Adropin is a multifunctional peptide hormone encoded by the ENHO (energy homeostasis associated) gene. It plays a role in mechanisms related to increased adiposity, insulin resistance, as well as glucose and lipid metabolism. The low adropin levels are strongly associated with obesity independent insulin resistance. On the other hand, overexpression or exogenous administration of adropin improves glucose homeostasis. The multidirectional, adropin-related effects associated with the regulation of metabolism in humans also appear to be attributable to the effects of this peptide on the activity of various elements of the endocrine system including adrenal cortex. Therefore, the main purpose of the present study was to investigate the effect of adropin on proliferation and secretory activity in the human HAC15 adrenal carcinoma cell line.
We also found that HAC15 cells treated with adropin presented significantly higher proliferation levels than untreated cells. Based on whole transcriptome study and research involving transforming growth factor (TGF)-β type I receptor kinase inhibitor we demonstrated that attenuation of steroidogenesis caused by adropin is mediated by the TGF-β signalling pathway likely to act through transactivation mechanism.
Overall design: HAC15 cell line (ATCC® CRL-3301TM, VA, USA) was cultured in a defined medium consisting of DMEM/F-12 without phenol red (Thermo Fisher Scientific, Waltham, MA, USA), 10% Cosmic Calf Serum (Hyclone, GE Healthcare, MA USA), 1% ITS + Premix (Corning, NY, USA), and 1% P/S (Merck Millipore, Darmstadt, Germany). The cells at low passages (p0-p1) underwent starvation in the DMEM/F-12 medium supplemented with 10% charcoal-stripped fetal bovine serum (FBS) (Thermo Fisher Scientific, MA, USA) and 1% P/S for 24h. Subsequently, the cells were treated with the following compounds: ACTH (Synacthen) 10-7 M (Basel, Switzerland), forskolin 25μM (Merck Millipore, Germany), and adropin 10-8 M (Bachem, Switzerland) for another 24h. The cells cultured in the medium with 10% charcoal-stripped FBS served as controls. Immediately after 24h of incubation, the cells and culture media were collected and stored at -80C for microarray study
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