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Ligand release-independent transactivation of epidermal growth factor receptor by transforming growth factor-β involves multiple signaling pathways

Oncogene volume 27, pages 614–628 (2008)Cite this article

Abstract

Many of the signaling responses induced by transforming growth factor-β (TGF-β) are mediated by Smad proteins, but there is evidence that it can also signal independently of Smads. Here, we provide evidence that multiple signal pathways induced by TGF-β1—including Src family tyrosine kinases (SFKs), generation of reactive oxygen species (ROS), de novo protein synthesis and E-cadherin-dependent cell–cell interactions—transactivate the epidermal growth factor receptor (EGFR), which in turn regulates expression of c-Fos and c-Jun. Immunoprecipitation and immunofluorescence staining showed that EGFR was phosphorylated on tyrosine in response to TGF-β1. EGFR transactivation required the activation of SFKs and the production of ROS via NADPH oxidase, but was not dependent on metalloproteases or the release of EGF-like ligands. In addition, the production of ROS was dependent on signaling by specific SFKs as well as de novo protein synthesis. Stable transfection of E-cadherin into MDA-MB-231 cells as well as E-cadherin-blocking assays revealed that E-cadherin-mediated cell–cell interactions were also essential for EGFR transactivation. Finally, EGFR transactivation was involved in the expression of c-Fos and c-Jun via the extracellular signal-regulated kinase signaling cascade. Taken together our data suggest that ligand release-independent transactivation of EGFR may diversify early TGF-β signaling and represent a novel pathway leading to TGF-β-mediated gene expression.

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Abbreviations

ADAM:

a disintegrin and a metalloproteinase

BMP:

bone morphogenetic protein

BSA:

bovine serum albumin

DMEM:

Dulbecco's modified Eagle's medium

DPI:

diphenylene iodonium

EGFR:

epidermal growth factor receptor

ELISA:

enzyme-linked immunosorbent assay

EMT:

epithelial to mesenchymal transition

ERK:

extracellular signal-regulated kinase

FBS:

fetal bovine serum

GPCR:

G-protein-coupled receptor

HB-EGF:

heparin-binding-epidermal growth factor

JNK:

c-Jun N-terminal kinase

LC-MS/MS:

liquid chromatography-mass spectrometry/mass spectrometry

mAb:

monoclonal antibody

MAPK:

mitogen-activated protein kinase

MET:

mesenchymal to epithelial transition

MMP:

metalloproteinase

NAC:

N-acetyl cysteine

pAb:

polyclonal antibody

PBS:

phosphate-buffered saline

PTP:

protein-tyrosine phosphatases

ROS:

reactive oxygen species

SDS–PAGE:

sodium dodecylsulfate polyacrylamide gel electrophoresis

SFK:

Src family tyrosine kinase

TGF:

transforming growth factor

TMPS:

triple-membrane-passing signaling

TPA:

12-O-tetradecanoylphorbol-13-acetate

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Acknowledgements

We thank Dr Eunjoo H Lee for her invaluable comments and critical review of this manuscript; Dr P Cossart (Institute of Pasteur, France) for providing human E-cadherin plasmid. This work was supported by Korea Research Foundation Grant (KRF-2004-041-C00307).

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Authors and Affiliations

  1. Laboratory of Visual Science, Korea Eye Tissue and Gene Bank, College of Medicine, The Catholic University of Korea, Seoul, Korea

    C-K Joo, H-S Kim, J-Y Park, Y Seomun, M-J Son & J-T Kim

  2. Bio Division, KOCAT Inc., Gasan-Dong, Geumcheon-Ku, Seoul, Korea

    J-T Kim

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Joo, CK., Kim, HS., Park, JY. et al. Ligand release-independent transactivation of epidermal growth factor receptor by transforming growth factor-β involves multiple signaling pathways. Oncogene 27, 614–628 (2008). https://doi.org/10.1038/sj.onc.1210649

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