DOCSLIB.ORG

1 ,25-Dihydroxyvitamin D3 Down-Regulates Estrogen Receptor Abundance and Suppresses Estrogen Actions in MCF-7 Human Breast Cancer Cells1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
1 ,25-Dihydroxyvitamin D3 Down-Regulates Estrogen Receptor Abundance and Suppresses Estrogen Actions in MCF-7 Human Breast Cancer Cells1 Vol. 6, 3371–3379, August 2000 Clinical Cancer Research 3371 ␣ 1 ,25-Dihydroxyvitamin D3 Down-Regulates Estrogen Receptor Abundance and Suppresses Estrogen Actions in MCF-7 Human Breast Cancer Cells1 Srilatha Swami, Aruna V. Krishnan, and increase in breast cancer susceptibility gene (BRCA1) pro- 2 David Feldman tein is reduced by 1,25(OH)2D3 treatment. Overall, these Department of Medicine, Stanford University School of Medicine, results suggest that the antiproliferative effects of Stanford, California 94305 1,25(OH)2D3 and its analogues on MCF-7 cells could par- tially be mediated through their action to down-regulate ER levels and thereby attenuate estrogenic bioresponses, includ- ABSTRACT ing breast cancer cell growth. ␣ 1 ,25-Dihydroxyvitamin D3 [1,25(OH)2D3], the active metabolite of vitamin D, is a potent inhibitor of breast cancer cell growth. Because the estrogen receptor (ER) plays INTRODUCTION a key role in breast cancer progression, we have studied the Breast cancer is the most commonly diagnosed cancer and effects of 1,25(OH)2D3 on the regulation of ER in the estro- the second leading cause of cancer-related deaths among women gen-responsive MCF-7 human breast cancer cell line, which in the United States (1). Because breast cancer is generally ␣ is known to predominantly express ER . 1,25(OH)2D3 characterized by estrogen-dependent growth, the abundance of 3 causes significant inhibition of MCF-7 cell growth, and it ERs in these cells assumes critical importance (2). E2 acts via also decreases the growth-stimulatory effect of 17␤-estradiol the ER, a member of the steroid/thyroid/retinoid receptor super- (E2). Treatment of MCF-7 cells with 1,25(OH)2D3 reduces family (3). The factors and mechanisms that control the level of ER levels in a dose-dependent manner, as shown by ligand ER expression are important in determining the amplitude of binding assays and Western blot analysis. The 1,25(OH)2D3 E2-mediated actions on the breast cancer cells (2). analogues EB-1089, KH-1060, Ro 27-0574, and Ro 23-7553 1,25(OH)2D3, the biologically active form of vitamin D, is are more potent than 1,25(OH)2D3 in both their antiprolif- a major regulator of calcium and phosphate homeostasis in the erative actions as well as ER down-regulation. There is a body (4, 5). The regulatory effects of 1,25(OH)2D3 are mediated /between the growth-inhibi- via the VDR, which is also a member of the steroid/thyroid (0.98 ؍ striking correlation (R2 tory actions of 1,25(OH)2D3 or analogues and their ability to retinoid receptor superfamily (4–6). In addition to its effects on down-regulate ER levels. Treatment with 1,25(OH)2D3 calcium and phosphate homeostasis, 1,25(OH)2D3 is an impor- shows that the reduction in ER is accompanied by a signif- tant modulator of cellular proliferation and differentiation in a icant decrease in the steady-state levels of ER mRNA. The number of normal and malignant cells (4, 7–9). In breast cancer decrease in ER mRNA is not abolished by the protein syn- cells, 1,25(OH)2D3 has potent growth-inhibitory actions (10– thesis inhibitor cycloheximide. Inhibition of mRNA synthe- 13). Although the growth-inhibitory effects of 1,25(OH)2D3 on sis with actinomycin D reveals no significant differences breast cancer cells have been well established, the effects of between ER mRNA half-life in control and 1,25(OH)2D3- 1,25(OH)2D3 on ER expression are less well documented. Stud- treated cells. Nuclear run-on experiments demonstrate sig- ies on ER␣ in human breast cancer cell lines have reported nificant decreases in ER gene transcription at the end of 17 h minor decreases (14) or no change (15) in ER expression with of treatment with 1,25(OH)2D3. These findings indicate that 1,25(OH)2D3 treatment. A more recent study reported signifi- 1,25(OH)2D3 exerts a direct negative effect on ER gene cant decreases in ER protein levels in the MCF-7 cells treated transcription. Coincident with the decrease in ER levels with EB-1089, a potent analogue of 1,25(OH)2D3 (13). Studies there is an attenuation of E2-mediated bioresponses after have also been conducted to indicate that the E2-mediated 1,25(OH)2D3 treatment. Induction of progesterone receptor bioresponses are attenuated by 1,25(OH)2D3 treatment (16, 17). by E2 is suppressed by 1,25(OH)2D3, and the E2-mediated Although the above-mentioned reports have suggested potential cross-talk between 1,25(OH)2D3 and estrogen signaling path- ways, the extent of the interaction and the mechanism by which 1,25(OH)2D3 causes down-regulation of ER are still not clarified. Received 2/14/00; revised 5/5/00; accepted 5/11/00. The purpose of the present investigation is to study the The costs of publication of this article were defrayed in part by the effects of 1,25(OH)2D3 on ER and E2-mediated effects in payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 Supported by NIH Grant DK42482 and Department of the Army Grant DAMD 17-98-8556. 2 3 ␤ To whom requests for reprints should be addressed, at Division of The abbreviations used are: ER, estrogen receptor; E2,17 -estradiol; ␣ Endocrinology, SUMC, Room S-005, Stanford, CA 94305-5103. Phone: 1,25(OH)2D3,1 ,25-dihydroxyvitamin D3; VDR, vitamin D receptor; (650) 725-2910; Fax: (650) 725-7085. E-mail: feldman@cmgm. nVDRE, negative vitamin D response element; ERE, estrogen response stanford.edu. element; CSS, charcoal-stripped serum; PR, progesterone receptor. Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2000 American Association for Cancer Research. 3372 1,25(OH)2D3 Regulates ER in MCF-7 Cells MCF-7 breast cancer cells. As discussed below, the MCF-7 cells Lincoln Park, NJ) at a density of 50,000 cells/well in 3 ml of used in this study do not express ER␤, as measured by reverse RPMI 1640 containing 10% calf serum. Twenty-four h later, transcription-PCR. Therefore, in these studies the effect is lim- fresh medium was added. Cells were grown in RPMI 1640 ited to ER␣. For simplicity, we refer to the ER␣ in these cells as medium with 10% calf serum or CSS and were treated with ER. To achieve our goal of investigating the effect of various doses of 1,25(OH)2D3 or its analogues in the presence or M 1,25(OH)2D3 on breast cancer cells, we have studied the rela- absence of 10 n E2. Fresh medium and hormones were added tionship between changes in MCF-7 growth rate, levels of ER every other day. At the end of 6 days, cell monolayers were protein, steady-state ER mRNA, and gene transcription in cells processed as described earlier (19), and DNA contents were determined by the method of Burton (20). treated with 1,25(OH)2D3 or its analogues. We have also as- Ligand Binding Assays. MCF-7 cells growing in CSS- sessed the correlation between the effects of 1,25(OH)2D3 and its analogues KH-1060, EB-1089, Ro 27-0574, and Ro 23-7553 containing medium were treated with either E2 (10 nM)or on the growth of MCF-7 cells and the changes elicited in ER 1,25(OH)2D3 or analogues (1, 10, or 100 nM) for 2 days. Cells levels. Furthermore, we have investigated how changes in ER were then harvested, and high salt cell extracts were made as described previously (21). The protein concentration of the abundance induced by 1,25(OH)2D3 and its analogues alter the extracts was measured by the method of Bradford (22). Aliquots functional responses to E2 in MCF-7 cells. We have established of the extracts were incubated overnight at 4°C with either 10 that the 1,25(OH)2D3-mediated effect on ER gene expression is 3 3 at the transcriptional level. nM [ H]E2 or 10 nM [ H]progesterone for ER and PR measure- ments, respectively. Two hundred-fold excess of nonradioactive hormone was used to correct for nonspecific binding. Bound and MATERIALS AND METHODS free hormones were separated using hydroxylapatite, and spe- 3 Materials. [ H]Estradiol-17␤-D-glucuronide (specific cific binding was calculated as described earlier (21). activity, 40 Ci/mmol) and [3H]progesterone (specific activity, Western Blot Analysis. Aliquots of cell extracts pre- 54.1 Ci/mmol) were purchased from DuPont NEN (Wilmington, pared as described above were mixed with 3ϫ SDS sample DE). Radioinert steroids were obtained from Steraloids, Inc. buffer, boiled for 5 min, and subjected to 10% SDS-PAGE. (Wilton, NH). Nonradioactive 1,25(OH)2D3 and its analogues After transfer to nitrocellulose membranes, immunoblotting 1,25-dihydroxy-16-ene-23-yne-cholecalciferol (Ro 23-7553) with either the antimouse monoclonal antibody to human ER and 1,25-dihydroxy-23-yne-26,27-hexafluoro-20-cyclopropyl- (H222; 1:500 dilution in 1% Carnation nonfat milk; a gift from 19-nor-cholecalciferol (Ro 27-0574) were generous gifts Abbott laboratories) or antimouse monoclonal antibody to the from Dr. M. Uskokovic (Hoffmann La-Roche Co., Nutley, human BRCA1 protein (C-20; 2 ␮g/ml in 1% Carnation nonfat NJ). 1␣,25-Dihydroxy-22,24-diene-24,26,27-trihomovitamin milk; from Santa Cruz Biotechnology) was carried out as de- ␣ D3 (EB-1089) and 1 ,25-dihydroxy-20-epi-22-ene-24,26,27- scribed previously (19).
Load more

Recommended publications
  • What Can I Do to Protect Myself and My Family During COVID-19?
    What Can I Do to Protect Role for Myself, My Complementary Family and My and Integrative Patients During Health? COVID-19? Karen H. Seal, MD, MPH Chief, Integrative Health Service San Francisco VA Medical Center Professor of Medicine and Psychiatry, UCSF Disclaimer 1. Material presented here is supplemental; does not replace public health guidelines emphasizing hand-washing, masking, social distancing and testing if symptoms present. 2. No CIH measures have been validated in human trials as effective against COVID-19; evidence is from pre-clinical trials or trials related to boosting immunity in general. 3. Opinions expressed here do not reflect those of the Department of Veterans Affairs or the University of California, San Francisco. Poll Question #1 In addition to public health measures (masking, distancing, handwashing etc.), have you implemented anything new in an attempt to protect you or family members against COVID-19? YES NO Poll Question # 2 If you have implemented new health measures, in which of the following areas have you made changes? (Indicate all that apply.) A. Sleep habits B. Diet or dietary supplements (vitamins, minerals, other natural products) C. Exercise D. Stress Reduction (other than exercise) E. None of the above Overview – Virology and pathogenicity of SARS-CoV-2 (Coronavirus disease 2019; COVID-19) – Strategies to help prevent infection and promote recovery – Nutrition – Dietary Supplements – Sleep – Exercise and Stress Reduction – Summary Virology and Pathogenicity of COVID-19 SARS-CoV-2 RNA virus enters human cells through angiotensin-converting–enzyme 2 (ACE2) receptors SARS-CoV-2 virus penetrates host cell Main Goal of Virus injects its blueprint Viruses: Hijacks cell’s “factory” to make new virus proteins Reproduction Assembles new SARS-CoV-2 virions Mature RNA virus released into bloodstream infects other cells Virology of COVID-19 – Virulence and pathogenicity develops from COVID-19 virus activating cytoplasmic NLRP3 inflammasome within immune cells.
    [Show full text]
  • Association of Hypogonadism with Vitamin D Status
    European Journal of Endocrinology (2012) 166 77–85 ISSN 0804-4643 CLINICAL STUDY Association of hypogonadism with vitamin D status: the European Male Ageing Study David M Lee, Abdelouahid Tajar, Stephen R Pye, Steven Boonen1, Dirk Vanderschueren2, Roger Bouillon3, Terence W O’Neill, Gyorgy Bartfai4, Felipe F Casanueva5,6, Joseph D Finn7, Gianni Forti8, Aleksander Giwercman9, Thang S Han10, Ilpo T Huhtaniemi11, Krzysztof Kula12, Michael E J Lean13, Neil Pendleton14, Margus Punab15 and Frederick C W Wu7, the EMAS study group† Arthritis Research UK Epidemiology Unit, Manchester Academic Health Science Centre, The University of Manchester, Manchester M13 9PT, UK, 1Division of Gerontology and Geriatrics and Centre for Musculoskeletal Research, Department of Experimental Medicine, 2Department of Andrology and Endocrinology and 3Department of Experimental Medicine, Katholieke Universiteit Leuven, Leuven B-3000, Belgium, 4Department of Obstetrics, Gynaecology and Andrology, Albert Szent-Gyorgy Medical University, Szeged H-6725, Hungary, 5Department of Medicine, Santiago de Compostela University, Complejo Hospitalario Universitario de Santiago (CHUS), 15076 Santiago de Compostela, Spain, 6CIBER de Fisiopatologı´a Obesidad y Nutricion (CB06/03), Instituto Salud Carlos III, 15076 Santiago de Compostela, Spain, 7Developmental and Regenerative Biomedicine Research Group, Andrology Research Unit, Manchester Academic Health Science Centre, Manchester Royal Infirmary, The University of Manchester, Grafton Street, Manchester M13 9WL, UK, 8Andrology Unit,
    [Show full text]
  • A Novel Androgen–Vitamin D Link
    RESEARCH HIGHLIGHTS Nature Reviews Endocrinology | Published online 09 Feb 2018; doi:10.1038/nrendo.2018.14 BONE A novel androgen–vitamin D link Androgen deficiency in men, which observations were corroborated in mice, suggesting that androgen can result from normal ageing or human cell lines. stimulation decreases 24-hydroxy- prostate cancer therapy, is associated Next, the effect of PR loss on lase-dependent vitamin D3 with reduced levels of vitamin D3 and vitamin D-related gene expression inactivation. Indeed, DHT increased an increased risk of osteoporosis. Now, was investigated. In mouse proximal serum levels of vitamin D3 in these new research has uncovered a novel convoluted tubule (PCT) cells, mice, confirming the link between link between vitamin D3 metabolism Pgr-targeted small interfering RNA androgens and vitamin D homeostasis. and sex hormone levels, a finding that (siRNA) reduced levels of Cyp24a1 “Using modulators of PR, we want could be exploited therapeutically to (encoding 24-hydroxylase), a vitamin to further investigate the role of PR in treat vitamin D deficiency. D3-inactivating enzyme, whereas the metabolism of vitamin D,” explains Eui-Ju Hong and colleagues first human PGR overexpression increased Hong. “We also wish to investigate the assessed whether endogenous Cyp24a1 expression. Subsequent biosynthesis and metabolism of androgens altered androgen- ChIP-sequencing showed that vitamin D in renal tissue during responsive gene expression in the progesterone treatment increased PR pregnancy, during which the kidney, a known androgen target recruitment to the Cyp24a1 expression of sex steroid hormones organ. Dihydrotestosterone (DHT) promoter, inferring a transcriptional and their receptors is increased.” treatment markedly decreased renal mechanism for DHT-dependent Conor A.
    [Show full text]
  • Activin and Estrogen Crosstalk Regulates Transcription in Human Breast Cancer Cells
    Endocrine-Related Cancer (2007) 14 679–689 Activin and estrogen crosstalk regulates transcription in human breast cancer cells Joanna E Burdette1,2 and Teresa K Woodruff1,2,3 1Feinberg College of Medicine, Institute for Women’s Health Research, Northwestern University, Chicago, Illinois 60611, USA 2Center for Reproductive Science, Northwestern University, Evanston, Illinois 60208, USA 3Robert H Lurie Comprehensive Cancer Center of Northwestern University, O T Hogan 4-150, Chicago, Illinois 60611, USA (Correspondence should be addressed to T K Woodruff; Email: [email protected]) Abstract Activin is a member of the transforming growth factor b superfamily that regulates mammary cell function during development, lactation, and in cancer. Activin slows the growth of breast cancer cells by inducing G0/G1 cell cycle arrest. Estrogen is a steroid hormone that stimulates the proliferation of mammary epithelial cells in development and oncogenesis. The crosstalk between estrogen and activin that regulates activin ligand expression, activin and estrogen signal transduction, and cell cycle arrest was investigated in this study. Estrogen antagonized activin- dependent production of plasminogen activator inhibitor 1 (PAI-1) mRNA, while activin repressed estrogen-dependent transcription of trefoil factor 1. The repression of estrogen signaling by activin was recapitulated using a simple estrogen response element-luciferase construct and was enhanced in the presence of overexpressed estrogen receptor a (ERa). In contrast, estrogen- mediated repression of activin signaling could not be recapitulated on a simple CAGA Smad- binding element but did inhibit the short PAI-1 promoter, p3TP-luciferase, especially when ERa was overexpressed. Repression of both estrogen- and activin-regulated transcription was found to be ligand induced and Smad3 dependent.
    [Show full text]
  • Isolation and Characterisation of Lymphatic Endothelial Cells From
    www.nature.com/scientificreports OPEN Isolation and characterisation of lymphatic endothelial cells from lung tissues afected by lymphangioleiomyomatosis Koichi Nishino1,2*, Yasuhiro Yoshimatsu3,4, Tomoki Muramatsu5, Yasuhito Sekimoto1,2, Keiko Mitani1,2, Etsuko Kobayashi1,2, Shouichi Okamoto1,2, Hiroki Ebana1,2,6,7, Yoshinori Okada8, Masatoshi Kurihara2,6, Kenji Suzuki9, Johji Inazawa5, Kazuhisa Takahashi1, Tetsuro Watabe3 & Kuniaki Seyama1,2 Lymphangioleiomyomatosis (LAM) is a rare pulmonary disease characterised by the proliferation of smooth muscle-like cells (LAM cells), and an abundance of lymphatic vessels in LAM lesions. Studies reported that vascular endothelial growth factor-D (VEGF-D) secreted by LAM cells contributes to LAM-associated lymphangiogenesis, however, the precise mechanisms of lymphangiogenesis and characteristics of lymphatic endothelial cells (LECs) in LAM lesions have not yet been elucidated. In this study, human primary-cultured LECs were obtained both from LAM-afected lung tissues (LAM-LECs) and normal lung tissues (control LECs) using fuorescence-activated cell sorting (FACS). We found that LAM-LECs had signifcantly higher ability of proliferation and migration compared to control LECs. VEGF-D signifcantly promoted migration of LECs but not proliferation of LECs in vitro. cDNA microarray and FACS analysis revealed the expression of vascular endothelial growth factor receptor (VEGFR)-3 and integrin α9 were elevated in LAM-LECs. Inhibition of VEGFR-3 suppressed proliferation and migration of LECs, and blockade of integrin α9 reduced VEGF-D-induced migration of LECs. Our data uncovered the distinct features of LAM-associated LECs, increased proliferation and migration, which may be due to higher expression of VEGFR-3 and integrin α9. Furthermore, we also found VEGF-D/VEGFR-3 and VEGF-D/ integrin α9 signaling play an important role in LAM-associated lymphangiogenesis.
    [Show full text]
  • The First Genome-Wide View of Vitamin D Receptor Locations and Their Mechanistic Implications
    ANTICANCER RESEARCH 32: 271-282 (2012) Review The First Genome-wide View of Vitamin D Receptor Locations and Their Mechanistic Implications CARSTEN CARLBERG1, SABINE SEUTER2 and SAMI HEIKKINEN1 1Department of Biosciences, University of Eastern Finland, Kuopio, Finland; 2Life Sciences Research Unit, University of Luxembourg, Luxembourg, Luxembourg Abstract. The transcription factor vitamin D receptor Physiological Impact of Vitamin D (VDR) is the nuclear sensor for the biologically most active in the Immune System metabolite of vitamin D, 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3). The physiological actions of the VDR and Vitamin D is a micronutrient which under ultraviolet (UV) its ligand are not only the well-known regulation of calcium radiation can also be produced in the skin (1). The most and phosphorus uptake and transport controlling bone abundant form of vitamin D is its liver hydroxylation product formation, but also their significant involvement in the 25-hydroxyvitamin D3 (25(OH)D3), serum concentrations of control of immune functions and of cellular growth and which indicate the vitamin D status of the human individual differentiation. For a general understanding of the (2). The biologically most active vitamin D metabolite is mechanisms of 1α,25(OH)2D3 signaling, it is essential to obtained from further hydroxylation of 25(OH)D3 in the monitor the genome-wide location of VDR in relation to kidney to 1α,25(OH)2D3 (3). Interestingly, the hydroxylation primary 1α,25(OH)2D3 target genes. Within the last months, of vitamin D can also take place in other tissues and a few of two chromatin immunoprecipitation sequencing (ChIP-Seq) them, such as keratinocytes and macrophages, have the studies using cells of the hematopoietic system, capacity for the full conversion of vitamin D to lymphoblastoids and monocytes, were published.
    [Show full text]
  • Interrelationships of Vitamin D and Parathyroid Hormone in Calcium Homeostasis Michael T
    POSTGRAD. MED. J. (1964), 40, 497 Postgrad Med J: first published as 10.1136/pgmj.40.466.497 on 1 August 1964. Downloaded from INTERRELATIONSHIPS OF VITAMIN D AND PARATHYROID HORMONE IN CALCIUM HOMEOSTASIS MICHAEL T. HARRISON, M.D., M.R.C.P. Lecturer in Medicine, University of Glasgow. From ihe Gardiner Institute of Medicine, Western Infirmary, Glasgow. IN man and most mammals the concentration and showed that reciprocal changes in con- of calcium in the blood and body fluids remains centration of calcium in the systemic circula- almost constant under normal conditions, with tion resulted. The mechanism by which the only slight fluctuations. For the various cellular parathyroid glands regulate their secretion in functions which are dependent on calcium, for response to changing levels of serum calcium example neuromuscular transmission, it is the is quite unknown. ionized fraction of calcium (approximately The parathyroid hormone has been puri- 50 per cent of the total serum calcium level) fied from bovine glands, and its properties which is important, and it is this fraction which have been studied (Rasmussen and Craig is so carefully maintained at a constant level 1961, 1962). It is a protein of molecular weight in the body fluids. In order to stabilize this approximately 9,000, composed of a single level, an elaborate regulating mechanism is chain containing 76 to 83 amino-acid residues. necessary, since there are many routes of entry It is antigenically pure, and antibodies have Protected by copyright. into and exit from the blood available to been prepared to it which provide the basis calcium.
    [Show full text]
  • Vitamin D and Growth Hormone in Children
    Esposito et al. J Transl Med (2019) 17:87 https://doi.org/10.1186/s12967-019-1840-4 Journal of Translational Medicine REVIEW Open Access Vitamin D and growth hormone in children: a review of the current scientifc knowledge Susanna Esposito* , Alberto Leonardi, Lucia Lanciotti, Marta Cofni, Giulia Muzi and Laura Penta Abstract Background: Human growth is a complex mechanism that depends on genetic, environmental, nutritional and hormonal factors. The main hormone involved in growth at each stage of development is growth hormone (GH) and its mediator, insulin-like growth factor 1 (IGF-1). In contrast, vitamin D is involved in the processes of bone growth and mineralization through the regulation of calcium and phosphorus metabolism. Nevertheless, no scientifc study has yet elucidated how they interact with one another, especially as a dysfunction in which one infuences the other, even if numerous biochemical and clinical studies confrm the presence of a close relationship. Main body: We reviewed and analyzed the clinical studies that have considered the relationship between vitamin D and the GH/IGF-1 axis in pediatric populations. We found two main areas of interest: the vitamin D defciency status in patients afected by GH defcit (GHD) and the relationship between serum vitamin D metabolites and IGF-1. Although limited by some bias, from the analysis of the studies presented in the scientifc literature, it is possible to hypothesize a greater frequency of hypovitaminosis D in the subjects afected by GHD, a reduced possibility of its correction with only substitution treatment with recombinant growth hormone (rGH) and an improvement of IGF-1 levels after sup- plementation treatment with vitamin D.
    [Show full text]
  • LSD1 Dual Function in Mediating Epigenetic Corruption of the Vitamin
    Battaglia et al. Clinical Epigenetics (2017) 9:82 DOI 10.1186/s13148-017-0382-y RESEARCH Open Access LSD1 dual function in mediating epigenetic corruption of the vitamin D signaling in prostate cancer Sebastiano Battaglia1*, Ellen Karasik2, Bryan Gillard2, Jennifer Williams2, Trisha Winchester3, Michael T. Moser2, Dominic J Smiraglia3 and Barbara A. Foster2* Abstract Background: Lysine-specific demethylase 1A (LSD1) is a key regulator of the androgen (AR) and estrogen receptors (ER), and LSD1 levels correlate with tumor aggressiveness. Here, we demonstrate that LSD1 regulates vitamin D receptor (VDR) activity and is a mediator of 1,25(OH)2-D3 (vitamin D) action in prostate cancer (PCa). Methods: Athymic nude mice were xenografted with CWR22 cells and monitored weekly after testosterone pellet removal. Expression of LSD1 and VDR (IHC) were correlated with tumor growth using log-rank test. TRAMP tumors and prostates from wild-type (WT) mice were used to evaluate VDR and LSD1 expression via IHC and western blotting. The presence of VDR and LSD1 in the same transcriptional complex was evaluated via immunoprecipitation (IP) using nuclear cell lysate. The effect of LSD1 and 1,25(OH)2-D3 on cell viability was evaluated in C4-2 and BC1A cells via trypanblueexclusion.TheroleofLSD1inVDR-mediatedgenetranscriptionwasevaluatedforCdkn1a, E2f1, Cyp24a1,andS100g via qRT-PCR-TaqMan and via chromatin immunoprecipitation assay. Methylation of Cdkn1a TSS was measured via bisulfite sequencing, and methylation of a panel of cancer-related genes was quantified using methyl arrays. The Cancer Genome Atlas data were retrieved to identify genes whose status correlates with LSD1 and DNA methyltransferase 1 (DNMT1).
    [Show full text]
  • Beyond Proteinuria: VDR Activation Reduces Renal Inflammation in Experimental Diabetic Nephropathy
    Am J Physiol Renal Physiol 302: F647–F657, 2012. First published December 14, 2011; doi:10.1152/ajprenal.00090.2011. Translational Physiology Beyond proteinuria: VDR activation reduces renal inflammation in experimental diabetic nephropathy Maria-Dolores Sanchez-Niño,3 Milica Bozic,1 Elizabeth Córdoba-Lanús,4 Petya Valcheva,1 Olga Gracia,1 Merce Ibarz,2 Elvira Fernandez,1 Juan F. Navarro-Gonzalez,4* Alberto Ortiz,3* and Jose Manuel Valdivielso1* 1Research Laboratory and Nephrology Department, 2Biochemistry Department, Hospital Universitari Arnau de Vilanova, IRBLLEIDA, Lleida; 3Nefrología, IIS-Fundación Jiménez Díaz, Universidad Autonoma de Madrid and Instituto Reina Sofia de Investigaciones Nefrológicas-IRSIN, Madrid; and 4Servicio de Nefrología y Unidad de Investigación, Hospital Universitario Nuestra Señora de Candelaria, Santa Cruz de Tenerife, Spain Submitted 8 February 2011; accepted in final form 7 December 2011 Sanchez-Niño M, Bozic M, Córdoba-Lanús E, Valcheva P, eventual decrease in GFR. Damage to highly differentiated Gracia O, Ibarz M, Fernandez E, Navarro-Gonzalez JF, Ortiz A, glomerular podocytes is thought to be an early event in DN Valdivielso JM. Beyond proteinuria: VDR activation reduces renal (35). In recent years, a direct association between inflammatory inflammation in experimental diabetic nephropathy. Am J Physiol parameters and clinical markers of glomerular as well as Renal Physiol 302: F647–F657, 2012. First published December 14, tubulointerstitial damage has been demonstrated, suggesting 2011; doi:10.1152/ajprenal.00090.2011.—Local inflammation is thought to contribute to the progression of diabetic nephropathy. The vitamin D that inflammation may be a pathogenic factor for the develop- ment and progression of DN (22). Hyperglycemia, angiotensin receptor (VDR) activator paricalcitol has an antiproteinuric effect in ␤ human diabetic nephropathy at high doses.
    [Show full text]
  • Vitamin D, Skeletal Muscle Function and Athletic Performance in Athletes—A Narrative Review
    nutrients Communication Vitamin D, Skeletal Muscle Function and Athletic Performance in Athletes—A Narrative Review Anna Ksi ˛a˙zek* , Aleksandra Zagrodna and Małgorzata Słowi ´nska-Lisowska Department of the Biological and Motor Basis of Sport, University School of Physical Education, Wrocław 51612, Poland * Correspondence: [email protected]; Tel.: +48-71-347-35-63 Received: 24 June 2019; Accepted: 2 August 2019; Published: 4 August 2019 Abstract: The active form of vitamin D (calcitriol) exerts its biological effects by binding to nuclear vitamin D receptors (VDRs), which are found in most human extraskeletal cells, including skeletal muscles. Vitamin D deficiency may cause deficits in strength, and lead to fatty degeneration of type II muscle fibers, which has been found to negatively correlate with physical performance. Vitamin D supplementation has been shown to improve vitamin D status and can positively affect skeletal muscles. The purpose of this study is to summarize the current evidence of the relationship between vitamin D, skeletal muscle function and physical performance in athletes. Additionally, we will discuss the effect of vitamin D supplementation on athletic performance in players. Further studies are necessary to fully characterize the underlying mechanisms of calcitriol action in the human skeletal muscle tissue, and to understand how these actions impact the athletic performance in athletes. Keywords: 25(OH)D; calcidiol; calcitriol; muscle performance; muscle strength; physical activity; athlete 1. Introduction Recent years have seen an increased interest in the research studies investigating vitamin D status in athletes. The growing number of scientific reports suggests a pleiotropic nature of vitamin D, suggested by the demonstration of vitamin D receptors (VDRs) nearly in every nucleated cell of our bodies.
    [Show full text]
  • 1Α,25‑Dihydroxyvitamin D3 Restrains Stem Cell‑Like Properties of Ovarian Cancer Cells by Enhancing Vitamin D Receptor and Suppressing CD44
    ONCOLOGY REPORTS 41: 3393-3403, 2019 1α,25‑Dihydroxyvitamin D3 restrains stem cell‑like properties of ovarian cancer cells by enhancing vitamin D receptor and suppressing CD44 MINTAO JI1*, LIZHI LIU2*, YONGFENG HOU3 and BINGYAN LI1 1Department of Nutrition and Food Hygiene, Soochow University of Public Health, Suzhou, Jiangsu 215123; 2State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat‑Sen University Cancer Center, Guangzhou, Guangdong 510060; 3State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, P.R. China Received May 10, 2018; Accepted February 27, 2019 DOI: 10.3892/or.2019.7116 Abstract. Scientific evidence linking vitamin D with various the expression of CD44. These findings provide a novel insight cancer types is growing, but the effects of vitamin D on ovarian into the functions of vitamin D in diminishing the stemness of cancer stem cell‑like cells (CSCs) are largely unknown. The cancer CSCs. present study aimed to examine whether vitamin D was able to restrain the stemness of ovarian cancer. A side popula- Introduction tion (SP) from malignant ovarian surface epithelial cells was identified as CSCs, in vitro and in vivo. Furthermore, Ovarian cancer is the fourth most frequent gynecologic malig- 1α,25‑dihydroxyvitamin D3 [1α,25(OH)2D3] treatment nancy and the leading cause of tumor‑associated mortality inhibited the self‑renewal capacity of SP cells by decreasing in the USA (1). Epithelial ovarian cancer, which accounts the sphere formation rate and by suppressing the mRNA for ~90% of ovarian cancers, is generally diagnosed at an expression levels of cluster of differentiation CD44, NANOG, advanced stage (2).
    [Show full text]