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Heparins modulate the IFN-γ-induced production of chemokines in human breast cancer cells

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Abstract

Heparins seem to improve survival in patients with advanced malignancies independently of their anticoagulatory function. As the treatment options in advanced and metastatic breast cancer are still very limited, heparins might be an interesting addition to the existing systemic therapies. The interferon (IFN)-γ-inducible chemokines CXCL9 and CXCL10 play an essential role in the regulation of the immune milieu in malignant tumours, thereby being interesting targets for an immunological intervention. We therefore wanted to test whether heparins have an impact on the chemokines CXCL9 and CXCL10 as well as the IFN-γ signalling in human breast cancer cells in vitro. The well-established cell lines BT-474, MCF-7, SK-BR-3 and MDA-MB-231 were incubated with IFN-γ, unfractionated heparin (UFH), different low molecular weight heparins (LMWHs) and the heparin-related polyanions danaparoid and dextran sulphate. The production of CXCL9 and CXCL10 was measured by ELISA and real-time RT-PCR, the phosphorylation of signal transducer and activator of transcription (STAT) 1 was detected by an in-cell western assay and the amount of cellular bound IFN-γ was analysed by a high sensitivity ELISA. We observed that IFN-γ induced CXCL9 and CXCL10 production in MCF-7, SK-BR-3 and MDA-MB-231 cells but not in BT-474. UFH dose dependently inhibited the effect of IFN-γ on the secretion and expression of CXCL9 and CXCL10. LMWHs and heparin-related compounds differentially modulated IFN-γ-effects—the results depended on their molecular size and charge, but were independent of their anticoagulatory properties. As a reason for these heparin effects, we could show that the IFN-γ-induced phosphorylation of STAT1 was modulated by heparins, caused by an interaction with the cellular binding of IFN-γ. In conclusion, these results support the significance of the immunomodulatory properties of heparins independently of their classical anticoagulatory function. Heparin-derived sulphated polysaccharides with distinct molecular properties might thus be interesting candidates for new therapeutic strategies in breast cancer.

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Abbreviations

CD:

Cluster of differentiation

cDNA:

Complementary deoxyribonucleic acid

ELISA:

Enzyme-linked immunosorbent assay

ER:

Oestrogen-receptor

FXa:

Factor Xa

GAG:

Glycosaminoglycan

Her-2:

Human epidermal growth factor receptor-2

HS:

Heparan sulphate

IFN-γ:

Interferon-γ

Jak:

Janus kinase

LMWH:

Low molecular weight heparin

NK:

Natural killer

PR:

Progesterone-receptor

RNA:

Ribonucleic acid

RT-PCR:

Reverse transcriptase polymerase chain reaction

STAT1:

Signal transducer and activator of transcription 1

Th1:

T helper cells type 1

UFH:

Unfractionated heparin

VTE:

Venous thromboembolism

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Acknowledgments

The authors thank Stephanie Heidrich, Department of Obstetrics and Gynecology, University of Greifswald, Germany, for excellent technical assistance.

Conflict of interest

The authors have no conflict of interest.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, University of Greifswald, Sauerbruchstr., 17475, Greifswald, Germany

    Herbert Fluhr, Tina Seitz & Marek Zygmunt

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  1. Herbert Fluhr
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  2. Tina Seitz
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  3. Marek Zygmunt
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Corresponding author

Correspondence to Herbert Fluhr.

Additional information

Herbert Fluhr and Tina Seitz contributed equally to this work.

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Fluhr, H., Seitz, T. & Zygmunt, M. Heparins modulate the IFN-γ-induced production of chemokines in human breast cancer cells. Breast Cancer Res Treat 137, 109–118 (2013). https://doi.org/10.1007/s10549-012-2334-8

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  • DOI: https://doi.org/10.1007/s10549-012-2334-8

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