The metronomic combination of paclitaxel with cholinergic agonists inhibits triple negative breast tumor progression. Participation of M2 receptor subtype

Alejandro J Español; Agustina Salem; María Di Bari; Ilaria Cristofaro; Yamila Sanchez; Ada M Tata; María E Sales

doi:10.1371/journal.pone.0226450

The metronomic combination of paclitaxel with cholinergic agonists inhibits triple negative breast tumor progression. Participation of M2 receptor subtype

PLoS One. 2020 Sep 10;15(9):e0226450. doi: 10.1371/journal.pone.0226450. eCollection 2020.

Authors

Alejandro J Español^{1

2

3}, Agustina Salem^{1

2}, María Di Bari³, Ilaria Cristofaro³, Yamila Sanchez^{1

2}, Ada M Tata^{3

4}, María E Sales^{1

2}

Affiliations

¹ Center of Pharmacological and Botanical Studies (CEFYBO), CONICET, Buenos Aires, Argentine.
² Department of Pharmacology, School of Medicine, University of Buenos Aires, Buenos Aires, Argentine.
³ Department of Biology and Biotechnologies Charles Darwin, Sapienza University of Rome, Rome, Italy.
⁴ Center of Neurobiology Daniel Bovet, Sapienza University of Rome, Rome, Italy.

Abstract

Triple negative tumors are more aggressive than other breast cancer subtypes and there is a lack of specific therapeutic targets on them. Since muscarinic receptors have been linked to tumor progression, we investigated the effect of metronomic therapy employing a traditional anti-cancer drug, paclitaxel plus muscarinic agonists at low doses on this type of tumor. We observed that MDA-MB231 tumor cells express muscarinic receptors, while they are absent in the non-tumorigenic MCF-10A cell line, which was used as control. The addition of carbachol or arecaidine propargyl ester, a non-selective or a selective subtype 2 muscarinic receptor agonist respectively, plus paclitaxel reduces cell viability involving a down-regulation in the expression of ATP "binding cassette" G2 drug transporter and epidermal growth factor receptor. We also detected an inhibition of tumor cell migration and anti-angiogenic effects produced by those drug combinations in vitro and in vivo (in NUDE mice) respectively. Our findings provide substantial evidence about subtype 2 muscarinic receptors as therapeutic targets for the treatment of triple negative tumors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATP Binding Cassette Transporter, Subfamily G, Member 2 / metabolism
Administration, Metronomic
Animals
Antineoplastic Combined Chemotherapy Protocols / administration & dosage*
Arecoline / administration & dosage
Arecoline / analogs & derivatives
Carbachol / administration & dosage
Cell Line, Tumor
Cell Movement / drug effects
Cell Proliferation / drug effects
Cell Survival / drug effects
Cholinergic Agonists / administration & dosage*
Down-Regulation / drug effects
ErbB Receptors / metabolism
Female
Gene Expression Regulation, Neoplastic / drug effects
Humans
Mice
Neoplasm Proteins / metabolism
Neovascularization, Pathologic / drug therapy
Neovascularization, Pathologic / pathology
Paclitaxel / administration & dosage*
RNA, Small Interfering / metabolism
Receptor, Muscarinic M2 / agonists
Receptor, Muscarinic M2 / genetics
Receptor, Muscarinic M2 / metabolism*
Triple Negative Breast Neoplasms / blood supply
Triple Negative Breast Neoplasms / drug therapy*
Triple Negative Breast Neoplasms / pathology
Vascular Endothelial Growth Factor A / metabolism
Xenograft Model Antitumor Assays

Substances

ABCG2 protein, human
ATP Binding Cassette Transporter, Subfamily G, Member 2
CHRM2 protein, human
Cholinergic Agonists
Neoplasm Proteins
RNA, Small Interfering
Receptor, Muscarinic M2
VEGFA protein, human
Vascular Endothelial Growth Factor A
arecaidine esters
Arecoline
Carbachol
EGFR protein, human
ErbB Receptors
Paclitaxel

Grants and funding

AJE; ANPCyT PICT 2015-2017, 2396. Agencia Nacional de Promoción Científica y Tecnológica. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. MES, CONICET PIP 2015-2017, 201501 00239; Consejo Nacional de Investigaciones Cientificas y Técnicas; UBA UBACYT 2014-2017, 20020130100168BA Universidad de Buenos Aires. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.