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J. Biol. Chem., Vol. 276, Issue 47, 43668-43676, November 23, 2001

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Proliferative Actions of Natriuretic Peptides on Neuroblastoma Cells
INVOLVEMENT OF GUANYLYL CYCLASE AND NON-GUANYLYL CYCLASE PATHWAYS^*

Vincent Lelièvre, Nicolas Pineau^§, Zhongting Hu, Yevgeniya Ioffe, Ji-Yun Byun, Jean-Marc Muller^§, and James A. Waschek^¶

From the  Department of Psychiatry and Mental Retardation Research Center, UCLA, Neuropsychiatric Institute, Los Angeles, California 90024 and ^§ CNRS UMR 6558, Laboratoire Biologie des Interactions Cellulaires, UFR Sciences, Université de Poitiers, 40 avenue du Recteur PINEAU, 86022 Poitiers Cedex, France

To identify neural tumor cell lines that could be used as models to study growth-related natriuretic peptide actions, we determined the effects of these peptides on the proliferation of human and rodent neuroblastoma cell lines. Subnanomolar concentrations of atrial natriuretic peptide (ANP) and type C natriuretic peptide (CNP) stimulated proliferation in all four cell lines. These actions were associated with cGMP elevation and were blocked by a protein kinase G inhibitor. These data imply the involvement of guanylyl cyclase (GC)-coupled natriuretic receptors. However, higher concentrations of ANP and CNP, and low concentrations of des-[Gln¹⁸,Ser¹⁹,Gly²⁰,Leu²¹,Gly²²]-ANP_4-23-NH₂ (desANP_4-23) (analog for NPR-C receptor) exerted antiproliferative actions in three of the cell lines. These effects were insensitive to a protein kinase G inhibitor and to HS-142-1, suggesting that growth-inhibitory actions involved a non-GC receptor. They did not appear to involve cAMP, protein kinase A, protein kinase C, or calcium mobilization but were abolished when constitutive mitogen-activated protein kinase activity was inhibited. Radioligand binding experiments revealed the presence of a uniform class of binding sites in NG108 cells and multiple binding sites in Neuro2a cells. Northern and reverse transcriptase-polymerase chain reaction analyses revealed differential gene expression for NPR-A/B/C in NG108 and Neuro2a cells. The results indicate that natriuretic peptides stimulate neuroblastoma cell proliferation through type NPR-A/B (GC) receptors. Higher concentrations of ANP and CNP exerted a mitogen-activated protein kinase-dependent antiproliferative action mediated by a non-GC receptor that interacts with desANP_4-23 with relatively high affinity.

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