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Hypoxia-induced nitric oxide release by luminal cells stimulates proliferation and uPA secretion of myoepithelial cells in a bicellular murine mammary tumor

  • Original Article – Cancer Research
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Abstract

Introduction

LM38 murine mammary adenocarcinoma model is formed by LM38-LP (myoepithelial and luminal), LM38-HP (luminal) and LM38-D2 (myoepithelial) cell lines. In a previous work, we had shown that LM38-HP and LM38-D2 cell lines are less malignant than the bicellular LM38-LP cell line.

Purpose

To study the role of nitric oxide (NO) as one of the mediators of functional interactions between malignant luminal and myoepithelial cells.

Methods and results

Using immunohistochemistry, in vivo iNOS expression was only detected in the luminal cells of bicellular LM38-LP and most cells of LM38-HP tumors. In cobalt-induced pseudohypoxia, LM38-LP and LM38-HP cell lines significantly increased HIF-1α and iNOS expression (Western blotting) and therefore NO production (Griess method). This increase was inhibited by the iNOS inhibitor 1400 W. On the other side, NO was not detectable in LM38-D2 cells either in basal or in pseudohypoxia. In addition, pseudohypoxia increased urokinase-type plasminogen activator (uPA) secretion by LM38-LP and LM38-HP cells and migration in the LM38-LP cell line, without modulating these properties in LM38-D2 cells (radial caseinolysis). The NO donor DETA/NONOate (500 μM) was able to increase uPA secretion and in vitro growth of LM38-D2. In agreement, 1400 W prevented in vivo growth of the myoepithelial LM38-D2 cells.

Conclusions

Hypoxia leads to an enhanced NO production by the luminal component, through HIF-1α and iNOS, which can stimulate myoepithelial cell proliferation and uPA secretion. In these new conditions, myoepithelial cells might act as an invasive forefront generating gaps that could help luminal cells to escape from the primary tumor.

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Acknowledgments

We would like to thank MSc Lina Marino, Alicia Rivelli and Silvina Romero for help with immunohistochemistry and technical assistance. Also, we are thankful to Vanina Rodriguez and Candela Martin for the provision and care of mice. This work was supported by Grants from the Agencia Nacional para la Promoción de la Ciencia y la Tecnolgía (ANPCyT, Prestamo BID PICT2010-01296), from the Universidad de Buenos Aires (UBACYT MO15 and M20020100100243) and from Concejo Nacional de Investigaciones Científicas y Técnicas (CONICET, PIP193). No founding source had any involvement in study design; the collection, analysis or interpretation of data; in the writing of the report; or in the decision to submit the article for publication.

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Author notes

  1. Martin Alejandro Krasnapolski

    Present address: Departamento de patología, Instituto de Oncología “Ángel H Roffo”, Buenos Aires, Argentina

  2. Catalina Lodillinsky

    Present address: Institut Curie Research Center, CNRS UMR 144, Paris, France

Authors and Affiliations

  1. Área Investigación, Instituto de Oncología “Ángel H Roffo”, Av. San Martín 5481, C1417DTB, Buenos Aires, Argentina

    Martin Alejandro Krasnapolski, Catalina Lodillinsky, Elisa Bal De Kier Joffé & Ana María Eiján

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  1. Martin Alejandro Krasnapolski
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  2. Catalina Lodillinsky
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Correspondence to Ana María Eiján.

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Elisa Bal De Kier Joffé and Ana María Eiján are Members of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET).

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Krasnapolski, M.A., Lodillinsky, C., Bal De Kier Joffé, E. et al. Hypoxia-induced nitric oxide release by luminal cells stimulates proliferation and uPA secretion of myoepithelial cells in a bicellular murine mammary tumor. J Cancer Res Clin Oncol 141, 1727–1738 (2015). https://doi.org/10.1007/s00432-015-1934-1

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  • DOI: https://doi.org/10.1007/s00432-015-1934-1

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