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Immunoglobulin G from Breast Cancer Patients in Stage I Stimulates Muscarinic Acetylcholine Receptors in MCF7 Cells and Induces Proliferation. Participation of Nitric Oxide Synthase-Derived Nitric Oxide

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Abstract

Introduction

Muscarinic acetylcholine receptors (mAChR) belong to the G-protein-coupled receptor family and are extensively expressed in most cells in mammals. We had reported the expression of mAChR in murine and human breast tumors.

Methods

The presence of antibodies in the sera of patients with different tumors directed against self-proteins has been recently described. In this work, we investigated the presence of autoantibodies against mAChR in the sera of breast cancer patients in stage I (T1N0Mx-IgG). IgG purification was performed by affinity chromatography in protein G-agarose. We also studied the ability of these antibodies to modulate the proliferation of MCF-7 breast tumor cells by the MTS colorimetric assay. The ability of T1N0Mx-IgG to stimulate muscarinic signaling pathway via nitric oxide synthase was tested by Griess reaction.

Results

We demonstrated M3 and M4 receptors expression in MCF-7 cells. T1N0Mx-IgG promotes cell proliferation, mimicking the action of the muscarinic agonist carbachol. This effect was preferentially due to M3 receptor activation in tumor cells via phospholipase C-induced nitric oxide liberation by calcium-dependent nitric oxide synthases. IgG from control patients was unable to produce this effect.

Discussion

IgG from patients with breast cancer in early stages could be promoting tumor progression by muscarinic activation, and its presence could be determining the prognosis of this illness.

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Acknowledgments

The authors want to thank Mrs. María Ester Castro for her excellent technical assistance. This work was supported by grants from University of Buenos Aires (UBACYT MO064) and from the National Agency for Scientific and Technology Promotion (ANPCyT PICT 2006-485). Drs. Alejandro J. Español and María Elena Sales are established investigators in the National Research Council of Argentina (CONICET). Dr. Eulalia de la Torre is a fellow from CONICET.

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Authors and Affiliations

  1. Centro de Estudios Farmacológicos y Botánicos (CEFYBO)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 piso 16, CP 1121, Ciudad Autónoma de Buenos Aires, Argentina

    María Pía Negroni, Alejandro J. Español, Laura T. Pelegrina, Eulalia de la Torre & María Elena Sales

  2. Instituto de Oncología Angel H. Roffo, Universidad de Buenos Aires, Ciudad Autónoma de Buenos Aires, Argentina

    Gabriel L. Fiszman, María E. Azar & Carlos Cresta Morgado

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  1. María Pía Negroni
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  2. Gabriel L. Fiszman
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  5. Alejandro J. Español
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  7. Eulalia de la Torre
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  8. María Elena Sales
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Correspondence to María Elena Sales.

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Negroni, M.P., Fiszman, G.L., Azar, M.E. et al. Immunoglobulin G from Breast Cancer Patients in Stage I Stimulates Muscarinic Acetylcholine Receptors in MCF7 Cells and Induces Proliferation. Participation of Nitric Oxide Synthase-Derived Nitric Oxide. J Clin Immunol 30, 474–484 (2010). https://doi.org/10.1007/s10875-010-9370-0

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