Cited by 52Hormonal Regulation and HypertensionRenin-Angiotensin System StudiesAdrenal Hormones and Disorders

Differential effects of high and low steroidogenic factor-1 expression on CYP11B2 expression and aldosterone production in adrenocortical cells

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Ye, Ping, Nakamura, Yashuhiro, Lalli, Enzo, Rainey, William E.·2009·Endocrinology

Abstract

Steroidogenic factor-1 (SF-1/Ad4BP/NR5A1) plays a major role in regulating steroidogenic enzymes. We have previously shown that SF-1 inhibits aldosterone synthase (CYP11B2) reporter gene activity. Herein, we used the H295R/TR/SF-1 adrenal cells that increase SF-1 in a doxycycline-dependent fashion. Cells were incubated with or without doxycycline to induce SF-1 and then treated with angiotensin II (Ang II). Aldosterone was measured by immunoassay. SF-1 mRNA was silenced by small interfering RNA (siRNA) by Nucleofector technology. mRNA levels were measured by real-time RT-PCR. Ang II treatment without doxycycline increased aldosterone production by 11.3-fold and CYP11B2 mRNA by 116-fold. Doxycycline treatment increased SF-1 mRNA levels by 3.7-fold and inhibited Ang II-induced aldosterone by 84%. Doxycycline treatment inhibited Ang II-stimulated CYP11B2 mRNA levels by 86%. Doxycycline decreased basal CYP11B2 promoter activity by 68%. Doxycycline inhibited Ang II stimulation by 85%. Ang II increased CYP21 mRNA expression by 4.6-fold, whereas doxycycline inhibited induction by 69%. In contrast, doxycycline treatment increased CYP11B1 mRNA by 1.7-fold in basal cells and increased Ang II induction by 3.6-fold. SF-1-specific siRNA significantly reduced SF-1 mRNA expression as compared with cells treated with control siRNA. SF-1 siRNA reversed doxycycline stimulation of CYP B1 and its inhibition of CYP11B2. However, in H295R/TR/SF-1 cells without doxycycline treatment, both CYP11B1 and CYP11B2 mRNAs were significantly decreased, suggesting that both enzymes require a minimal level of SF-1 for basal expression. In summary, SF-1 overexpression dramatically inhibited CYP11B2 expression and decreased aldosterone production. The opposing effects of SF-1 on CYP11B1 and CYP11B2 suggest that the regulation of SF-1 activity may play a role that determines the relative ability to produce mineralocorticoid and glucocorticoid.

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Crossref · 10.1210/en.2008-0667

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Authors
Ye, P., Nakamura, Y., Lalli, E., & Rainey, W. E.
Affiliations
  • Augusta University (US)
  • Centre National de la Recherche Scientifique (FR)
Corresponding author
Rainey, William E.
Venue
Endocrinology
Published
2008-10-30
Volume
150
Issue
3
Pages
1303-1309
ISSN
  • 0013-7227 (ISSN-L)
  • 1945-7170
OpenAlex
W2129925307
Cited by
52
References
49
Citations by year
2025: 2 · 2023: 1 · 2022: 3 · 2021: 1 · 2020: 1 · 2019: 1 · 2018: 2 · 2017: 1 · 2016: 5 · 2015: 4 · 2014: 9 · 2013: 5 · 2012: 5
FWCI
2.90
Type
journal article
Language
en
Open access
Locations
  • Endocrinology (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • HAL (submittedVersion)
Funders
  • National Institutes of Health
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK069950, DK 43140, R01DK069950, R01DK043140)
Topics
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal Regulation and Hypertension
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Renin-Angiotensin System Studies
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
Keywords
steroidogenic factor 1, aldosterone synthase, CYP11B2, CYP11B1, CYP21, aldosterone biosynthesis, angiotensin II, adrenocortical cells, H295R cells, doxycycline, siRNA knockdown, steroidogenesis regulation
MeSH terms
Adrenal Cortex, Aldosterone, Angiotensin II, Cells, Cultured, Doxycycline, Gene Expression Regulation, Glucocorticoids, Humans, Mineralocorticoids, Steroid 11-beta-Hydroxylase, Cytochrome P-450 CYP11B2, RNA, Small Interfering, Steroidogenic Factor 1

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