Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure

Autores
Español, Alejandro Javier; Sanchez, Yamila; Volpi, Sofia
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: Triple-negative (TN) breast cancer, the most aggressive subtype of breast cancer, is usually treated with high doses of paclitaxel (PX), which induces resistance. To prevent this adverse effect, metronomic chemotherapy based on administering low doses of PX plus carbachol (Carb), a muscarinic acetylcholine receptor (mAChR) agonist, has emerged as an alternative. Other acetylcholine receptors also present in breast tissue are nicotinic ones. When activated by nicotine (Nic), these receptors can decrease the effectiveness of conventional chemotherapy.However, whether metronomic chemotherapy with PX and Carb is affected by Nic has not yet been described. This study aimed to determine the efficacy of metronomic chemotherapy with PX and Carb in human breast tumor MDA-MB-231 cells in the presence or absence of Nic and assess the intermediaries involved. Methods: Cell viability and proliferation were determined using colorimetric assays with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue. Nitrite levels in cell supernatant were determined using Griess reagent. The expression of proteins was determined by western blot assays. Apoptosis/necrosis and the proportion of cancer stem cells (CSC) were determined by flow cytometry. Mammosphere-forming units were determined with anchorage-free growth assays. Results: The metronomic chemotherapy combining PX and Carb effectively inhibited the viability of TN MDA-MB-231 cells in the presence and absence of Nic. These effects were mediated by the activation of mAChRs, triggering signaling pathways dependent on several kinases. These mediators induce increased expression of the inducible isoform of nitric oxide synthase (NOS). Only the inducible and endothelial isoforms were expressed in these cells, and their activity was increased by the metronomic chemotherapy with PX and Carb. Nitric oxide (NO), a product of NOS activity, may contribute to the observed increase in apoptosis. We also observed an increased sensitivity to PX in the residual cells after the metronomic chemotherapy, as well as a decrease in mammosphere-forming units and CSC proportion. We also determined a decrease in the expression of stemness proteins such as ATP-binding cassette super-family G member 2 (ABCG2), sex-determining region Y-box 2 (SOX2), octamer-binding transcription factor 4 (OCT-4), and Nanog. Conclusions: Metronomic chemotherapy combining PX with Carb was selective for MDA-MB-231 cells and increased their sensitivity to conventional chemotherapy in the presence and absence of Nic, indicating that it could be a useful neoadjuvant strategy for the treatment of TN breast tumors.
Fil: Español, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina
Fil: Sanchez, Yamila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina
Fil: Volpi, Sofia. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Farmacología; Argentina
Materia
BREAST
CANCER
METRONOMIC
NICOTINE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/291262
Acceder
id CONICETDig_0bfec57648b0c7a26410c75572764adc
oai_identifier_str oai:ri.conicet.gov.ar:11336/291262
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine ExposureEspañol, Alejandro JavierSanchez, YamilaVolpi, SofiaBREASTCANCERMETRONOMICNICOTINEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: Triple-negative (TN) breast cancer, the most aggressive subtype of breast cancer, is usually treated with high doses of paclitaxel (PX), which induces resistance. To prevent this adverse effect, metronomic chemotherapy based on administering low doses of PX plus carbachol (Carb), a muscarinic acetylcholine receptor (mAChR) agonist, has emerged as an alternative. Other acetylcholine receptors also present in breast tissue are nicotinic ones. When activated by nicotine (Nic), these receptors can decrease the effectiveness of conventional chemotherapy.However, whether metronomic chemotherapy with PX and Carb is affected by Nic has not yet been described. This study aimed to determine the efficacy of metronomic chemotherapy with PX and Carb in human breast tumor MDA-MB-231 cells in the presence or absence of Nic and assess the intermediaries involved. Methods: Cell viability and proliferation were determined using colorimetric assays with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue. Nitrite levels in cell supernatant were determined using Griess reagent. The expression of proteins was determined by western blot assays. Apoptosis/necrosis and the proportion of cancer stem cells (CSC) were determined by flow cytometry. Mammosphere-forming units were determined with anchorage-free growth assays. Results: The metronomic chemotherapy combining PX and Carb effectively inhibited the viability of TN MDA-MB-231 cells in the presence and absence of Nic. These effects were mediated by the activation of mAChRs, triggering signaling pathways dependent on several kinases. These mediators induce increased expression of the inducible isoform of nitric oxide synthase (NOS). Only the inducible and endothelial isoforms were expressed in these cells, and their activity was increased by the metronomic chemotherapy with PX and Carb. Nitric oxide (NO), a product of NOS activity, may contribute to the observed increase in apoptosis. We also observed an increased sensitivity to PX in the residual cells after the metronomic chemotherapy, as well as a decrease in mammosphere-forming units and CSC proportion. We also determined a decrease in the expression of stemness proteins such as ATP-binding cassette super-family G member 2 (ABCG2), sex-determining region Y-box 2 (SOX2), octamer-binding transcription factor 4 (OCT-4), and Nanog. Conclusions: Metronomic chemotherapy combining PX with Carb was selective for MDA-MB-231 cells and increased their sensitivity to conventional chemotherapy in the presence and absence of Nic, indicating that it could be a useful neoadjuvant strategy for the treatment of TN breast tumors.Fil: Español, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Sanchez, Yamila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; ArgentinaFil: Volpi, Sofia. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Farmacología; ArgentinaTech Science Press2025-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/291262Español, Alejandro Javier; Sanchez, Yamila; Volpi, Sofia; Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure; Tech Science Press; Biocell; 49; 8; 8-2025; 1449-14801667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/biocell/v49n8/63619info:eu-repo/semantics/altIdentifier/doi/10.32604/biocell.2025.068034info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-08-25T14:53:57Zoai:ri.conicet.gov.ar:11336/291262instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-08-25 14:53:58.044CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
title Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
spellingShingle Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
Español, Alejandro Javier
BREAST
CANCER
METRONOMIC
NICOTINE
title_short Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
title_full Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
title_fullStr Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
title_full_unstemmed Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
title_sort Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure
dc.creator.none.fl_str_mv Español, Alejandro Javier
Sanchez, Yamila
Volpi, Sofia
author Español, Alejandro Javier
author_facet Español, Alejandro Javier
Sanchez, Yamila
Volpi, Sofia
author_role author
author2 Sanchez, Yamila
Volpi, Sofia
author2_role author
author
dc.subject.none.fl_str_mv BREAST
CANCER
METRONOMIC
NICOTINE
topic BREAST
CANCER
METRONOMIC
NICOTINE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Background: Triple-negative (TN) breast cancer, the most aggressive subtype of breast cancer, is usually treated with high doses of paclitaxel (PX), which induces resistance. To prevent this adverse effect, metronomic chemotherapy based on administering low doses of PX plus carbachol (Carb), a muscarinic acetylcholine receptor (mAChR) agonist, has emerged as an alternative. Other acetylcholine receptors also present in breast tissue are nicotinic ones. When activated by nicotine (Nic), these receptors can decrease the effectiveness of conventional chemotherapy.However, whether metronomic chemotherapy with PX and Carb is affected by Nic has not yet been described. This study aimed to determine the efficacy of metronomic chemotherapy with PX and Carb in human breast tumor MDA-MB-231 cells in the presence or absence of Nic and assess the intermediaries involved. Methods: Cell viability and proliferation were determined using colorimetric assays with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue. Nitrite levels in cell supernatant were determined using Griess reagent. The expression of proteins was determined by western blot assays. Apoptosis/necrosis and the proportion of cancer stem cells (CSC) were determined by flow cytometry. Mammosphere-forming units were determined with anchorage-free growth assays. Results: The metronomic chemotherapy combining PX and Carb effectively inhibited the viability of TN MDA-MB-231 cells in the presence and absence of Nic. These effects were mediated by the activation of mAChRs, triggering signaling pathways dependent on several kinases. These mediators induce increased expression of the inducible isoform of nitric oxide synthase (NOS). Only the inducible and endothelial isoforms were expressed in these cells, and their activity was increased by the metronomic chemotherapy with PX and Carb. Nitric oxide (NO), a product of NOS activity, may contribute to the observed increase in apoptosis. We also observed an increased sensitivity to PX in the residual cells after the metronomic chemotherapy, as well as a decrease in mammosphere-forming units and CSC proportion. We also determined a decrease in the expression of stemness proteins such as ATP-binding cassette super-family G member 2 (ABCG2), sex-determining region Y-box 2 (SOX2), octamer-binding transcription factor 4 (OCT-4), and Nanog. Conclusions: Metronomic chemotherapy combining PX with Carb was selective for MDA-MB-231 cells and increased their sensitivity to conventional chemotherapy in the presence and absence of Nic, indicating that it could be a useful neoadjuvant strategy for the treatment of TN breast tumors.
Fil: Español, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina
Fil: Sanchez, Yamila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Centro de Estudios Farmacológicos y Botánicos. Universidad de Buenos Aires. Facultad de Medicina. Centro de Estudios Farmacológicos y Botánicos; Argentina
Fil: Volpi, Sofia. Universidad de Buenos Aires. Facultad de Medicina. Departamento de Farmacología; Argentina
description Background: Triple-negative (TN) breast cancer, the most aggressive subtype of breast cancer, is usually treated with high doses of paclitaxel (PX), which induces resistance. To prevent this adverse effect, metronomic chemotherapy based on administering low doses of PX plus carbachol (Carb), a muscarinic acetylcholine receptor (mAChR) agonist, has emerged as an alternative. Other acetylcholine receptors also present in breast tissue are nicotinic ones. When activated by nicotine (Nic), these receptors can decrease the effectiveness of conventional chemotherapy.However, whether metronomic chemotherapy with PX and Carb is affected by Nic has not yet been described. This study aimed to determine the efficacy of metronomic chemotherapy with PX and Carb in human breast tumor MDA-MB-231 cells in the presence or absence of Nic and assess the intermediaries involved. Methods: Cell viability and proliferation were determined using colorimetric assays with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue. Nitrite levels in cell supernatant were determined using Griess reagent. The expression of proteins was determined by western blot assays. Apoptosis/necrosis and the proportion of cancer stem cells (CSC) were determined by flow cytometry. Mammosphere-forming units were determined with anchorage-free growth assays. Results: The metronomic chemotherapy combining PX and Carb effectively inhibited the viability of TN MDA-MB-231 cells in the presence and absence of Nic. These effects were mediated by the activation of mAChRs, triggering signaling pathways dependent on several kinases. These mediators induce increased expression of the inducible isoform of nitric oxide synthase (NOS). Only the inducible and endothelial isoforms were expressed in these cells, and their activity was increased by the metronomic chemotherapy with PX and Carb. Nitric oxide (NO), a product of NOS activity, may contribute to the observed increase in apoptosis. We also observed an increased sensitivity to PX in the residual cells after the metronomic chemotherapy, as well as a decrease in mammosphere-forming units and CSC proportion. We also determined a decrease in the expression of stemness proteins such as ATP-binding cassette super-family G member 2 (ABCG2), sex-determining region Y-box 2 (SOX2), octamer-binding transcription factor 4 (OCT-4), and Nanog. Conclusions: Metronomic chemotherapy combining PX with Carb was selective for MDA-MB-231 cells and increased their sensitivity to conventional chemotherapy in the presence and absence of Nic, indicating that it could be a useful neoadjuvant strategy for the treatment of TN breast tumors.
publishDate 2025
dc.date.none.fl_str_mv 2025-08
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/291262
Español, Alejandro Javier; Sanchez, Yamila; Volpi, Sofia; Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure; Tech Science Press; Biocell; 49; 8; 8-2025; 1449-1480
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/291262
identifier_str_mv Español, Alejandro Javier; Sanchez, Yamila; Volpi, Sofia; Metronomic Chemotherapy Response in MDA-MB-231 Triple-Negative Breast Cancer Cells under Nicotine Exposure; Tech Science Press; Biocell; 49; 8; 8-2025; 1449-1480
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/biocell/v49n8/63619
info:eu-repo/semantics/altIdentifier/doi/10.32604/biocell.2025.068034
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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