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Essential role of Pyk2 and Src kinase activation in neuropeptide-induced proliferation of small cell lung cancer cells

Oncogene volume 27, pages 1737–1748 (2008)Cite this article

Abstract

Neuropeptide hormones like bombesin/gastrin-releasing peptide, galanin or bradykinin, acting via auto and paracrine growth loops, represent the principal mitogens of small cell lung cancer (SCLC). These mitogenic neuropeptides activate Gq/11-coupled receptors which stimulate phospholipase Cβ activity, followed by rises of the intracellular calcium concentration ([Ca2+]i) and activation of protein kinase C (PKC). We report here that proline-rich tyrosine kinase Pyk2 is highly expressed in SCLC cells and provides a functional link between neuropeptide-induced increases in [Ca2+]i and tumor cell proliferation. Activation of Pyk2 and its association with Src kinases critically depends on the elevation of [Ca2+]i, but is independent of PKC. Src kinase activities are crucial for neuropeptide-mediated GTP-loading of Ras and activation of extracellular signal-regulated kinases in SCLC cells. Pyk2 and Src kinases essentially contribute to anchorage-independent proliferation of SCLC cells. Inhibition of either Pyk2 or Src kinases by lentiviral RNAi or pharmacological inhibition with PP2, respectively, attenuated basal and neuropeptide-elicited survival and proliferation of SCLC cells in liquid culture and in soft agar. Thus, neuropeptides stimulate anchorage-independent survival and proliferation of SCLC cells via pathways involving Pyk2 and Src kinases. Therefore, Ca2+-induced Pyk2/Src complex formation may be a rewarding molecular target for novel therapeutic strategies in SCLC cells.

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Abbreviations

ERK:

extracellular signal-regulated kinase

FAK:

focal adhesion kinase

GEF:

guanine nucleotide exchange factor

Grb2:

growth factor receptor-bound protein 2

GRP:

bombesin/gastrin-releasing peptide

JNK/SAPK:

c-jun N-terminal kinase/stress-activated kinase

MAPK:

mitogen-activated protein kinase

MEK:

MAPK/ERK kinase

N-SCLC:

Non-small cell lung cancer

PI3K:

phosphatidylinositol 3-kinase

PKC:

protein kinase C

PLC:

phospholipase C

RasGRF:

Ras guanine nucleotide-releasing factor

RasGRP:

Ras guanine nucleotide-releasing protein

RBD:

Ras-binding domain

SCLC:

small cell lung cancer

SH2 and SH3:

Src homology domain

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Acknowledgements

The study was supported by a grant from the Deutsche Forschungsgemeinschaft.

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Authors and Affiliations

  1. Institut für Pharmakologie und Toxikologie, Philipps-Universität Marburg, Marburg, Germany

    S Roelle, T Buech, V Chubanov & T Gudermann

  2. Pharmakologisches Institut, Ruprecht-Karls-Universität Heidelberg, Heidelberg, Germany

    R Grosse

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  1. S Roelle
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Correspondence to T Gudermann.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Roelle, S., Grosse, R., Buech, T. et al. Essential role of Pyk2 and Src kinase activation in neuropeptide-induced proliferation of small cell lung cancer cells. Oncogene 27, 1737–1748 (2008). https://doi.org/10.1038/sj.onc.1210819

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