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Horm Metab Res 2012; 44(03): 245-250
DOI: 10.1055/s-0031-1298019
Original Basic
© Georg Thieme Verlag KG Stuttgart · New York

Comparison of Aldosterone Production Among Human Adrenocortical Cell Lines

T. Wang
1   Department of Physiology, Georgia Health Sciences University, Augusta, GA, USA
2   Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan, P. R. China
,
J. G. Rowland
1   Department of Physiology, Georgia Health Sciences University, Augusta, GA, USA
,
J. Parmar
1   Department of Physiology, Georgia Health Sciences University, Augusta, GA, USA
,
M. Nesterova
3   Section on Endocrinology Genetics, Eunice Kennedy Shriver National Institute of Child Health & Human Development, National Institutes of Health Clinical Center, NIH, Bethesda, MD, USA
,
T. Seki
1   Department of Physiology, Georgia Health Sciences University, Augusta, GA, USA
,
W. E. Rainey
1   Department of Physiology, Georgia Health Sciences University, Augusta, GA, USA
› Author Affiliations
Further Information

Publication History

received 30 August 2011

accepted 28 November 2011

Publication Date:
20 January 2012 (online)

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Abstract

Several human adrenocortical cell lines have been used as model systems for aldosterone production. However, these cell lines have not been directly compared with each other. Human adrenal cell lines SW13, CAR47, the NCI-H295 and its sub-strains and sub-clones were compared with regard to aldosterone production and aldosterone synthase (CYP11B2) expression. Culture media was collected 48 h after incubation, aldosterone secretion was measured and the data were normalized to the amount of cell protein. RNA was isolated for microarray analysis and quantitative RT-PCR (qPCR). The cell lines with the highest aldosterone production were further tested with regard to angiotensin II (Ang II) stimulation. Neither aldosterone nor CYP11B2 transcript were detected in SW13 or CAR47 cells. The aldosterone production by the NCI-H295, H295A, H295R-S1, H295R-S2, H295R-S3, HAC13, HAC15 and HAC50 were 119, 1, 6, 826, 18, 139, 412, and 1 334 (pmol/mg protein/48 h), respectively. H295A and H295R-S1 expressed less CYP11B2 than the commonly used H295R-S3 cells; while NCI-H295, H295R-S2, HAC13, HAC15 and HAC50 expressed 24-, 14-, 3-, 10-, and 35-fold higher CYP11B2 compared with the H295R-S3 cells. When treated with Ang II, NCI-H295, H295R-S2, HAC13, HAC15 and HAC50 showed significantly higher aldosterone production than the basal level (p<0.05). A comparison of the available human adrenal cell lines indicates that the H295R-S2 and the clonal cell lines, HAC13, HAC15 and HAC50 produced the highest levels of aldosterone and responded well to Ang II.

Key words

aldosterone - adrenocortical carcinoma - steroidogenesis
 

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