Comparative proteomic analysis to identify the novel target gene of angiotensin II in adrenocortical H295R cells

Endocr J. 2021 Apr 28;68(4):441-450. doi: 10.1507/endocrj.EJ20-0144. Epub 2020 Dec 26.

Abstract

Angiotensin II (Ang II) is a well-known peptide that maintains the balance of electrolytes in the higher vertebrates. Ang II stimulation in the adrenal gland induces the synthesis of mineralocorticoids, mainly aldosterone, through the up-regulation of aldosterone synthase (CYP11B2) gene expression. Additionally, it has been reported that Ang II activates multiple signaling pathways such as mitogen-activated protein kinase (MAPK) and Ca2+ signaling. Although Ang II has various effects on the cellular signaling in the adrenal cells, its biological significance, except for the aldosterone synthesis, is still unclear. In this study, we attempted to search the novel target gene(s) of Ang II in the human adrenal H295R cells using a proteomic approach combined with stable isotopic labeling using amino acid in cell culture (SILAC). Interestingly, we found that Ang II stimulation elevated the expression of phosphofructokinase type platelet (PFKP) in both protein and mRNA levels. Moreover, transactivation of PFKP by Ang II was dependent on extracellular-signal-regulated kinase (ERK) 1/2 activation. Finally, we observed that Ang II treatment facilitated glucose uptake in the H295R cells. Taken together, we here identified PFKP as a novel target gene of Ang II, indicating that Ang II not only stimulates steroidogenesis but also affects glucose metabolism.

Keywords: Adrenocortical cell; Angiotensin II; Phosphofructokinase type platelet; Proteome analysis.

MeSH terms

  • Adrenal Cortex / drug effects*
  • Adrenal Cortex / metabolism
  • Angiotensin II / pharmacology
  • Cell Line
  • Cytochrome P-450 CYP11B2 / genetics*
  • Cytochrome P-450 CYP11B2 / metabolism
  • Gene Expression / drug effects*
  • Humans
  • Proteomics
  • Signal Transduction / drug effects
  • Up-Regulation / drug effects

Substances

  • Angiotensin II
  • Cytochrome P-450 CYP11B2