scholarly journals Modulation of Epithelial Mesenchymal Transition after AGTR-1 Gene Edition by Crispr/Cas9 and Losartan Treatment in Mammary Tumor Cell Line: A Comparative Study between Human and Canine Species

Life ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1427
Author(s):  
Marina Gobbe Moschetta-Pinheiro ◽  
Jucimara Colombo ◽  
Bianca Lara Venâncio de Godoy ◽  
Julia Ferreira Balan ◽  
Bianca Carlos Nascimento ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Mammary Tumor ◽  
Tumor Cell Line ◽  
Type I ◽  
Tumor Type ◽  
Mammary Tumor Cell ◽  
Mammary Tumor Cell Line ◽  
Mesenchymal Transition ◽  
Parp 1

Breast cancer is the most prevalent tumor type among women and female dogs. Tumor malignancy is characterized by the epithelial-to-mesenchymal transition (EMT) which leads to the metastasis formation. The inhibition of angiotensin II type I receptor (AGTR1) by an antagonist such as losartan can suppress angiogenesis, consequently contributing to the metastasis control. The aim of this study was to analyze the capacity of losartan and AGTR-1 gene edition to modulate the EMT process in triple negative/metastatic mammary tumor cells, compared to existing treatment protocols such as carboplatin. The cell lines CF41.Mg and MDA-MB-468, were cultured and treated with carboplatin, losartan, or submitted to AGTR-1 gene edition by CRISPR/Cas9. EMT markers and PARP-1 protein and gene expression were evaluated by immunofluorescence or immunocytochemistry and qRT-PCR, respectively. Cell migration capacity was also evaluated. For CF41.Mg and MDA-MB-468 cell lines, there was an increase in E-cadherin and a decrease in N-cadherin and PARP-1 protein and gene expression after treatment with carboplatin, losartan, both in combination and after AGTR-1 gene edition. There was a decrease in VEGF and PARP-1 protein and gene expression after AGTR-1 gene edition. Moreover, in both lines, reduction in invasion rate was observed after all treatments. Our data suggest that losartan and the gene edition of AGTR-1 by CRISPR/Cas9 were able to block the DNA repair and control the EMT process, such as carboplatin. The results in the canine species are unprecedented, as there are no data in the literature that demonstrate the action of losartan in this tumor type.

Hormonal induction of transfected genes depends on DNA topology

1990 ◽  
Vol 10 (2) ◽  
pp. 625-633
Author(s):  
B Piña ◽  
R J Haché ◽  
J Arnemann ◽  
G Chalepakis ◽  
E P Slater ◽  
...  
Keyword(s):  
Cell Line ◽  
Mammary Tumor ◽  
Binding Sites ◽  
Glucocorticoid Receptors ◽  
Hormone Receptors ◽  
Regulatory Element ◽  
Tumor Cell Line ◽  
Dna Topology ◽  
Mammary Tumor Cell ◽  
Mammary Tumor Cell Line

Plasmids containing the hormone regulatory element of mouse mammary tumor virus linked to the thymidine kinase promoter of herpes simplex virus and the reporter gene chloramphenicol acetyltransferase of Escherichia coli respond to glucocorticoids and progestins when transfected into appropriate cells. In the human mammary tumor cell line T47D, the response to progestins, but not to glucocorticoids, is highly dependent on the topology of the transfected DNA. Although negatively supercoiled plasmids respond optimally to the synthetic progestin R5020, their linearized counterparts exhibit markedly reduced progestin inducibility. This is not due to changes in the efficiency of DNA transfection, since the amount of DNA incorporated into the cell nucleus is not significantly dependent on the initial topology of the plasmids. In contrast, cotransfection experiments with glucocorticoid receptor cDNA in the same cell line show no significant influence of DNA topology on induction by dexamethasone. A similar result was obtained with fibroblasts that contain endogenous glucocorticoid receptors. When the distance between receptor-binding sites or between the binding sites and the promoter was increased, the dependence of progestin induction on DNA topology was more pronounced. In contrast to the original plasmid, these constructs also revealed a similar topological dependence for induction by glucocorticoids. The differential influence of DNA topology is not due to differences in the affinity of the two hormone receptors for DNA of various topologies, but probably reflects an influence of DNA topology on the interaction between different DNA-bound receptor molecules and between receptors and other transcription factors.

Formation of hemidesmosomes in cells of a transformed murine mammary tumor cell line and mechanisms involved in adherence of these cells to laminin and kalinin

1993 ◽  
Vol 106 (4) ◽  
pp. 1083-1102 ◽  
Author(s):  
A. Sonnenberg ◽  
A.A. de Melker ◽  
A.M. Martinez de Velasco ◽  
H. Janssen ◽  
J. Calafat ◽  
...  
Keyword(s):  
Cell Line ◽  
Mammary Tumor ◽  
Cytochalasin B ◽  
Tumor Cell Line ◽  
Metabolic Energy ◽  
Mammary Tumor Cell ◽  
Mammary Tumor Cell Line ◽  
Murine Mammary Tumor ◽  
Integrin Alpha 6 ◽  
In Cells

Keratinocytes attach to an underlying basement membrane by adhesion junctions called hemidesmosomes. We have characterized a cell line, RAC-11P/SD, established from a murine mammary tumor, which differentiates into squamous epithelium and forms well defined hemidesmosomes. These hemidesmosomes contain the integrin alpha 6 beta 4 as well as the hemidesmosomal plaque proteins BP230 and HD1 and are associated with a matrix containing kalinin and laminin. We examined how these cells adhere to laminin and to kalinin present in matrices as well as immunopurified kalinin. We show that adhesion to laminin is energy dependent but does not require an intact actin-containing cytoskeleton. The affinity for kalinin proved to be greater and binding to kalinin was still observed when cells had been treated with deoxyglucose and azide to inhibit metabolic energy. Binding to laminin (or fragment E8), but not to kalinin was partially blocked by a monoclonal antibody specific for the integrin alpha 6 subunit, and only in the initial phase of adhesion. The antibody efficiently blocked adhesion to laminin of cells treated with the microfilament disrupting drug cytochalasin B, but only partially blocked the adhesion of cytochalasin B-treated cells to kalinin, while adherence of cells treated with deoxyglucose and azide to kalinin was blocked completely. The integrin alpha 6 beta 4 is redistributed to the basal surface during adhesion and then is organized into ring-like structures when cells are bound to laminin and localized into hemidesmosomes in cells adhered to kalinin. We suggest that anti-alpha 6 hinders the binding of the alpha 6 beta 4 integrins to its ligands laminin and kalinin, but cannot prevent adhesion when clustering of the integrin has become complete. In addition, there is evidence that adhesion to kalinin is mediated by a second receptor, which associates with the actin-containing cytoskeleton. The presence of such a second receptor is suggested because the cells can spread on kalinin, but not when they have been treated with cytochalasin B. On laminin spreading does not occur, irrespective of whether cells have been treated with cytochalasin B or not. The integrin alpha 3 beta 1, which has been identified as a receptor for kalinin but not for laminin, is strongly expressed in RAC-11P/SD cells and it seems likely that this integrin is responsible for spreading of cells on kalinin.

Establishment and characterization of a new murine mammary tumor cell line, balb/c-MC

1987 ◽  
Vol 23 (11) ◽  
pp. 755-758 ◽  
Author(s):  
Junji Morimoto ◽  
Shunsuke Imai ◽  
Yuzo Taniguchi ◽  
Yoshihiko Tsubura ◽  
Howard L. Hosick
Keyword(s):  
Tumor Cell ◽  
Cell Line ◽  
Mammary Tumor ◽  
Tumor Cell Line ◽  
Mammary Tumor Cell ◽  
Mammary Tumor Cell Line ◽  
Murine Mammary Tumor

scholarly journals Rosiglitazone inhibits metastasis development of a murine mammary tumor cell line LMM3

BMC Cancer ◽  
2008 ◽  
Vol 8 (1) ◽  
Author(s):  
Gabriela Magenta ◽  
Ximena Borenstein ◽  
Romina Rolando ◽  
María Adela Jasnis
Keyword(s):  
Tumor Cell ◽  
Cell Line ◽  
Mammary Tumor ◽  
Tumor Cell Line ◽  
Mammary Tumor Cell ◽  
Mammary Tumor Cell Line ◽  
Murine Mammary Tumor ◽  
Metastasis Development
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