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Influence of Terminal Differentiation and PACAP on the Cytokine, Chemokine, and Growth Factor Secretion of Mammary Epithelial Cells

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Abstract

Pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide with trophic and cytoprotective effects, has been shown to affect cell survival, proliferation, and also differentiation of various cell types. The high PACAP level in the milk and its changes during lactation suggest a possible effect of PACAP on the differentiation of mammary epithelial cells. Mammary cell differentiation is regulated by hormones, growth factors, cytokines/chemokines, and angiogenic proteins. In this study, differentiation was hormonally induced by lactogenic hormones in confluent cultures of HC11 mouse mammary epithelial cells. We investigated the effect of PACAP on mammary cell differentiation as well as release of cytokines, chemokines, and growth factors. Differentiation was assessed by expression analysis of the milk protein β-casein. Differentiation significantly decreased the secretion of interferon gammainduced protein (IP)-10, “regulated upon activation normal T cell expressed and presumably secreted” (RANTES), insulin-like growth factor-binding protein (IGFBP)-3 and the epidermal growth factor receptor (EGFR) ligands, such as epidermal growth factor (EGF) and amphiregulin (AREG). The changes in the levels of IP-10 and RANTES may be relevant for the alterations in homing of T cells and B cells at different stages of mammary gland development, while the changes of the EGFR ligands may facilitate the switch from proliferative to lactating stage. PACAP did not modulate the expression of β-casein or the activity of hormone-induced pathways as determined by the analysis of phosphorylation of Akt, STAT5, and p38 MAPK. However, PACAP decreased the release of EGF and AREG from non-differentiated cells. This may influence the extracellular signal-related transactivation of EGFR in the non-differentiated mammary epithelium and is considered to have an impact on the modulation of oncogenic EGFR signaling in breast cancer.

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Abbreviations

ADAM17:

A disintegrin and metalloprotease 17

AREG:

Amphiregulin

cAMP:

Cyclic adenosine monophosphate

CTGF:

Connective tissue growth factor

EGF:

Epidermal growth factor

EGFR:

Epidermal growth factor receptor

FGF:

Fibroblast growth factor

G-CSF:

Granulocyte colony-stimulating factor

HGF:

Hepatocyte growth factor

IGF:

Insulin-like growth factor

IGFBP:

Insulin-like growth factor-binding protein

IL:

Interleukin

IL-1ra:

Interleukin 1 receptor antagonist

IP-10:

Interferon gamma-induced protein 10

JAK:

Janus kinase

M-CSF:

Macrophage colony-stimulating factor

p38 MAPK:

p38 Mitogen-activated protein kinases

PACAP:

Pituitary adenylate cyclase-activating polypeptide

PDGF:

Platelet-derived growth factor

PKA:

Protein Kinase A

PRL:

Prolactin

PTHLH:

Parathyroid hormone-like hormone

RANK-L:

Receptor activator of NF-kB ligand

RANTES:

Regulated upon activation normal T cell expressed and presumably secreted

STAT:

Signal transducer and activator of transcription

TIMP:

Tissue inhibitor of metalloproteinase

TGF:

Transforming growth factor

TNF:

Tumor necrosis factor

VEGF:

Vascular endothelial growth factor

VIP:

Vasoactive intestinal peptide

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Acknowledgments

This work was supported by PTE-MTA Lendület Program, Arimura Foundation, OTKA K104984, 4.2.2.A-11/1/KONV-2012-0024, and TAMOP 4.2.4.A/2-11-1-2012-0001 (National Excellence Program).

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Authors and Affiliations

  1. Department of Anatomy, PTE-MTA “Lendulet” PACAP Research Team, University of Pecs, Szigeti ut 12, Pecs, 7624, Hungary

    Katalin Csanaky, Andrea Tamas & Dora Reglodi

  2. Division of Medical Biochemistry, Biocenter, Innsbruck Medical University, Innrain 80, Innsbruck, 6020, Austria

    Wolfgang Doppler

  3. Department of Biochemistry and Medical Chemistry, University of Pecs, Szigeti ut 12, Pecs, 7624, Hungary

    Krisztina Kovacs

  4. Department of Medical Chemistry, University of Szeged, Dom ter 8, Szeged, 6720, Hungary

    Gabor Toth

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  1. Katalin Csanaky
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  2. Wolfgang Doppler
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  3. Andrea Tamas
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  4. Krisztina Kovacs
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  5. Gabor Toth
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  6. Dora Reglodi
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Correspondence to Dora Reglodi.

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Csanaky, K., Doppler, W., Tamas, A. et al. Influence of Terminal Differentiation and PACAP on the Cytokine, Chemokine, and Growth Factor Secretion of Mammary Epithelial Cells. J Mol Neurosci 52, 28–36 (2014). https://doi.org/10.1007/s12031-013-0193-3

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