Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells

Supplementary figures: 1-3

Detalles Bibliográficos
Autores: Jardín, Isaac, Alvarado, Sandra, Jiménez-Velarde, Vanesa, Nieto-Felipe, Joel, López, José J., Salido, Ginés M., Smani, Tarik, Rosado, Juan A.
Tipo de recurso: conjunto de datos
Fecha de publicación:2023
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/352141
Acceso en línea:http://hdl.handle.net/10261/352141
Access Level:acceso abierto
Palabra clave:Biochemistry
Breast cancer
Cell biology
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dc.title.none.fl_str_mv Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
title Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
spellingShingle Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
Jardín, Isaac
Biochemistry
Breast cancer
Cell biology
title_short Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
title_full Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
title_fullStr Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
title_full_unstemmed Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
title_sort Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells
dc.creator.none.fl_str_mv Jardín, Isaac
Alvarado, Sandra
Jiménez-Velarde, Vanesa
Nieto-Felipe, Joel
López, José J.
Salido, Ginés M.
Smani, Tarik
Rosado, Juan A.
author Jardín, Isaac
author_facet Jardín, Isaac
Alvarado, Sandra
Jiménez-Velarde, Vanesa
Nieto-Felipe, Joel
López, José J.
Salido, Ginés M.
Smani, Tarik
Rosado, Juan A.
author_role author
author2 Alvarado, Sandra
Jiménez-Velarde, Vanesa
Nieto-Felipe, Joel
López, José J.
Salido, Ginés M.
Smani, Tarik
Rosado, Juan A.
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Ministerio de Ciencia e Innovación (España)
European Commission
Agencia Estatal de Investigación (España)
Junta de Extremadura
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Biochemistry
Breast cancer
Cell biology
topic Biochemistry
Breast cancer
Cell biology
description Supplementary figures: 1-3
publishDate 2023
dc.date.none.fl_str_mv 2023
2024
2024
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url http://hdl.handle.net/10261/352141
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Jardín, Isaac; Alvarado, Sandra; Jiménez-Velarde, Vanesa; Nieto-Felipe, Joel; López, José J.; Salido, Ginés M.; Smani, Tarik; Rosado, Juan A. Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells. https://doi.org/10.1038/s41598-023-46946-8. http://hdl.handle.net/10261/352140
https://doi.org/10.1038/s41598-023-46946-8
Sí
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dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
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spelling Supplementary figures: Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cellsJardín, IsaacAlvarado, SandraJiménez-Velarde, VanesaNieto-Felipe, JoelLópez, José J.Salido, Ginés M.Smani, TarikRosado, Juan A.BiochemistryBreast cancerCell biologySupplementary figures: 1-3Orai1 is the pore-forming subunit of the Ca2+-release activated Ca2+ channels that mediate store-operated Ca2+ entry (SOCE) in excitable and non-excitable cells. Two Orai1 forms have been identified in mammalian cells, the full-length variant Orai1α, and the short form Orai1β, lacking the N-terminal 63 amino acids. Stem cells were isolated from non-tumoral breast epithelial cells of the MCF10A cell line, and the most representative ER+ , HER2 or triple negative breast cancer cell lines MCF7, SKBR3 and MDA-MB-231, respectively. Orai and TRPC family members expression was detected by RT-PCR and Western blotting. Changes in cytosolic Ca2+ concentration were analyzed by confocal microscopy using Fluo 4 and the spheroid-forming ability and self-renewal was estimated in culture plates coated with pHEMA using a cell imaging system. Here, we have characterized the expression of Orai family members and several TRPC channels at the transcript level in breast stem cells (BSC) derived from the non-tumoral breast epithelial cell line MCF10A and breast cancer stem cells (BCSC) derived from the well-known estrogen receptor positive (ER+), HER2 and triple negative cell lines MCF7, SKBR3 and MDA-MB-231, respectively. Furthermore, we have evaluated the mammosphere formation efficiency and self-renewal of the BSC and BCSC. Next, through a combination of Orai1 knockdown by iRNA and the use of MDA-MB-231 KO cells, missing the native Orai1, transfected with plasmids encoding for either Orai1α or Orai1β, we show that Orai1 is essential for mammosphere formation and self-renewal efficiency in BCSC derived from triple negative and HER2 subtypes cell cultures, while this channel has a negligible effect in BCSC derived from ER+ cells as well as in non-tumoral BSC. Both, Orai1α, and Orai1β support SOCE in MDA-MB-231-derived BCSC with similar efficiency, as well as COX activation and mammosphere formation. These findings provide evidence of the functional role of Orai1α and Orai1β in spheroid forming efficiency and self-renewal in breast cancer stem cells.This research was supported by PID2019-104084 GB-C21, PID2019-104084 GB-C22 and PID2022-136279NB-C21 funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”, and Junta de Extremadura-Fondo Europeo de Desarrollo Regional (FEDER; Grant IB20007 and GR21008) to JAR. J.N.-F. and S.A. are supported by a contract from Ministry of Science, Innovation, and Universities, Spain and PID2019-104084 GB-C21 funded by MCIN/AEI/10.13039/501100011033, respectively.Peer reviewedNature Publishing GroupMinisterio de Ciencia, Innovación y Universidades (España)Ministerio de Ciencia e Innovación (España)European CommissionAgencia Estatal de Investigación (España)Junta de ExtremaduraConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242023info:eu-repo/semantics/datasethttp://purl.org/coar/resource_type/c_ddb1application/mswordhttp://hdl.handle.net/10261/352141reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104084GB-C21info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104084GB-C22info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-136279NB-C21info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2019-104084GB-C21Jardín, Isaac; Alvarado, Sandra; Jiménez-Velarde, Vanesa; Nieto-Felipe, Joel; López, José J.; Salido, Ginés M.; Smani, Tarik; Rosado, Juan A. Orai1α and Orai1β support calcium entry and mammosphere formation in breast cancer stem cells. https://doi.org/10.1038/s41598-023-46946-8. http://hdl.handle.net/10261/352140https://doi.org/10.1038/s41598-023-46946-8Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3521412026-05-22T06:33:51Z
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