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Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry

In addition to their well-defined roles in replenishing depleted endoplasmic reticulum (ER) Ca(2+) reserves, molecular components of the store-operated Ca(2+) entry pathway regulate breast cancer metastasis. A process implicated in cancer metastasis that describes the conversion to a more invasive p...

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Autores principales: Davis, Felicity M., Peters, Amelia A., Grice, Desma M., Cabot, Peter J., Parat, Marie-Odile, Roberts-Thomson, Sarah J., Monteith, Gregory R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3364242/
https://www.ncbi.nlm.nih.gov/pubmed/22666335
http://dx.doi.org/10.1371/journal.pone.0036923
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author Davis, Felicity M.
Peters, Amelia A.
Grice, Desma M.
Cabot, Peter J.
Parat, Marie-Odile
Roberts-Thomson, Sarah J.
Monteith, Gregory R.
author_facet Davis, Felicity M.
Peters, Amelia A.
Grice, Desma M.
Cabot, Peter J.
Parat, Marie-Odile
Roberts-Thomson, Sarah J.
Monteith, Gregory R.
author_sort Davis, Felicity M.
collection PubMed
description In addition to their well-defined roles in replenishing depleted endoplasmic reticulum (ER) Ca(2+) reserves, molecular components of the store-operated Ca(2+) entry pathway regulate breast cancer metastasis. A process implicated in cancer metastasis that describes the conversion to a more invasive phenotype is epithelial-mesenchymal transition (EMT). In this study we show that EGF-induced EMT in MDA-MB-468 breast cancer cells is associated with a reduction in agonist-stimulated and store-operated Ca(2+) influx, and that MDA-MB-468 cells prior to EMT induction have a high level of non-stimulated Ca(2+) influx. The potential roles for specific Ca(2+) channels in these pathways were assessed by siRNA-mediated silencing of ORAI1 and transient receptor potential canonical type 1 (TRPC1) channels in MDA-MB-468 breast cancer cells. Non-stimulated, agonist-stimulated and store-operated Ca(2+) influx were significantly inhibited with ORAI1 silencing. TRPC1 knockdown attenuated non-stimulated Ca(2+) influx in a manner dependent on Ca(2+) influx via ORAI1. TRPC1 silencing was also associated with reduced ERK1/2 phosphorylation and changes in the rate of Ca(2+) release from the ER associated with the inhibition of the sarco/endoplasmic reticulum Ca(2+)-ATPase (time to peak [Ca(2+)](CYT) = 188.7±34.6 s (TRPC1 siRNA) versus 124.0±9.5 s (non-targeting siRNA); P<0.05). These studies indicate that EMT in MDA-MB-468 breast cancer cells is associated with a pronounced remodeling of Ca(2+) influx, which may be due to altered ORAI1 and/or TRPC1 channel function. Our findings also suggest that TRPC1 channels in MDA-MB-468 cells contribute to ORAI1-mediated Ca(2+) influx in non-stimulated cells.
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spelling pubmed-33642422012-06-04 Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry Davis, Felicity M. Peters, Amelia A. Grice, Desma M. Cabot, Peter J. Parat, Marie-Odile Roberts-Thomson, Sarah J. Monteith, Gregory R. PLoS One Research Article In addition to their well-defined roles in replenishing depleted endoplasmic reticulum (ER) Ca(2+) reserves, molecular components of the store-operated Ca(2+) entry pathway regulate breast cancer metastasis. A process implicated in cancer metastasis that describes the conversion to a more invasive phenotype is epithelial-mesenchymal transition (EMT). In this study we show that EGF-induced EMT in MDA-MB-468 breast cancer cells is associated with a reduction in agonist-stimulated and store-operated Ca(2+) influx, and that MDA-MB-468 cells prior to EMT induction have a high level of non-stimulated Ca(2+) influx. The potential roles for specific Ca(2+) channels in these pathways were assessed by siRNA-mediated silencing of ORAI1 and transient receptor potential canonical type 1 (TRPC1) channels in MDA-MB-468 breast cancer cells. Non-stimulated, agonist-stimulated and store-operated Ca(2+) influx were significantly inhibited with ORAI1 silencing. TRPC1 knockdown attenuated non-stimulated Ca(2+) influx in a manner dependent on Ca(2+) influx via ORAI1. TRPC1 silencing was also associated with reduced ERK1/2 phosphorylation and changes in the rate of Ca(2+) release from the ER associated with the inhibition of the sarco/endoplasmic reticulum Ca(2+)-ATPase (time to peak [Ca(2+)](CYT) = 188.7±34.6 s (TRPC1 siRNA) versus 124.0±9.5 s (non-targeting siRNA); P<0.05). These studies indicate that EMT in MDA-MB-468 breast cancer cells is associated with a pronounced remodeling of Ca(2+) influx, which may be due to altered ORAI1 and/or TRPC1 channel function. Our findings also suggest that TRPC1 channels in MDA-MB-468 cells contribute to ORAI1-mediated Ca(2+) influx in non-stimulated cells. Public Library of Science 2012-05-30 /pmc/articles/PMC3364242/ /pubmed/22666335 http://dx.doi.org/10.1371/journal.pone.0036923 Text en Davis et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Davis, Felicity M.
Peters, Amelia A.
Grice, Desma M.
Cabot, Peter J.
Parat, Marie-Odile
Roberts-Thomson, Sarah J.
Monteith, Gregory R.
Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry
title Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry
title_full Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry
title_fullStr Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry
title_full_unstemmed Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry
title_short Non-Stimulated, Agonist-Stimulated and Store-Operated Ca(2+) Influx in MDA-MB-468 Breast Cancer Cells and the Effect of EGF-Induced EMT on Calcium Entry
title_sort non-stimulated, agonist-stimulated and store-operated ca(2+) influx in mda-mb-468 breast cancer cells and the effect of egf-induced emt on calcium entry
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3364242/
https://www.ncbi.nlm.nih.gov/pubmed/22666335
http://dx.doi.org/10.1371/journal.pone.0036923
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