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Pituitary Adenylate Cyclase-Activating Polypeptide Causes Increased Tyrosine Phosphorylation of Focal Adhesion Kinase and Paxillin

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Abstract

The effects of pituitary adenylate cyclase-activating polypeptide (PACAP) on tyrosine phosphorylation of focal adhesion kinase (FAK) and paxillin were investigated using lung cancer cells. Addition of PACAP-27 or PACAP-38 but not vasoactive intestinal peptide to NCI-H838 or NCI-H1299 human lung cancer cells significantly increased the tyrosine phosphorylation of FAK or paxillin. The increase in FAK or paxillin tyrosine phosphorylation caused by addition of PACAP-27 to NCI-H838 cells was inhibited by PACAP(6–38), a PAC1-receptor (R) antagonist. The increase in FAK or paxillin tyrosine phosphorylation caused by 100 nM PACAP-27 was maximal 2 min after addition to NCI-H838 cells. The effects of PACAP at stimulating FAK and paxillin tyrosine phosphorylation were reversed by cytochalasin D and genistein which inhibit actin polymerization and tyrosine kinase activity, respectively. The effects of PACAP at stimulating FAK and paxillin tyrosine phosphorylation were reversed by U-73122 but not H89 which inhibit phospholipase C and protein kinase A, respectively. The results show that PAC1-R regulates FAK and paxillin tyrosine phosphorylation in lung cancer cells as a result of increased phosphatidylinositol turnover but not adenylyl cylase stimulation.

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Abbreviations

PACAP:

Pituitary adenylate cyclase-activating polypeptide

FAK:

Focal adhesion kinase

R:

Receptor

SCLC:

Small cell lung cancer

NSCLC:

Non-SCLC

VIP:

Vasoactive intestinal peptide

GRP:

Gastrin-releasing peptide

Ca2+ :

Calcium

GPCR:

G-protein-coupled receptor

pY:

Phosphotyrosine

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Acknowledgments

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

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Authors and Affiliations

  1. Center for Cancer Research, National Cancer Institute, 31 Center Dr., Bldg. 31, Rm. 4A48, Bethesda, MD, 20892, USA

    Terry W. Moody

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

    Julius Leyton & Robert T. Jensen

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  1. Terry W. Moody
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  2. Julius Leyton
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Correspondence to Terry W. Moody.

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Moody, T.W., Leyton, J. & Jensen, R.T. Pituitary Adenylate Cyclase-Activating Polypeptide Causes Increased Tyrosine Phosphorylation of Focal Adhesion Kinase and Paxillin. J Mol Neurosci 46, 68–74 (2012). https://doi.org/10.1007/s12031-011-9639-7

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