Preliminary studies on the differential expression of adrenocorticotropic hormone receptor in adrenal adenomas
Abstract
Introduction: The ACTH receptor (ACTHR) is primarily expressed in the adrenal cortex. Previous studies focused on the regulatory function of ACTHR in glucocorticoid secretion, but research on adrenal tumours is rare. The aim of this study was to evaluate ACTHR expression in common adrenal adenomas and investigate its influence on adrenal tumorigenesis using adrenocortical H295R cells.
Materials and methods: Real-time polymerase chain reaction and western blot were used to detect the expression of ACTHR in 18 aldosterone-producing adenomas, 16 cortisol-producing adenomas, 9 non-functional adenomas, and 12 normal adrenal samples. Lentiviral vector pLVX-mCMV-ACTHR-ZsGreen was transfected into the H295R cells to increase ACTHR expression. WST-1 and cell count were applied to evaluate cell proliferation at different ACTHR levels. TUNEL staining was used to measure cell apoptosis.
Results: Compared with normal adrenal samples, the aldosterone-producing adenoma samples had higher ACTHR mRNA and protein levels. However, the mRNA and protein levels of ACTHR in non-functional adenomas and cortisol-producing adenomas were lower than those in the normal adrenal samples. Proliferative activity in the experimental cells was higher than that in the control cells in the first three days. The proliferative activity peaked in the second day. However, this trend was reversed in the fourth day and became more apparent with time. By contrast, TUNEL staining showed that ACTHR overexpression did not induce a significant difference between the two groups.
Conclusions: Differential ACTHR expression may be determined by different types of adrenocortical tumours. ACTHR is more likely to have induced the proliferation rather than the apoptosis of H295R cells.
Keywords: adrenocorticotropic hormone receptorAdrenal tumourproliferationH295R cells
References
- Mountjoy KG, Robbins LS, Mortrud MT, et al. The cloning of a family of genes that encode the melanocortin receptors. Science. 1992; 257(5074): 1248–1251.
- Reincke M, Beuschlein F, Latronico AC, et al. Expression of adrenocorticotrophic hormone receptor mRNA in human adrenocortical neoplasms: correlation with P450scc expression. Clin Endocrinol. 1997; 46(5): 619–626.
- Chan LiF, Metherell LA, Clark AJL. Effects of melanocortins on adrenal gland physiology. Eur J Pharmacol. 2011; 660(1): 171–180.
- Cao CX, Yang XC, Gao YX, et al. Expression of aldosterone synthase and adrenocorticotropic hormone receptor in adrenal incidentalomas from normotensive and hypertensive patients: Distinguishing subclinical or atypical primary aldosteronism from adrenal incidentaloma. Int J Mol Med. 2012; 30(6): 1396–1402.
- Morris DG, Kola B, Borboli N, et al. Identification of adrenocorticotropin receptor messenger ribonucleic acid in the human pituitary and its loss of expression in pituitary adenomas. J Clin Endocrinol Metab. 2003; 88(12): 6080–6087.
- Hornsby PJ. Aging of the Human Adrenal Cortex. Ageing Res Rev. 2002; 1: 229–242.
- Payet N, Lehoux JG, Isler H. Effect of ACTH on the proliferative and secretory activities of the adrenal glomerulosa. Acta endocrinol (Copenh). 1980; 93(3): 365–374.
- Mazzuco TL, Grunenwald S, Lampron A, et al. Aberrant hormone receptors in primary aldosteronism. Horm Metab Res. 2010; 42(6): 416–423.
- Lenzini L, Rossitto G, Maiolino G, et al. A Meta-Analysis of Somatic KCNJ5 K+Channel Mutations In 1636 Patients With an Aldosterone-Producing Adenoma. J Clin Endocrinol Metab. 2015; 100(8): E1089–E1095.
- Zilbermint M, Stratakis CA. Protein kinase A defects and cortisol-producing adrenal tumors. Curr Opin Endocrinol Diabetes Obes. 2015; 22(3): 157–162.
- Schimmer B, White P. Minireview: Steroidogenic Factor 1: Its Roles in Differentiation, Development, and Disease. Mol Endocrinol. 2010; 24(7): 1322–1337.
- Dongliang Hu, Jinzhi O, Zhun Wu, et al. Elementary studies on elevated steroidogenic factor-1 expression in aldosterone-producing adenoma. Urol Oncol. 2012; 30(4): 457–462.
- Weber A, Toppari J, Harvey RD, et al. Adrenocorticotropin receptor gene mutations in familial glucocorticoid deficiency: relationships with clinical features in four families. J Clin Endocrinol Metab. 1995; 80(1): 65–71.
- Otis M, Campbell S, Payet MD, et al. Expression of extracellular matrix proteins and integrins in rat adrenal gland: importance for ACTH-associated functions. J Endocrinol. 2007; 193(3): 331–347.
- Mountjoy K, Bird I, Rainey W, et al. ACTH induces up-regulation of ACTH receptor mRNA in mouse and human adrenocortical cell lines. Mol Cell Endocrinol. 1994; 99(1): 17–20.
- Webb T, Clark A. Minireview: The Melanocortin 2 Receptor Accessory Proteins. Mol Endocrinol. 2010; 24(3): 475–784.
- Beuschlein F, Fassnacht M, Klink A, et al. ACTH-receptor expression, regulation and role in adrenocortial tumor formation. Eur J Endocrinol. 2001; 144(3): 199–206.
- Reincke M, Beuschlein F, Menig G, et al. Localization and expression of adrenocorticotropic hormone receptor mRNA in normal and neoplastic human adrenal cortex. J Endocrinol. 1998; 156(3): 415–423.
- Ragazzon B, Assie G, Bertherat J. Transcriptome analysis of adrenocortical cancers (ACC): from molecular classification to the identification of new treatments. Endocr Relat Cancer. 2011; 18: 15–27.
- Stojadinovic A, Ghossein RA, Hoos A, et al. Adrenocortical carcinoma: clinical, morphologic, and molecular characterization. J Clin Oncol. 2002; 20(4): 941–950.
- Ilvesmäki V, Kahri AI, Miettinen PJ, et al. Insulin-like growth factors (IGFs) and their receptors in adrenal tumors: high IGF-II expression in functional adrenocortical carcinomas. J Clin Endocrinol Metab. 1993; 77(3): 852–858.
- Arola J, Heikkilä P, Voutilainen R, et al. Biphasic effect of ACTH on growth of rat adrenocortical cells in primary culture. Cell Tissue Res. 1993; 271(1): 169–176.
- Xing Y, Parker CR, Edwards M, et al. ACTH is a potent regulator of gene expression in human adrenal cells. J Mol Endocrinol. 2010; 45(1): 59–68.
![](https://journals.viamedica.pl/plugins/generic/popups/images/icons/close.png)