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PYK-2 is Tyrosine Phosphorylated after Activation of Pituitary Adenylate Cyclase Activating Polypeptide Receptors in Lung Cancer Cells

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Abstract

The signal transduction mechanisms of pituitary adenylate cyclase activating polypeptide (PACAP) were investigated in lung cancer cells. Previously, PACAP-27 addition to NCI-H838 cells increased phosphatidylinositol turnover and intracellular cAMP leading to proliferation of lung cancer cells. Also, PACAP receptors (PAC1) regulated the tyrosine phosphorylation of ERK, focal adhesion kinase, and paxillin. In this communication, the effects of PACAP on cytosolic Ca2+ and PYK-2 tyrosine phosphorylation were investigated. PACAP-27 increased cytosolic Ca2+ within seconds after addition to FURA-2 AM loaded NCI-H838 cells. The increase in cytosolic Ca2+ caused by PACAP was inhibited by PACAP(6–38) (PAC1 antagonist), U73122 (phospholipase C inhibitor), or BAPTA (calcium chelator), but not H89 (PKA inhibitor). PACAP-38, but not vasoactive intestinal peptide (VIP), addition to NCI-H838 or H1299 cells significantly increased the tyrosine phosphorylation of PYK-2 after 2 min. The increase in PYK-2 tyrosine phosphorylation caused by PACAP was inhibited by PACAP(6–38), U73122, or BAPTA, but not H89. The results suggest that PAC1 regulates PYK-2 tyrosine phosphorylation in a calcium-dependent manner.

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Abbreviations

PACAP:

Pituitary adenylate cyclase activating polypeptide

PYK-2:

Proline-rich tyrosine kinase 2

FAK:

Focal adhesion kinase

R:

Receptor

SCLC:

Small cell lung cancer

NSCLC:

Non-SCLC

VIP:

Vasoactive intestinal peptide

Ca2+ :

Calcium

GPCR:

G-protein coupled receptor

pY:

Phosphotyrosine

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Acknowledgment

This research is supported in part by intramural funds of the NCI and NIDDK of NIH.

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Authors and Affiliations

  1. NCI Office of the Director, Center for Cancer Research, National Cancer Institute, 31 Center Dr., Bldg. 31, Rm. 4A48, Bethesda, MD, 20892, USA

    Terry W. Moody

  2. Digestive Diseases Branch, National Institutes of Diabetes and Digestive and Kidney Disease, National Institutes of Health, Bethesda, MD, 20892, USA

    Alessia Di Florio & Robert T. Jensen

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  1. Terry W. Moody
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  2. Alessia Di Florio
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  3. Robert T. Jensen
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Correspondence to Terry W. Moody.

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Moody, T.W., Di Florio, A. & Jensen, R.T. PYK-2 is Tyrosine Phosphorylated after Activation of Pituitary Adenylate Cyclase Activating Polypeptide Receptors in Lung Cancer Cells. J Mol Neurosci 48, 660–666 (2012). https://doi.org/10.1007/s12031-012-9785-6

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