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Epidermal growth factor receptor mutations in non-small cell lung cancer influence downstream Akt, MAPK and Stat3 signaling

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Abstract

Purpose

The efficacy of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors in non-small cell lung cancer (NSCLC) has been linked to activating mutations in the EGFR gene. So far these mutations have been extensively characterized in established cell lines. The aim of this study was to determine the effects of EGFR mutations on downstream signaling in human tumor specimens.

Methods

We have looked for mutations of the EGFR gene in specimens of 67 patients with NSCLC and correlated these with EGFR phosphorylation and the activity of its three main downstream signaling cascades Akt, MAPK and Stat3 by immunohistochemistry.

Results

We show that the phosphorylation of tyrosine residues 922 and 1173, but not 1068, are primarily affected by the activating EGFR mutations. Akt activity was significantly higher in patients with EGFR mutations but we found no difference in Stat3 or MAPK phosphorylation. Our results suggest that EGFR mutations not only increase receptor activity, but also alter responses of downstream signaling cascades in human NSCLCs and that these finding differ from results obtained in cell lines.

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Abbreviations

EGFR:

Epidermal growth factor receptor

NSCLC:

Non-small cell lung cancer

Akt:

Protein kinase B

MAPK:

Mitogen activated protein kinase

Stat:

Signal transducer and activator of transcription protein

TKI:

Tyrosine kinase inhibitors

PKC:

Protein kinase C

PLCγ:

Phospholipase C-gamma

DAG:

1,2-Diacylglycerol

IP3 :

Inositol 1,3,5-triphosphate

Grb:

Growth factor receptor-bound protein

Sos:

Son of sevenless

SH2:

Src homology 2

GDP/GTP:

Guanosine di-/tri-phosphate

Erk:

Extracellular signal-regulated kinases

PI3-K Ia:

Phosphatidylinositol-3-kinase class Ia

WHO:

World Health Organization

IHC:

Immunohistochemistry

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Acknowledgments

We thank Ellen Paggen for technical help with DNA sequencing and Christiane Esch for support with the immunohistochemistry. L.C.H. was supported by the BONFOR research council.

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Authors and Affiliations

  1. Institute of Pathology, University Hospital Bonn, Sigmund-Freud-Strasse 25, 53127, Bonn, Germany

    Sebastian Zimmer, Philip Kahl, Theresa M. Buhl, Susanne Steiner, Eva Wardelmann, Sabine Merkelbach-Bruse, Reinhard Buettner & Lukas C. Heukamp

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  1. Sebastian Zimmer
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  2. Philip Kahl
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Correspondence to Lukas C. Heukamp.

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Figure 6

Percent of tumors staining positive in EGFR, P-Tyr-992, P-Tyr-1068, P-Tyr-1173, P-Stat3, P-Akt and P-MAPK immunohistochemistry with regard to specific EGFR mutations. The EGFR subgroups were compared to EGFR-WT samples and the P-values are illustrated. The EGFR-M group and the EGFR-M subgroups display the same pattern in receptor phosphorylation and activity of downstream signaling cascades. (EPS 438 kb)

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Zimmer, S., Kahl, P., Buhl, T.M. et al. Epidermal growth factor receptor mutations in non-small cell lung cancer influence downstream Akt, MAPK and Stat3 signaling. J Cancer Res Clin Oncol 135, 723–730 (2009). https://doi.org/10.1007/s00432-008-0509-9

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  • DOI: https://doi.org/10.1007/s00432-008-0509-9

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