Human NCI-H295 adrenocortical carcinoma cells: A model for angiotensin-II-responsive aldosterone secretion

Ian M. Bird, Neil A. Hanley, R. Ann Word, James Michael Mathis, John L. McCarthy, J. Ian Mason, William E. Rainey

Research output: Contribution to journalArticlepeer-review

152 Scopus citations

Abstract

Excessive secretion of aldosterone from the adrenal results in the most common form of endocrine hypertension. An understanding of the regulatory processes involved in aldosterone synthesis and release is needed to define the biomolecular mechanisms controlling excessive production of aldosterone. However, in vitro studies regarding the regulatory mechanisms of human aldosterone production have been limited because of difficulties in obtaining tissue and the subsequent isolation of aldosterone-secreting glomerulosa cells. Herein we describe an adrenocortical carcinoma cell line, NCI-H295, which provides a suitable angiotensin-II (AH)-responsive model system to investigate the acute and chronic regulation of aldosterone synthesis. The cells were characterized with regard to the effects of All on second messenger systems, aldosterone release, and levels of aldosterone synthase (P450cl8) mRNA. In the presence of lithium, All caused a rapid, but transient, increase in the production of inositol tris-and bisphosphates, whereas a prolonged gradual accumulation of inositol monophosphate occurred. Treatment with All resulted in a 4.5-fold increase in total inositol phosphates in a concentration-dependent manner and an increase in intracellular cytoplasmic free Ca2+. Significant increases in aldosterone (3.5-fold) were detected within 1 h of All addition. Aldosterone release occurred in a concentration-and time-dependent manner. The type 1 All (ATI) receptor was shown to be responsible for activation of phosphoinositidase-C, increased intracellular free Ca2+, and aldosterone production, as determined by use of the ATI receptor antagonist DuP753. In addition, All treatment resulted in a time-dependent increase in levels of P450cl8 mRNA, as detected by RNAse protection assay. In summary, NCI-H295 cells provide a valuable model system to define mechanisms regulating human aldosterone production.

Original languageEnglish
Pages (from-to)1555-1561
Number of pages7
JournalEndocrinology
Volume133
Issue number4
DOIs
StatePublished - 1 Jan 1993

Fingerprint Dive into the research topics of 'Human NCI-H295 adrenocortical carcinoma cells: A model for angiotensin-II-responsive aldosterone secretion'. Together they form a unique fingerprint.

Cite this