Expression of Steroidogenic Factor 1 (SF-1) and Steroidogenic Acute Regulatory Protein (StAR) in Endometriosis is Associated with Endometriosis Severity

Basal plasma ACTH1–39 concentrations are elevated in long-term hypoxic (LTH) fetal sheep. This study was designed to determine whether the expression of genes regulating cortisol biosynthesis was altered after LTH. Pregnant ewes were maintained at high altitude (3,820 m) from day 30 of gestation to near term, when the animals were transported to the laboratory. Reduced Po2 was maintained by nitrogen infusion through a maternal tracheal catheter. On days 137–141, fetal adrenal glands were collected from LTH and normoxic control fetuses. Real-time PCR was used to quantify mRNA for steroidogenic acute regulatory protein, 17α-hydroxylase (CYP17), 21-hydroxylase (CYP21), cholesterol side-chain cleavage (CYP11A1), 3β-hydroxysteroid dehydrogenase type II (HSD3B2), and the ACTH receptor. We analyzed mRNA by slot-blot hybridization and also quantified mRNA for transcription factors necessary for adrenocortical development by quantitative real-time PCR: steroidogenic factor 1 and dosage-sensitive sex reversal, adrenal hypoplasia congenital, critical region on the X chromosome (DAX-1). Protein was quantified by Western blot analysis. Adrenal mRNAs for CYP17, CYP11A1, and the ACTH receptor were significantly reduced in LTH fetal sheep compared with levels shown in controls. Similarly, CYP11A1 protein and CYP17 protein were reduced in the LTH group. CYP21, steroidogenic acute regulatory protein, HSD3B2, steroidogenic factor 1, and DAX-1 expressions were not altered in response to LTH. We conclude that expression of two key steroidogenic enzymes (CYP17, CYP11A1) regulating cortisol biosynthesis and the ACTH receptor is lower in response to LTH. This likely represents an adaptive response to LTH, to prevent excessive cortisol production that would restrict fetal growth and potentially induce preterm delivery.

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Steroidogenic Factor 1-Dependent Promoter Activity of the Human Steroidogenic Acute Regulatory Protein (StAR) Gene†

Biochemistry ◽

10.1021/bi960057r ◽

1996 ◽

Vol 35 (28) ◽

pp. 9052-9059 ◽

Cited By ~ 182

Author(s):

Teruo Sugawara ◽

John A. Holt ◽

Marianthi Kiriakidou ◽

Jerome F. Strauss

Keyword(s):

Promoter Activity ◽

Regulatory Protein ◽

Steroidogenic Acute Regulatory Protein ◽

Steroidogenic Factor 1 ◽

Steroidogenic Acute Regulatory ◽

Star Gene

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Screening for Mutations in the Steroidogenic Acute Regulatory Protein and Steroidogenic Factor-1 Genes, and in CYP11A and Dosage-Sensitive Sex Reversal-Adrenal Hypoplasia Gene on the X Chromosome, Gene-1 (DAX-1), in Hyperandrogenic Hirsute Women1

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.86.4.7424 ◽

2001 ◽

Vol 86 (4) ◽

pp. 1746-1749

Author(s):

Rosa M. Calvo ◽

Miryam Asunción ◽

Dolores Tellería ◽

José Sancho ◽

José L. San Millán ◽

...

Keyword(s):

X Chromosome ◽

Regulatory Protein ◽

Sex Reversal ◽

Steroidogenic Acute Regulatory Protein ◽

Steroidogenic Factor 1 ◽

Chromosome Gene ◽

Adrenal Hypoplasia ◽

Steroidogenic Acute Regulatory

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Screening for Mutations in the Steroidogenic Acute Regulatory Protein and Steroidogenic Factor-1 Genes, and in CYP11A and Dosage-Sensitive Sex Reversal-Adrenal Hypoplasia Gene on the X Chromosome, Gene-1 (DAX-1), in Hyperandrogenic Hirsute Women

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.86.4.1746 ◽

2001 ◽

Vol 86 (4) ◽

pp. 1746-1749 ◽

Cited By ~ 10

Author(s):

R. M. Calvo

Keyword(s):

X Chromosome ◽

Regulatory Protein ◽

Sex Reversal ◽

Steroidogenic Acute Regulatory Protein ◽

Steroidogenic Factor 1 ◽

Chromosome Gene ◽

Adrenal Hypoplasia ◽

Steroidogenic Acute Regulatory

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Multiple Steroidogenic Factor 1 Binding Elements in the Human Steroidogenic Acute Regulatory Protein Gene 5‘-Flanking Region Are Required for Maximal Promoter Activity and Cyclic AMP Responsiveness†

Biochemistry ◽

10.1021/bi9628984 ◽

1997 ◽

Vol 36 (23) ◽

pp. 7249-7255 ◽

Cited By ~ 120

Author(s):

Teruo Sugawara ◽

Marianthi Kiriakidou ◽

Jan M. McAllister ◽

Caleb B. Kallen ◽

Jerome F. Strauss

Keyword(s):

Cyclic Amp ◽

Promoter Activity ◽

Protein Gene ◽

Regulatory Protein ◽

Steroidogenic Acute Regulatory Protein ◽

Steroidogenic Factor 1 ◽

Flanking Region ◽

Steroidogenic Acute Regulatory

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Differentiation of Human Embryonic Stem Cells and Human Induced Pluripotent Stem Cells into Steroid-Producing Cells

Endocrinology ◽

10.1210/en.2012-1060 ◽

2012 ◽

Vol 153 (9) ◽

pp. 4336-4345 ◽

Cited By ~ 33

Author(s):

Takuhiro Sonoyama ◽

Masakatsu Sone ◽

Kyoko Honda ◽

Daisuke Taura ◽

Katsutoshi Kojima ◽

...

Keyword(s):

Stem Cells ◽

Regulatory Protein ◽

Embryonic Stem ◽

Ips Cells ◽

Steroidogenic Acute Regulatory Protein ◽

Steroidogenic Factor 1 ◽

Differentiated Cells ◽

Steroidogenic Acute Regulatory ◽

Induced Pluripotent ◽

Mesodermal Lineage

Although there have been reports of the differentiation of mesenchymal stem cells and mouse embryonic stem (ES) cells into steroid-producing cells, the differentiation of human ES/induced pluripotent stem (iPS) cells into steroid-producing cells has not been reported. The purpose of our present study was to establish a method for inducing differentiation of human ES/iPS cells into steroid-producing cells. The first approach we tried was embryoid body formation and further culture on adherent plates. The resultant differentiated cells expressed mRNA encoding the steroidogenic enzymes steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, cytochrome P450-containing enzyme (CYP)-11A1, CYP17A1, and CYP19, and secreted progesterone was detected in the cell medium. However, expression of human chorionic gonadotropin was also detected, suggesting the differentiated cells were trophoblast like. We next tried a multistep approach. As a first step, human ES/iPS cells were induced to differentiate into the mesodermal lineage. After 7 d of differentiation induced by 6-bromoindirubin-3′-oxime (a glycogen synthase kinase-3β inhibitor), the human ES/iPS cells had differentiated into fetal liver kinase-1- and platelet derived growth factor receptor-α-expressing mesodermal lineage cells. As a second step, plasmid DNA encoding steroidogenic factor-1, a master regulator of steroidogenesis, was introduced into these mesodermal cells. The forced expression of steroidogenic factor-1 and subsequent addition of 8-bromoadenosine 3′,5′-cyclic monophosphate induced the mesodermal cells to differentiate into the steroidogenic cell lineage, and expression of CYP21A2 and CYP11B1, in addition to steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, CYP11A1, and CYP17A1, was detected. Moreover, secreted cortisol was detected in the medium, but human chorionic gonadotropin was not. These findings indicate that the steroid-producing cells obtained through the described multistep method are not trophoblast like; instead, they exhibit characteristics of adrenal cortical cells.

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Salt-Inducible Kinase Is Involved in the ACTH/cAMP-Dependent Protein Kinase Signaling in Y1 Mouse Adrenocortical Tumor Cells

Molecular Endocrinology ◽

10.1210/mend.15.8.0675 ◽

2001 ◽

Vol 15 (8) ◽

pp. 1264-1276 ◽

Cited By ~ 32

Author(s):

Xing-zi Lin ◽

Hiroshi Takemori ◽

Yoshiko Katoh ◽

Junko Doi ◽

Nanao Horike ◽

...

Keyword(s):

Cytochrome P450 ◽

Promoter Activity ◽

Regulatory Protein ◽

Steroidogenic Acute Regulatory Protein ◽

Side Chain ◽

Mrna Levels ◽

Steroidogenic Factor 1 ◽

Side Chain Cleavage ◽

Chain Cleavage ◽

Steroidogenic Acute Regulatory

Abstract The involvement of salt-inducible kinase, a recently cloned protein serine/threonine kinase, in adrenal steroidogenesis was investigated. When Y1 mouse adrenocortical tumor cells were stimulated by ACTH, the cellular content of salt-inducible kinase mRNA, protein, and enzyme activity changed rapidly. Its level reached the highest point in 1–2 h and returned to the initial level after 8 h. The mRNA levels of cholesterol side-chain cleavage cytochrome P450 and steroidogenic acute regulatory protein, on the other hand, began to rise after a few hours, reaching the highest levels after 8 h. The salt-inducible kinase mRNA level in ACTH-, forskolin-, or 8-bromo-cAMP-treated Kin-7 cells, mutant Y1 with less cAMP-dependent PKA activity, remained low. However, Kin-7 cells, when transfected with a PKA expression vector, expressed salt-inducible kinase mRNA. Y1 cells that overexpressed salt-inducible kinase were isolated, and the mRNA levels of steroidogenic genes in these cells were compared with those in the parent Y1. The level of cholesterol side-chain cleavage cytochrome P450 mRNA in the salt-inducible kinase-overexpressing cells was markedly low compared with that in the parent, while the levels of Ad4BP/steroidogenic factor-1-, ACTH receptor-, and steroidogenic acute regulatory protein-mRNAs in the former were similar to those in the latter. The ACTH-dependent expression of cholesterol side-chain cleavage cytochrome P450- and steroidogenic acute regulatory protein-mRNAs in the salt-inducible kinase-overexpressing cells was significantly repressed. The promoter activity of the cholesterol side-chain cleavage cytochrome P450 gene was assayed by using Y1 cells transfected with a human cholesterol side-chain cleavage cytochrome P450 promoter-linked reporter gene. Addition of forskolin to the culture medium enhanced the cholesterol side-chain cleavage cytochrome P450 promoter activity, but the forskolin-dependently activated promoter activity was inhibited when the cells were transfected with a salt-inducible kinase expression vector. This inhibition did not occur when the cells were transfected with a salt-inducible kinase (K56M) vector that encoded an inactive kinase. The salt-inducible kinase’s inhibitory effect was also observed when nonsteroidogenic, nonAd4BP/steroidogenic factor-1 -expressing, NIH3T3 cells were used for the promoter assays. These results suggested that salt-inducible kinase might play an important role(s) in the cAMP-dependent, but Ad4BP/steroidogenic factor-1-independent, gene expression of cholesterol side-chain cleavage cytochrome P450 in adrenocortical cells.

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