The adult mammary gland provides a unique model to investigate epithelial cell dissemination processes. Tissue clearing techniques allow imaging samples of large volume. uDSICO clearing, one of the latest tissue clearing technique developed, provides optical imaging of whole organ due to organ clearing and tissue size reduction. We wanted to take advantage of this technique to study rare events occurring in vivo.Here, we have established a new analytical pipeline exploiting the regenerative properties of the mammary epithelium following orthotropic transplantation of organoids together with the uDISCO organ size reduction and clearing method to study early cell dissemination in the mammary gland. As proof of concept, we analyzed the localization of epithelial cells overexpressing the oncogenic protein atypical protein kinase C iota (aPKCi+) in the normal mammary gland and we were able to visualize epithelial aPKCi+ cells, surrounded by normal epithelial cells, escaping from the normal mammary epithelium and disseminating into the surrounding stroma.

scholarly journals MicroRNA in the ovine mammary gland during early pregnancy: spatial and temporal expression of miR-21, miR-205, and miR-200

2013 ◽  
Vol 45 (4) ◽  
pp. 151-161 ◽  
Author(s):  
Laurent Galio ◽  
Stéphanie Droineau ◽  
Patrick Yeboah ◽  
Hania Boudiaf ◽  
Stephan Bouet ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Mammary Gland ◽  
Epithelial Cells ◽  
Early Pregnancy ◽  
Mammary Epithelial Cells ◽  
Mammary Epithelial ◽  
Normal Mammary Gland ◽  
Basal Cells ◽  
Pregnancy And Lactation ◽  
Luminal Cells

The mammary gland undergoes extensive remodeling between the beginning of pregnancy and lactation; this involves cellular processes including cell proliferation, differentiation, and apoptosis, all of which are under the control of numerous regulators. To unravel the role played by miRNA, we describe here 47 new ovine miRNA cloned from mammary gland in early pregnancy displaying strong similarities with those already identified in the cow, human, or mouse. A microarray study of miRNA variations in the adult ovine mammary gland during pregnancy and lactation showed that 100 miRNA are regulated according to three principal patterns of expression: a decrease in early pregnancy, a peak at midpregnancy, or an increase throughout late pregnancy and lactation. One miRNA displaying each pattern (miR-21, miR-205, and miR-200b) was analyzed by qRT-PCR. Variations in expression were confirmed for all three miRNA. Using in situ hybridization, we detected both miR-21 and miR-200 in luminal mammary epithelial cells when expressed, whereas miR-205 was expressed in basal cells during the first half of pregnancy and then in luminal cells during the second half. We therefore conclude that miR-21 is strongly expressed in the luminal cells of the normal mammary gland during early pregnancy when extensive cell proliferation occurs. In addition, we show that miR-205 and miR-200 are coexpressed in luminal cells, but only during the second half of pregnancy. These two miRNA may cooperate to maintain epithelial status by repressing an EMT-like program, to achieve and preserve the secretory phenotype of mammary epithelial cells.

scholarly journals T2* relaxation times of intraductal murine mammary cancer, invasive mammary cancer, and normal mammary gland

2012 ◽  
Vol 39 (3) ◽  
pp. 1309-1313 ◽  
Author(s):  
Elizabeth Hipp ◽  
Xiaobing Fan ◽  
Sanaz A. Jansen ◽  
Erica J. Markiewicz ◽  
James Vosicky ◽  
...  
Keyword(s):  
Mammary Gland ◽  
Mammary Cancer ◽  
Relaxation Times ◽  
Normal Mammary Gland ◽  
T2 Relaxation

scholarly journals Mammary gland serotonin regulates parathyroid hormone-related protein and other bone-related signals

2012 ◽  
Vol 302 (8) ◽  
pp. E1009-E1015 ◽  
Author(s):  
Laura L. Hernandez ◽  
Karen A. Gregerson ◽  
Nelson D. Horseman
Keyword(s):  
Breast Cancer ◽  
Mammary Gland ◽  
Parathyroid Hormone ◽  
Epithelial Cells ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Related Protein ◽  
Bone Demineralization ◽  
Parathyroid Hormone Related Protein ◽  
Pthrp Expression

Breast cells drive bone demineralization during lactation and metastatic cancers. A shared mechanism among these physiological and pathological states is endocrine secretion of parathyroid hormone-related protein (PTHrP), which acts through osteoblasts to stimulate osteoclastic bone demineralization. The regulation of PTHrP has not been accounted for fully by any conventional mammotropic stimuli or tumor growth factors. Serotonin (5-HT) synthesis within breast epithelial cells is induced during lactation and in advancing breast cancer. Here we report that serotonin deficiency (knockout of tryptophan hydroxylase-1) results in a reduction of mammary PTHrP expression during lactation, which is rescued by restoring 5-HT synthesis. 5-HT induced PTHrP expression in lactogen-primed mammary epithelial cells from either mouse or cow. In human breast cancer cells 5-HT induced both PTHrP and the metastasis-associated transcription factor Runx2/Cbfa1. Based on receptor expression and pharmacological evidence, the 5-HT2 receptor type was implicated as being critical for induction of PTHrP and Runx2. These results connect 5-HT synthesis to the induction of bone-regulating factors in the normal mammary gland and in breast cancer cells.

scholarly journals Vimentin plays a functional role in mammary gland regeneration

2017 ◽  
Author(s):  
Reetta Virtakoivu ◽  
Emilia Peuhu ◽  
Anja Mai ◽  
Anni Wärri ◽  
Johanna Ivaska
Keyword(s):  
Breast Cancer ◽  
Mammary Gland ◽  
Epithelial Cell ◽  
Epithelial Cells ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Mammary Epithelial Cells ◽  
Mouse Mammary Gland ◽  
Mammary Epithelial ◽  
Basal Epithelial Cell

AbstractIn the mammary gland, vimentin intermediate filaments are expressed in stromal cells and in basal epithelial cell populations including gland-reconstituting mammary stem cells (MaSC), with largely undefined functions. Here, we studied how vimentin deficiency affects mouse mammary gland development. Our results demonstrate that in adult vimentin knockout mice (Vim-/-) mammary ductal outgrowth is delayed. The adult Vim-/- glands are characterised by dilated ducts, an imbalance in the proportion of basal to luminal mammary epithelial cells and a reduction in cells expressing Slug (Snai2), an established MaSC regulator. All of these features are indicative of reduced progenitor cell activity. Accordingly, isolated Vim-/- mammary epithelial cells display reduced capacity to form mammospheres, and altered organoid structure, compared to wt counterparts, when plated in a 3D matrix in vitro. Importantly, altered basal epithelial cell number translates into defects in Vim-/- mammary gland regeneration in vivo in cleared fat pad transplantation studies. Furthermore, we show that vimentin contributes to stem-like cell properties in triple negative MDA-MB-231 breast cancer cells, wherein vimentin depletion reduces tumorsphere formation and alters expression of breast cancer stem cell-associated surface markers. Together, our findings identify vimentin as a positive regulator of stemness in the developing mouse mammary gland and in breast cancer cells.

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