AbstractIntroduction. Human ghrelin is the endogenous ligand of the growth hormone secretagogue receptor type 1a (GHSR1a). It is suggested that ghrelin is involved in pituitary adenomas pathogenesis. There are inconsistent data regarding the effect of ghrelin on cell proliferation. In this study the outcome of ghrelin in the rat pituitary adenoma GH3 cell line on morphology and proliferation ratio was evaluated. The ghrelin receptor (Ghsr) mRNA expression in GH3 cell line was established as well, because it was found that heterogeneous expression pattern characterized physiological and pathological conditions of tissues of different origin.
Material and Methods. Suitable experimental model pituitary tumor (rat GH3 cell line) was stimulated with ghrelin in the final concentrations 10–12 M, 10–9 M and 10–6 M. Reverse transcription followed by real time polymerase chain reaction was used for ghrelin receptor gene transcript detection. The morphology as well as cell cycle of those cells were analyzed using Axio Vert.A1 Microscope (Zeiss) and BD FACSCalibur™ flow cytometer (Beckton Dickinson), respectively. The percentages of cells in the G0/G1, S, G2/M cycle phases were evaluated using the ModFit™ software (Verity Software, Inc., USA).
Results. Ghsr mRNA presence was confirmed in GH3 cells. Ghrelin did not affect conspicuously GH3 cells morphology, however the ghrelin-induced proliferation index increase was caused by both decline of G0/G1 phases cells count and increase those being in S+G2/M (p < 0.05).
Conclusions. In conclusion, this study indicates that ghrelin stimulates GH3 cells proliferation and may play role in pituitary tumorigenesis via an autocrine/paracrine pathway.
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- Arvat E, Di Vito L, Broglio F, Papotti M, Muccioli G, Dieguez C et al. Preliminary evidence that Ghrelin, the natural GH secretagogue (GHS)-receptor ligand, strongly stimulates GH secretion in humans. J Endocrinol Invest. 2000 Sep;23(8):493–5.
- Cassoni P, Papotti M, Ghe C, Catapano F, Sapino A, Graziani A et al. Identification, characterization, and biological activity of specific receptors for natural (ghrelin) and synthetic growth hormone secretagogues and analogs in human breast carcinomas and cell lines. J Clin Endocrinol Metab. 2001 Apr;86(4):1738–45.
- Leite-Moreira AF, Soares J-B. Physiological, pathological and potential therapeutic roles of ghrelin. Drug Discov Today. 2007 Apr;12(7–8):276–88.
- Murata M, Okimura Y, Iida K, Matsumoto M, Sowa H, Kaji H et al. Ghrelin modulates the downstream molecules of insulin signaling in hepatoma cells. J Biol Chem. 2002 Feb 15;277(7):5667–74.
- Nanzer AM, Khalaf S, Mozid AM, Fowkes RC, Patel M V, Burrin JM et al. Ghrelin exerts a proliferative effect on a rat pituitary somatotroph cell line via the mitogen-activated protein kinase pathway. Eur J Endocrinol. 2004 Aug;151(2):233–40.
- Stevanovic D, Milosevic V, Nesic D, Ajdzanovic V, Starcevic V, Severs WB. Central effects of ghrelin on serum growth hormone and morphology of pituitary somatotropes in rats. Exp Biol Med (Maywood). 2006 Nov;231(10):1610–5.
- Milosević VL, Stevanović DM, Nesić DM, Sosić-Jurjević BT, Ajdzanović VZ, Starcević VP et al. Central effects of ghrelin on the adrenal cortex: a morphological and hormonal study. Gen Physiol Biophys. 2010 Jun;29(2):194–202.
- Muccioli G, Ghe C, Ghigo MC, Papotti M, Arvat E, Boghen MF et al. Specific receptors for synthetic GH secretagogues in the human brain and pituitary gland. J Endocrinol. 1998 Apr;157(1):99–106.
- Muccioli G, Papotti M, Locatelli V, Ghigo E, Deghenghi R. Binding of 125I-labeled ghrelin to membranes from human hypothalamus and pituitary gland. J Endocrinol Invest. 2001 Mar;24(3):RC7–9.
- Kim K, Arai K, Sanno N, Osamura RY, Teramoto A, Shibasaki T. Ghrelin and growth hormone (GH) secretagogue receptor (GHSR) mRNA expression in human pituitary adenomas. Clin Endocrinol (Oxf). 2001 Jun;54(6):759–68.
- Jeffery PL, Herington AC, Chopin LK. The potential autocrine/paracrine roles of ghrelin and its receptor in hormone-dependent cancer. Cytokine Growth Factor Rev. 2003 Apr;14(2):113–22.
- Delhanty PJD, van Koetsveld PM, Gauna C, van de Zande B, Vitale G, Hofland LJ et al. Ghrelin and its unacylated isoform stimulate the growth of adrenocortical tumor cells via an anti-apoptotic pathway. Am J Physiol Endocrinol Metab. 2007 Jul;293(1):E302–9.
- Zhang W, Lin TR, Hu Y, Fan Y, Zhao L, Stuenkel EL et al. Ghrelin stimulates neurogenesis in the dorsal motor nucleus of the vagus. J Physiol. 2004 Sep;559(Pt 3):729–37.
- Zhang W, Zhao L, Lin TR, Chai B, Fan Y, Gantz I et al. Inhibition of adipogenesis by ghrelin. Mol Biol Cell. 2004 May;15(5):2484–91.
- Baldanzi G, Filigheddu N, Cutrupi S, Catapano F, Bonissoni S, Fubini A et al. Ghrelin and des-acyl ghrelin inhibit cell death in cardiomyocytes and endothelial cells through ERK1/2 and PI 3-kinase/AKT. J Cell Biol. 2002 Dec;159(6):1029–37.
- Tian C, Ye F, Wang L, Deng Y, Dong Y, Wang X et al. Nitric oxide inhibits ghrelin-induced cell proliferation and ERK1/2 activation in GH3 cells. Endocrine. 2010 Dec;38(3):412–6.
- Gahete MD, Rincón-Fernández D, Villa-Osaba A, Hormaechea-Agulla D, Ibánez-Costa A, Martínez-Fuentes AJ et al. Ghrelin gene products, receptors, and GOAT enzyme: biological and pathophysiological insight. J Endocrinol. 2014 Jan;220(1):R1–24.
- Papotti M, Ghe C, Cassoni P, Catapano F, Deghenghi R, Ghigo E et al. Growth hormone secretagogue binding sites in peripheral human tissues. J Clin Endocrinol Metab. 2000 Oct;85(10):3803–7.
- Volante M, Allia E, Fulcheri E, Cassoni P, Ghigo E, Muccioli G et al. Ghrelin in Fetal Thyroid and Follicular Tumors and Cell Lines. Am J Pathol. 2003 Feb;162(2):645–54.
- Cassoni P, Papotti M, Catapano F, Ghe C, Deghenghi R, Ghigo E et al. Specific binding sites for synthetic growth hormone secretagogues in non-tumoral and neoplastic human thyroid tissue. J Endocrinol. 2000 Apr;165(1):139–46.
- van der Lely AJ, Tschöp M, Heiman ML, Ghigo E. Biological, physiological, pathophysiological, and pharmacological aspects of ghrelin. Endocr Rev. 2004 Jun;25(3):426–57.
- Ghigo E, Broglio F, Arvat E, Maccario M, Papotti M, Muccioli G. Ghrelin: more than a natural GH secretagogue and/or an orexigenic factor. Clin Endocrinol (Oxf). 2005 Jan;62(1):1–17.
- Maccarinelli G, Sibilia V, Torsello A, Raimondo F, Pitto M, Giustina A et al. Ghrelin regulates proliferation and differentiation of osteoblastic cells. J Endocrinol. 2005 Jan 1;184(1):249–56.
- Tannenbaum GS, Epelbaum J, Bowers CY. Interrelationship between the novel peptide ghrelin and somatostatin/growth hormone-releasing hormone in regulation of pulsatile growth hormone secretion. Endocrinology. 2003 Mar;144(3):967–74.