Download PDF
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)

Also available at
    Permissions and Reprints

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
 

  • References

  • 1 Hattangady NG, Olala LO, Bollag WB, Rainey WE. Acute and chronic regulation of aldosterone production. Mol Cell Endocrinol 2011. Aug 4. [Epub ahead of print]
  • 2 Clark BJ, Pezzi V, Stocco DM, Rainey WE. The steroidogenic acute regulatory protein is induced by angiotensin II and K+ in H295R adrenocortical cells. Mol Cell Endocrinol 1995; 115: 215-219
    CrossrefPubMedGoogle Scholar
  • 3 Clark BJ, Soo SC, Caron KM, Ikeda Y, Parker KL, Stocco DM. Hormonal and developmental regulation of the steroidogenic acute regulatory protein. Mol Endocrinol 1995; 9: 1346-1355
    CrossrefPubMedGoogle Scholar
  • 4 Clark BJ, Wells J, King SR, Stocco DM. The purification, cloning, and expression of a novel luteinizing hormone-induced mitochondrial protein in MA-10 mouse Leydig tumor cells. Characterization of the steroidogenic acute regulatory protein (StAR). J Biol Chem 1994; 269: 28314-28322
    PubMedGoogle Scholar
  • 5 Kater CE, Biglieri EG, Rost CR, Schambelan M, Hirai J, Chang BC, Brust N. The constant plasma 18-hydroxycorticosterone to aldosterone ratio: an expression of the efficacy of corticosterone methyloxidase type II activity in disorders with variable aldosterone production. J Clin Endocrinol Metab 1985; 60: 225-228
    CrossrefPubMedGoogle Scholar
  • 6 Mornet E, Dupont J, Vitek A, White PC. Characterization of two genes encoding human steroid 11 beta-hydroxylase (P-450(11) beta). J Biol Chem 1989; 264: 20961-20967
    CrossrefPubMedGoogle Scholar
  • 7 Ogishima T, Suzuki H, Hata J, Mitani F, Ishimura Y. Zone-specific expression of aldosterone synthase cytochrome P-450 and cytochrome P-45011 beta in rat adrenal cortex: histochemical basis for the functional zonation. Endocrinology 1992; 130: 2971-2977
    CrossrefPubMedGoogle Scholar
  • 8 Wotus C, Levay-Young BK, Rogers LM, Gomez-Sanchez CE, Engeland WC. Development of adrenal zonation in fetal rats defined by expression of aldosterone synthase and 11beta-hydroxylase. Endocrinology 1998; 139: 4397-4403
    CrossrefPubMedGoogle Scholar
  • 9 Rainey WE. Adrenal zonation: clues from 11beta-hydroxylase and aldosterone synthase. Mol Cell Endocrinol 1999; 151: 151-160
    CrossrefPubMedGoogle Scholar
  • 10 Quinn SJ, Williams GH. Regulation of aldosterone secretion. Annu Rev Physiol 1988; 50: 409-426
    CrossrefPubMedGoogle Scholar
  • 11 Nogueira EF, Bollag WB, Rainey WE. Angiotensin II regulation of adrenocortical gene transcription. Mol Cell Endocrinol 2009; 302: 230-236
    CrossrefPubMedGoogle Scholar
  • 12 Yagci A, Oertle M, Seiler H, Schmid D, Campofranco C, Muller J. Potassium induces multiple steroidogenic enzymes in cultured rat zona glomerulosa cells. Endocrinology 1996; 137: 2406-2414
    CrossrefPubMedGoogle Scholar
  • 13 Szekeres M, Turu G, Orient A, Szalai B, Supeki K, Cserzo M, Varnai P, Hunyady L. Mechanisms of angiotensin II-mediated regulation of aldosterone synthase expression in H295R human adrenocortical and rat adrenal glomerulosa cells. Mol Cell Endocrinol 2009; 302: 244-253
    CrossrefPubMedGoogle Scholar
  • 14 Bassett MH, Mayhew B, Rehman K, White PC, Mantero F, Arnaldi G, Stewart PM, Bujalska I, Rainey WE. Expression profiles for steroidogenic enzymes in adrenocortical disease. J Clin Endocrinol Metab 2005; 90: 5446-5455
    CrossrefPubMedGoogle Scholar
  • 15 Fallo F, Pezzi V, Barzon L, Mulatero P, Veglio F, Sonino N, Mathis JM. Quantitative assessment of CYP11B1 and CYP11B2 expression in aldosterone-producing adenomas. Eur J Endocrinol 2002; 147: 795-802
    CrossrefPubMedGoogle Scholar
  • 16 Ogishima T, Shibata H, Shimada H, Mitani F, Suzuki H, Saruta T, Ishimura Y. Aldosterone synthase cytochrome P-450 expressed in the adrenals of patients with primary aldosteronism. J Biol Chem 1991; 266: 10731-10734
    PubMedGoogle Scholar
  • 17 Gomez-Sanchez CE, Rossi GP, Fallo F, Mannelli M. Progress in primary aldosteronism: present challenges and perspectives. Horm Metab Res 2010; 42: 374-381
    Article in Thieme ConnectPubMedGoogle Scholar
  • 18 Bird IM, Hanley NA, Word RA, Mathis JM, McCarthy JL, Mason JI, Rainey WE. Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin-II-responsive aldosterone secretion. Endocrinology 1993; 133: 1555-1561
    CrossrefPubMedGoogle Scholar
  • 19 Rainey WE, Bird IM, Mason JI. The NCI-H295 cell line: a pluripotent model for human adrenocortical studies. Mol Cell Endocrinol 1994; 100: 45-50
    CrossrefPubMedGoogle Scholar
  • 20 Rainey WE, Saner K, Schimmer BP. Adrenocortical cell lines. Mol Cell Endocrinol 2004; 228: 23-38
    CrossrefPubMedGoogle Scholar
  • 21 Staels B, Hum DW, Miller WL. Regulation of steroidogenesis in NCI-H295 cells: a cellular model of the human fetal adrenal. Mol Endocrinol 1993; 7: 423-433
    CrossrefPubMedGoogle Scholar
  • 22 Nesterova M, Bossis I, Wen F, Horvath A, Matyakhina L, Stratakis CA. An immortalized human cell line bearing a PRKAR1A-inactivating mutation: effects of overexpression of the wild-type Allele and other protein kinase A subunits. J Clin Endocrinol Metab 2008; 93: 565-571
    CrossrefPubMedGoogle Scholar
  • 23 Leibovitz A, McCombs 3rd WM, Johnston D, McCoy CE, Stinson JC. New human cancer cell culture lines. I. SW-13, small-cell carcinoma of the adrenal cortex. J Natl Cancer Inst 1973; 51: 691-697
    PubMedGoogle Scholar
  • 24 Gazdar AF, Oie HK, Shackleton CH, Chen TR, Triche TJ, Myers CE, Chrousos GP, Brennan MF, Stein CA, La Rocca RV. Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. Cancer Res 1990; 50: 5488-5496
    PubMedGoogle Scholar
  • 25 Rodriguez H, Hum DW, Staels B, Miller WL. Transcription of the human genes for cytochrome P450scc and P450c17 is regulated differently in human adrenal NCI-H295 cells than in mouse adrenal Y1 cells. J Clin Endocrinol Metab 1997; 82: 365-371
    CrossrefPubMedGoogle Scholar
  • 26 Xing Y, Edwards MA, Ahlem C, Kennedy M, Cohen A, Gomez-Sanchez CE, Rainey WE. The effects of ACTH on steroid metabolomic profiles in human adrenal cells. J Endocrinol 2011; 209: 327-335
    CrossrefPubMedGoogle Scholar
  • 27 Pezzi V, Mathis JM, Rainey WE, Carr BR. Profiling transcript levels for steroidogenic enzymes in fetal tissues. J Steroid Biochem Mol Biol 2003; 87: 181-189
    CrossrefPubMedGoogle Scholar
  • 28 Wang T, Satoh F, Morimoto R, Nakamura Y, Sasano H, Auchus RJ, Edwards MA, Rainey WE. Gene expression profiles in aldosterone-producing adenomas and adjacent adrenal glands. Eur J Endocrinol 2011; 164: 613-619
    Google Scholar
  • 29 Ye P, Mariniello B, Mantero F, Shibata H, Rainey WE. G-protein-coupled receptors in aldosterone-producing adenomas: a potential cause of hyperaldosteronism. J Endocrinol 2007; 195: 39-48
    CrossrefPubMedGoogle Scholar
  • 30 Parmar J, Key RE, Rainey WE. Development of an adrenocorticotropin-responsive human adrenocortical carcinoma cell line. J Clin Endocrinol Metab 2008; 93: 4542-4546
    CrossrefPubMedGoogle Scholar
  • 31 Kiiveri S, Liu J, Heikkila P, Arola J, Lehtonen E, Voutilainen R, Heikinheimo M. Transcription factors GATA-4 and GATA-6 in human adrenocortical tumors. Endocr Res 2004; 30: 919-923
    CrossrefPubMedGoogle Scholar
  • 32 Schteingart DE, Giordano TJ, Benitez RS, Burdick MD, Starkman MN, Arenberg DA, Strieter RM. Overexpression of CXC chemokines by an adrenocortical carcinoma: a novel clinical syndrome. J Clin Endocrinol Metab 2001; 86: 3968-3974
    CrossrefPubMedGoogle Scholar
  • 33 Almeida MQ, Fragoso MC, Lotfi CF, Santos MG, Nishi MY, Costa MH, Lerario AM, Maciel CC, Mattos GE, Jorge AA, Mendonca BB, Latronico AC. Expression of insulin-like growth factor-II and its receptor in pediatric and adult adrenocortical tumors. J Clin Endocrinol Metab 2008; 93: 3524-3531
    CrossrefPubMedGoogle Scholar
  • 34 Hornsby PJ, Ryan RF, Cheng CY. Replicative senescence and differentiated gene expression in cultured adrenocortical cells. Exp Gerontol 1989; 24: 539-558
    CrossrefPubMedGoogle Scholar
  • 35 Samandari E, Kempna P, Nuoffer JM, Hofer G, Mullis PE, Fluck CE. Human adrenal corticocarcinoma NCI-H295R cells produce more androgens than NCI-H295A cells and differ in 3beta-hydroxysteroid dehydrogenase type 2 and 17,20 lyase activities. J Endocrinol 2007; 195: 459-472
    CrossrefPubMedGoogle Scholar
  • 36 Bird IM, Mason JI, Rainey WE. Regulation of type 1 angiotensin II receptor messenger ribonucleic acid expression in human adrenocortical carcinoma H295 cells. Endocrinology 1994; 134: 2468-2474
    CrossrefPubMedGoogle Scholar
  • 37 Nogueira EF, Vargas CA, Otis M, Gallo-Payet N, Bollag WB, Rainey WE. Angiotensin-II acute regulation of rapid response genes in human, bovine, and rat adrenocortical cells. J Mol Endocrinol 2007; 39: 365-374
    CrossrefPubMedGoogle Scholar
  • 38 Bird IM, Mason JI, Rainey WE. Hormonal regulation of angiotensin II type 1 receptor expression and AT1-R mRNA levels in human adrenocortical cells. Endocr Res 1995; 21: 169-182
    CrossrefPubMedGoogle Scholar
  • 39 Rainey WE, Bird IM, Sawetawan C, Hanley NA, McCarthy JL, McGee EA, Wester R, Mason JI. Regulation of human adrenal carcinoma cell (NCI-H295) production of C19 steroids. J Clin Endocrinol Metab 1993; 77: 731-737
    CrossrefPubMedGoogle Scholar