Adrenocortical cell lines

Rainey, William E.; Saner, Karla; Schimmer, Bernard P.

doi:10.1016/j.mce.2003.12.020

Cited by 207 publications

(182 citation statements)

References 141 publications

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“…Cultured adrenocortical Y-1 cells 17 , which express Ad4BP/SF-1, were used to examine whether cellular proliferation would be affected by knockdown with small interfering RNA (siRNA; mixture of three siRNAs) targeting Ad4BP/SF-1 (siAd4BP/SF-1) or control siRNA (siControl). Treatment with siAd4BP/SF-1 reduced the expression of Ad4BP/SF-1 mRNA and protein by approximately 80% (Fig.…”

Section: Resultsmentioning

confidence: 99%

Glycolytic genes are targets of the nuclear receptor Ad4BP/SF-1

et al. 2014

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Genetic deficiencies in transcription factors can lead to the loss of certain types of cells and tissue. The steroidogenic tissue-specific nuclear receptor Ad4BP/SF-1 (NR5A1) is one such gene, because mice in which this gene is disrupted fail to develop the adrenal gland and gonads. However, the specific role of Ad4BP/SF-1 in these biological events remains unclear. Here we use chromatin immunoprecipitation sequencing to show that nearly all genes in the glycolytic pathway are regulated by Ad4BP/SF-1. Suppression of Ad4BP/SF-1 by small interfering RNA reduces production of the energy carriers ATP and nicotinamide adenine dinucleotide phosphate, as well as lowers expression of genes involved in glucose metabolism. Together, these observations may explain tissue dysgenesis as a result of Ad4BP/SF-1 gene disruption in vivo. Considering the function of estrogen-related receptor a, the present study raises the possibility that certain types of nuclear receptors regulate sets of genes involved in metabolic pathways to generate energy carriers.

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Section: Resultsmentioning

confidence: 99%

Glycolytic genes are targets of the nuclear receptor Ad4BP/SF-1

et al. 2014

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“…The H295R cell line is widely used to study steroid hormone production and to define the molecular mechanisms regulating transcription of steroidogenic enzymes (Rainey et al 2004. Although the H295R cells increase aldosterone production in response to Ang II and potassium treatment , Clyne et al 1997, the cells have only moderate responsiveness to ACTH.…”

Section: Disscusionmentioning

confidence: 99%

“…In vitro cell culture models provide important information for understanding molecular and cellular mechanisms controlling both the normal and pathologic function of the adrenal cortex. The H295R adrenocortical cell line is widely used but is known to have only modest ACTH responsiveness , Rainey et al 2004. Therefore, studies directed toward ACTH action have used primary cultures of adrenal cells (Xing et al 2010(Xing et al , 2011 or Y-1 mouse adrenal cell line, which retains ACTH responsiveness (Schimmer 1979).…”

mentioning

confidence: 99%

H295R expression of melanocortin 2 receptor accessory protein results in ACTH responsiveness

Nanba¹,

Chen²,

Turcu³

et al. 2015

Journal of Molecular Endocrinology

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The H295R adrenocortical cell line is widely used for molecular analysis of adrenal functions but is known to have only modest ACTH responsiveness. The lack of ACTH response was linked to a low expression of its receptor, melanocortin 2 receptor (MC2R). We hypothesized that increasing the MC2R accessory protein (MRAP), which is required to traffic MC2R from the endoplasmic reticulum to the cell surface, would increase ACTH responsiveness. Lentiviral particles containing human MRAP-open reading frame were generated and transduced in H295R cells. Using antibiotic resistance, 18 clones were isolated for characterization. The most ACTH-responsive steroidogenic clone, H295RA, was used for further experiments. Successful induction of MRAP and increased expression of MC2R in H295RA cells was confirmed by quantitative real-time RT-PCR and protein analysis. Treatment with ACTH significantly increased aldosterone, cortisol, and dehydroepiandrosterone production in H295RA cells. ACTH also significantly increased transcript levels for all of the steroidogenic enzymes required to produce aldosterone, cortisol, and dehydroepiandrosterone, as well as MC2R mRNA. Using liquid chromatography/tandem mass spectrometry, we further revealed that the main unconjugated steroids produced in H295RA cells were 11-deoxycortisol, cortisol, and androstenedione. Treatment of H295RA cells with ACTH also acutely increased cAMP production and cellular protein levels for total and phosphorylated steroidogenic acute regulatory protein. In summary, through genetic manipulation, we have developed an ACTH-responsive human adrenocortical cell line. The cell line will provide a powerful in vitro tool for molecular analysis of physiologic and pathologic conditions involving the hypothalamic-pituitary-adrenal axis.

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“…To this end, we used a human adrenocarcinoma (H295R) cell line, 23 and the depolarizing response of N-type calcium channels was confirmed by patch clamp techniques. In addition, the steroidogenesis response of N-type calcium channels was examined by using omega-conotoxin GVIA (CnTX; an N-type calcium channel inhibitor) and other CCBs such as cilnidipine (L-type/N-type CCB), nifedipine (L-type CCB), and efonidipine (L-type/T-type CCB).…”

Section: Introductionmentioning

confidence: 99%

Expression of N-type calcium channels in human adrenocortical cells and their contribution to corticosteroid synthesis

et al. 2010

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The inhibition of aldosterone activity is a useful approach for preventing the progression of cardiovascular and renal diseases in hypertensive patients. Although the results of our previous in vivo study suggested that N-type calcium channels may have a role in regulating plasma aldosterone levels, the direct relationship between N-type calcium channels and aldosterone production in adrenocortical cells has not been examined. In this study, the analysis of quantitative reverse transcription-PCR, western blotting, and immunocytological staining indicated the possible presence of N-type calcium channels in human adrenocortical cells (H295R cell line). Patch clamp analysis indicated that omega-conotoxin GVIA (CnTX), an N-type calcium channel inhibitor, suppressed voltage-dependent barium currents. During steroidogenesis, CnTX significantly reduced the transient calcium signaling induced by angiotensin II (Ang II) and partially prevented Ang II-induced aldosterone and cortisol formation with no significant influence on CYP11B2 and CYP11B1 mRNA expression. In addition, in a1B calcium channel subunits, knockdown significantly decreased Ang II-induced aldosterone formation with increments in CYP11B2 mRNA expression. We also investigated the inhibitory activities of some types of dihydropyridine calcium channel blockers (CCBs; cilnidipine: L-/N-type CCB, efonidipine: L-/T-type CCB, and nifedipine: L-type CCB), and these agents showed a dose-dependent inhibition effect on Ang II-induced aldosterone and cortisol production. Furthermore, only cilnidipine failed to suppress CYP11B1 expression in H295R cells. These results suggest that N-type calcium channels have a significant role in transducing the Ang II signal for aldosterone (and cortisol) biosynthesis, which may explain the mechanism by which N-type calcium channels regulate plasma aldosterone levels.

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Adrenocortical cell lines

Cited by 207 publications

References 141 publications

Glycolytic genes are targets of the nuclear receptor Ad4BP/SF-1

Glycolytic genes are targets of the nuclear receptor Ad4BP/SF-1

H295R expression of melanocortin 2 receptor accessory protein results in ACTH responsiveness

Expression of N-type calcium channels in human adrenocortical cells and their contribution to corticosteroid synthesis

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