Aromatase activity induction in human adipose fibroblasts by retinoic acids via retinoic acid receptor α

Wilde, J. de; Erdmann, Maria; Mertens, Michael; Eiselt, Gabriele; Schmidt, Martin

doi:10.1530/jme-12-0129

Cited by 5 publications

(10 citation statements)

References 41 publications

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“…Preparation of soluble nuclear extracts was based on a method published by Wilde et al [19]. The protein concentration was quantified by the Bradford method [20].…”

Section: Preparation Of Soluble Nuclear Extractsmentioning

confidence: 99%

“…Two SNV region-spanning primer sets were used (Supplementary Materials, Table S1). The products were analyzed on 12% polyacrylamide gels stained with ethidium bromide, as described previously [19].…”

Section: Chromatin Immunoprecipitation (Chip)mentioning

confidence: 99%

“…The quantification of full-length aromatase mRNA-expression and utilization of promoters I.3 and II, respectively, was performed by quantitative real-time PCR (qRT-PCR), as described in detail by Wilde et al [19]. Primer sequences are given in the Supplementary Materials, Table S1.…”

Section: Quantification Of Aromatase Mrna-expression In Bafsmentioning

confidence: 99%

“…After 24 h of forskolin stimulation, DNA and mRNA were isolated using the AllPrep DNA/RNA Mini Kit (Qiagen; Hilden, Germany) with on-column DNA-digestion during the RNA-isolation. cDNA was synthesized as described [19]. The ABsolute SYBR Green Rox Mix (Thermo Scientific; Schwerte, Germany) was used for qRT-PCR in a StepOnePlus™ real-time PCR system (Applied Biosystems).…”

Section: Quantification Of Aromatase Promoter-utilization In Transfecmentioning

confidence: 99%

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Identification of PARP-1, Histone H1 and SIRT-1 as New Regulators of Breast Cancer-Related Aromatase Promoter I.3/II

Kaiser

Krüger

Eiselt

et al. 2020

Cells

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Paracrine interactions between malignant estrogen receptor positive (ER+) breast cancer cells and breast adipose fibroblasts (BAFs) stimulate estrogen biosynthesis by aromatase in BAFs. In breast cancer, mainly the cAMP-responsive promoter I.3/II-region mediates excessive aromatase expression. A rare single nucleotide variant (SNV) in this promoter region, which caused 70% reduction in promoter activity, was utilized for the identification of novel regulators of aromatase expression. To this end, normal and mutant promoter activities were measured in luciferase reporter gene assays. DNA-binding proteins were captured by DNA-affinity and identified by mass spectrometry. The DNA binding of proteins was analyzed using electrophoretic mobility shift assays, immunoprecipitation-based in vitro binding assays and by chromatin immunoprecipitation in BAFs in vivo. Protein expression and parylation were analyzed by western blotting. Aromatase activities and RNA-expression were measured in BAFs. Functional consequences of poly (ADP-ribose) polymerase-1 (PARP-1) knock-out, rescue or overexpression, respectively, were analyzed in murine embryonic fibroblasts (MEFs) and the 3T3-L1 cell model. In summary, PARP-1 and histone H1 (H1) were identified as critical regulators of aromatase expression. PARP-1-binding to the SNV-region was crucial for aromatase promoter activation. PARP-1 parylated H1 and competed with H1 for DNA-binding, thereby inhibiting its gene silencing action. In MEFs (PARP-1 knock-out and wild-type) and BAFs, PARP-1-mediated induction of the aromatase promoter showed bi-phasic dose responses in overexpression and inhibitor experiments, respectively. The HDAC-inhibitors butyrate, panobinostat and selisistat enhanced promoter I.3/II-mediated gene expression dependent on PARP-1-activity. Forskolin stimulation of BAFs increased promoter I.3/II-occupancy by PARP-1, whereas SIRT-1 competed with PARP-1 for DNA binding but independently activated the promoter I.3/II. Consistently, the inhibition of both PARP-1 and SIRT-1 increased the NAD+/NADH-ratio in BAFs. This suggests that cellular NAD+/NADH ratios control the complex interactions of PARP-1, H1 and SIRT-1 and regulate the interplay of parylation and acetylation/de-acetylation events with low NAD+/NADH ratios (reverse Warburg effect), promoting PARP-1 activation and estrogen synthesis in BAFs. Therefore, PARP-1 inhibitors could be useful in the treatment of estrogen-dependent breast cancers.

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“…Preparation of soluble nuclear extracts was based on a method published by Wilde et al [19]. The protein concentration was quantified by the Bradford method [20].…”

Section: Preparation Of Soluble Nuclear Extractsmentioning

confidence: 99%

Section: Chromatin Immunoprecipitation (Chip)mentioning

confidence: 99%

Section: Quantification Of Aromatase Mrna-expression In Bafsmentioning

confidence: 99%

Section: Quantification Of Aromatase Promoter-utilization In Transfecmentioning

confidence: 99%

See 2 more Smart Citations

Identification of PARP-1, Histone H1 and SIRT-1 as New Regulators of Breast Cancer-Related Aromatase Promoter I.3/II

Kaiser

Krüger

Eiselt

et al. 2020

Cells

Self Cite

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“…Expression of Cyp19A1 in the adipose tissue of breast cancer subjects is higher at regions proximal to the tumor sites as compared to distal regions. The expression and subsequent activity of Ar is also induced by a higher activation of the retinoic acid receptor-related orphan receptor α (ROR α) in breast cancer cells [113] and by retinoic acid via retinoic acid receptor (RAR)α in human adipocytes [114]. ERβ is also expressed in the human adipose tissue in men and women and its expression has been associated with breast cancer [115].…”

Section: Aromatase Enzyme Complex (Ar) Functions In Adipose Tissuementioning

confidence: 99%

Enzymatic intracrine regulation of white adipose tissue

DiSilvestro

Petrosino

Aldoori

et al. 2014

Hormone Molecular Biology and Clinical Investigation

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Abdominal fat formation has become a permanent risk factor for metabolic syndrome and various cancers in one-third of the world's population of obese and even lean patients. Formation of abdominal fat involves additional mechanisms beyond an imbalance in energy intake and expenditure, which explains systemic obesity. In this review, we briefly summarized autonomous regulatory circuits that locally produce hormones from inactive precursors or nutrients for intra-/auto-/paracrine signaling in white adipose depots. Enzymatic pathways activating steroid and thyroid hormones in adipose depots were compared with enzymatic production of retinoic acid from vitamin A. We discussed the role of intracrine circuits in fat-depot functions and strategies to reduce abdominal adiposity through thermogenic adipocytes with interrupted generation of retinoic acid.

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Human Cytochrome P450 Enzymes

Guengerich

2015

Cytochrome P450

137

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Aromatase activity induction in human adipose fibroblasts by retinoic acids via retinoic acid receptor α

Abstract: Estrogen synthesis in adipose tissue is associated with the development of breast cancer.

Cited by 5 publications

References 41 publications

Identification of PARP-1, Histone H1 and SIRT-1 as New Regulators of Breast Cancer-Related Aromatase Promoter I.3/II

Identification of PARP-1, Histone H1 and SIRT-1 as New Regulators of Breast Cancer-Related Aromatase Promoter I.3/II

Enzymatic intracrine regulation of white adipose tissue

Human Cytochrome P450 Enzymes

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