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EGF activates PI3K-Akt and NF-κB via distinct pathways in salivary epithelial cells in Sjögren’s syndrome

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Abstract

Epidermal growth factor (EGF) exerts tropic effects on salivary epithelial cells. We examined EGF-mediated signaling pathways in the salivary epithelial cells of patients with Sjögren’s syndrome (SS). We compared the immunohistochemical expression of EGF receptor (EGF-R), phosphatidylinositol 3-kinase (PI3K), Akt and nuclear factor kappa B (NF-κB) in the labial salivary glands of SS patients (n = 6) with those of control subjects (n = 2). EGF-mediated signaling pathways were further studied in vitro (n = 3) using primary salivary epithelial cells; NF-κB p65 nuclear translocation and Akt phosphorylation were examined by immunofluorescence and western blotting, respectively. The phosphorylation of EGF-R and Akt, and the nuclear expression of NF-κB p65, were increased in situ in the salivary epithelial cells of SS patients compared with those of control subjects. Epidermal growth factor induced rapid EGF-R phosphorylation and NF-κB p65 nuclear translocation in primary salivary epithelial cells in vitro. However, EGF also induced late Akt phosphorylation (after 12 h). Chemical inhibition of PI3K-Akt by LY294002/wortmannin did not affect EGF-mediated NF-κB p65 nuclear translocation; and NF-κB inhibition by Bay 11-7082 did not suppress Akt phosphorylation. Our data suggest that EGF stimulates both the PI3K-Akt pathway and NF-κB via distinct mechanisms, promoting tropic effects in SS salivary epithelial cells.

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Abbreviations

APS:

Aminopropyltriethoxysilane

DAB:

3.3′-diaminobenzidine

DR4:

Death receptor 4

ECL:

Enhanced chemiluminescence

EGF:

Epidermal growth factor

EGF-R:

EGF receptor

FITC:

Fluorescein isothiocyanate

IKK:

IkappaB kinase

NF-κB:

Nuclear factor kappa B

NGF:

Nerve growth factor

PBS:

Phosphate buffered saline

PDGF:

Platelet-derived growth factor

PFA:

Paraformaldehyde

PI3K:

Phosphatidylinositol 3-kinase

PMSF:

Polymethylsulfonylfluoride

PVDF:

Polyvinylidene fluoride

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SS:

Sjögren’s syndrome

TBS:

Tris-buffered saline

TNF:

Tumor necrosis factor

TRITC:

Tetramethyl rhodamine isothiocyanate

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Acknowledgments

The authors would like to thank Dr. Noriyoshi Ogawa from the Third Department of Internal Medicine, Hamamatsu Medical University, for advice regarding the primary cell culture of epithelial cells obtained from minor salivary gland biopsy. We would also like to thank Dr. Yoshitaka Hishikawa from the Division of Histology and Cell Biology, Department of Developmental and Reconstructive Medicine, Graduate School of Biomedical Sciences, Nagasaki University, for his kind advice and technical instruction of pixel intensity measurement.

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Authors and Affiliations

  1. The First Department of Internal Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki City, Nagasaki, 852-8501, Japan

    Hideki Nakamura, Atsushi Kawakami, Hiroaki Ida & Katsumi Eguchi

  2. Division of Histology and Cell Biology, Department of Developmental and Reconstructive Medicine, Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki City, Nagasaki, Japan

    Takehiko Koji

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  1. Hideki Nakamura
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  2. Atsushi Kawakami
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  5. Katsumi Eguchi
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Correspondence to Hideki Nakamura.

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Nakamura, H., Kawakami, A., Ida, H. et al. EGF activates PI3K-Akt and NF-κB via distinct pathways in salivary epithelial cells in Sjögren’s syndrome. Rheumatol Int 28, 127–136 (2007). https://doi.org/10.1007/s00296-007-0411-9

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  • DOI: https://doi.org/10.1007/s00296-007-0411-9

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