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Direct promoter induction of p19Arf by Pit-1 explains the dependence receptor RET/Pit-1/p53-induced apoptosis in the pituitary somatotroph cells

Oncogene volume 31, pages 2824–2835 (2012)Cite this article

Abstract

Somatotrophs produce growth hormone (GH) and are the most abundant secretory cells of the pituitary. Somatotrophs express the transcription factor Pit-1 and the dependence receptor RET, its co-receptor GFRa1 and ligand GDNF. Pit-1 is a transcription factor essential for somatotroph proliferation and differentiation and for GH expression. GDNF represses excess Pit-1 expression preventing excess GH. In the absence of GDNF, RET behaves as a dependence receptor, becomes intracellularly processed and induces strong Pit-1 expression leading to p53 accumulation and apoptosis. How accumulation of Pit-1 leads to p53 expression is unknown. We have unveiled the relationship of Pit-1 with the p19Arf gene. There is a parallel correlation of RET processing, Pit-1 increase and ARF protein and mRNA expression. Interfering the pathway with RET, Pit-1 or p19Arf siRNA blocked apoptosis. We have found a Pit-1 DNA-binding element within the ARF promoter. Pit-1 directly regulates the CDKN2A locus and binds to the p19Arft promoter inducing p19Arf gene expression. The Pit-1-binding element is conserved in rodents and humans. RET/Pit-1 induces p19Arf/p53 and apoptosis not only in a somatotroph cell line but also in primary cultures of pituitary somatotrophs, where ARF siRNA interference also blocks p53 and apoptosis. Analyses of the somatotrophs in whole pituitaries supported the above findings. Thus Pit-1, a differentiation factor, activates the oncogene-induced apoptosis (OIA) pathway as oncogenes exerting a tight control in somatotrophs to prevent the disease due to excess of GH (insulin-resistance, metabolic disease, acromegaly).

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Abbreviations

AP:

anterior pituitary

GH:

growth hormone

OIA:

oncogene-induced apoptosis

RET-S or RET-L:

RET tyrosine kinase receptor short isoform or long isoform

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Acknowledgements

We thank Susana Bravo and Maria Garcia-Rendueles for helpful methodological advice. This project has been supported by grants Xunta de Galicia-MICINN-FEDER, 06PXIB208107PR, 09CSA011208PR, BFU2007-60571 and BFU2010-16652 to CVA. E D-R has been an Anxeles Alvariño fellow and M G-L is a Lucas Labrada fellow (Xunta de Galicia). These programs are co-financed by the European Community (Fondo Social Europeo). The text has been reviewed by G Lockhart, a professional editor of biomedical English.

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  1. C Cañibano

    Present address: 4Current address: Ibiomed, Leon, Spain.,

Authors and Affiliations

  1. Department of Physiology, IDIS-Neoplasia and Endocrine Differentiation, CIMUS, School of Medicine, University of Santiago de Compostela USC, Santiago de Compostela, Spain

    E Diaz-Rodriguez, M García-Lavandeira, S Perez-Romero, A Senra, C Cañibano, C Dieguez & C V Alvarez

  2. Instituto de Investigaciones Biomédicas ‘Alberto Sols’ CSIC-UAM, Madrid, Spain

    I Palmero

  3. Department of Experimental Oncology, Operative Unit Molecular Mechanisms of Cancer Growth and Progression, IRCCS Istituto Nazionale dei Tumori Foundation, Milan, Italy

    M G Borrello

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Diaz-Rodriguez, E., García-Lavandeira, M., Perez-Romero, S. et al. Direct promoter induction of p19Arf by Pit-1 explains the dependence receptor RET/Pit-1/p53-induced apoptosis in the pituitary somatotroph cells. Oncogene 31, 2824–2835 (2012). https://doi.org/10.1038/onc.2011.458

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