DOCSLIB.ORG

Agonist and Antagonist of Retinoic Acid Receptors Cause Similar Changes in Gene Expression and Induce Senescence-Like Growth Arrest in MCF-7 Breast Carcinoma Cells

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Agonist and Antagonist of Retinoic Acid Receptors Cause Similar Changes in Gene Expression and Induce Senescence-Like Growth Arrest in MCF-7 Breast Carcinoma Cells Research Article Agonist and Antagonist of Retinoic Acid Receptors Cause Similar Changes in Gene Expression and Induce Senescence-like Growth Arrest in MCF-7 Breast Carcinoma Cells Yuhong Chen,1 Milos Dokmanovic,1 Wilfred D. Stein,1,2 Robert J. Ardecky,3 and Igor B. Roninson1 1Cancer Center, Ordway Research Institute, Albany, New York; 2Institute of Life Sciences, Hebrew University, Jerusalem, Israel; and 3Ligand Pharmaceuticals, Inc., San Diego, California Abstract retinoids is most often attributed to the induction of differentia- Biological effects of retinoids are mediated via retinoic acid tion, but these compounds were also shown to stop the growth of (RA) receptors (RAR) and retinoid X receptors (RXR). The tumor cells by inducing apoptosis or accelerated senescence (1, 2). best-characterized mechanism of retinoid action is stimula- In particular, treatment of two human breast carcinoma cell lines tion of transcription from promoters containing RA response with all-trans retinoic acid (RA) or fenretinide, in vitro or in vivo, elements (RARE). Retinoids induce senescence-like growth induces a senescence-like phenotype characterized by increased h arrest in MCF-7 breast carcinoma cells; this effect is cell size and expression of senescence-associated -galactosidase h associated with the induction of several growth-inhibitory (SA- -gal; refs. 3, 4). This phenotype, as investigated in MCF-7 cells, genes. We have nowfound that these genes are induced by is associated with irreversible growth arrest and up-regulation of RAR-specific but not by RXR-specific ligands. Genome-scale several intracellular and secreted proteins with known growth- microarray analysis of gene expression was used to compare inhibitory activities. These include intracellular growth-inhibitory the effects of two pan-RAR ligands, one of which is a strong proteins, such as UBD (also known as FAT10) and putative tumor agonist of RARE-dependent transcription, whereas the other suppressor EPLIN, as well as secreted growth-inhibitory factors, induces such transcription only weakly and antagonizes the including insulin-like growth factor-binding protein 3 (IGFBP3) inducing effect of RAR agonists. Both RAR ligands, however, and an extracellular matrix component, TGFBI also known as h produced very similar effects on gene expression in MCF-7 IG-h3 (ref. 4). cells, suggesting that RARE-dependent transcription is only a Induction of gene expression by retinoids is mediated at the level minor component of retinoid-induced changes in gene of transcription through binding to dimeric transcription factors expression. The effects of RAR ligands on gene expression formed by RA receptors (RAR) and retinoid X receptors (RXR). The parallel changes associated with damage-induced senescence, best-known mechanism of action of these receptors involves their binding to RA response elements (RARE) in the promoters of andbothligandsinducedG1 arrest and the senescent phenotype in MCF-7 cells. The RAR ligands up-regulated retinoid-responsive genes. Nevertheless, retinoid receptors also many tumor-suppressive genes and down-regulated multiple affect transcription through RARE-independent mechanisms, such genes with oncogenic activities. Genes that are strongly as repression of transcription factor activator protein (AP-1; Jun/ induced by RAR ligands encode secreted bioactive proteins, Fos; ref. 5), or by modulating the interaction of Sp1 and GC-rich including several tumor-suppressing factors. In agreement DNA via ternary complex formation (6). Remarkably, a survey of with these observations, retinoid-treated MCF-7 cells inhibited Balmer and Blomhoff (7) concluded that only a minority of all the the growth of retinoid-insensitive MDA-MB-231 breast published retinoid-inducible genes are induced through the RARE- carcinoma cells in coculture. These results indicate that dependent mechanism. In the case of retinoid-treated MCF-7 cells, RARE-independent transcriptional effects of RAR ligands lead only 1 of 13 genes found to be strongly up-regulated at the onset of to senescence-like growth arrest and paracrine growth- senescence-like growth arrest contained a putative RARE sequence inhibitory activity in MCF-7 breast carcinoma cells. (Cancer in its promoter, whereas the other genes had no identifiable RARE Res 2006; 66(17): 8749-61) sites and showed a slow kinetics of retinoid response, requiring up to 3 days for maximal induction (4). Such genes may be induced Introduction either by an entirely RARE-independent mechanism or as a secondary consequence of some early RARE-dependent changes Retinoids, natural and synthetic derivatives of vitamin A, in gene expression. regulate growth, differentiation, and survival of different types of In the present study, we have used pan-RAR– and pan-RXR– normal and tumor cells. Retinoids are used in the treatment of specific agonists and antagonists to investigate the roles of promyelocytic leukemia and in chemoprevention of several retinoid receptors in the induction of growth-inhibitory genes in cancers, including breast carcinoma. The antitumor effect of MCF-7 cells. We have found that these genes are induced by the agonist of RAR (but nor RXR) and, surprisingly, by an RAR ligand that was developed as an antagonist of RARE-dependent Note: Current address for Milos Dokmanovic: Memorial Sloan-Kettering Cancer transcriptional activation. Microarray analysis of gene expression Center, Cell Biology Program, Sloan-Kettering Institute for Cancer Research, New York, showed that the agonist and the antagonist of RARE-dependent NY 10021. Requests for reprints: Igor B. Roninson, Cancer Center, Ordway Research transcription produced very similar effects on global gene Institute, 150 New Scotland Avenue, Albany, NY 12208. Phone: 518-641-6471; Fax: 518- expression in MCF-7 cells. In agreement with these effects, both 641-6305; E-mail: [email protected]. I2006 American Association for Cancer Research. RAR agonist and RAR antagonist induced senescence-like growth doi:10.1158/0008-5472.CAN-06-0581 arrest in MCF-7 cells. We have also identified numerous genes with www.aacrjournals.org 8749 Cancer Res 2006; 66: (17). September 1, 2006 Downloaded from cancerres.aacrjournals.org on September 30, 2021. © 2006 American Association for Cancer Research. Cancer Research oncogenic or tumor-suppressive properties that are affected by retinoid-treated MCF-7 cells, we have analyzed the effects of pan- RAR ligands and shown that the up-regulation of secreted tumor- RAR– and pan-RXR–specific agonists and antagonists on these cells. suppressive factors in retinoid-treated MCF-7 cells is associated The tested compounds include LGD1550, a pan-RAR agonist that with paracrine growth-inhibitory effect on retinoid-insensitive shows greater selectivity for RAR than natural retinoids (10); breast carcinoma cells. These results indicate that retinoids inhibit LGD1268 (a pan-RXR agonist); LG100815 (a pan-RAR antagonist); MCF-7 cell growth primarily through RARE-independent effects on and LG101208 (a pan-RXR antagonist). We have also used all-trans- cellular gene expression. RA, a natural RAR agonist that was originally used to define retinoid- induced senescence and changes in gene expression in MCF-7 cells Materials and Methods (3, 4). The structures of these compounds are shown in Fig. 1. In the first set of experiments, we asked how these compounds Cellular assays. MCF-7 cells were obtained from American Type Culture affect the expression of several growth-inhibitory genes previously Collection (Manassas, VA) and cultured in DMEM with 10% fetal bovine found to be strongly up-regulated under the conditions of retinoid- serum. Cells were plated at 105 per P100 (for assays requiring up to 6 days of 4 induced senescence of MCF-7 cells. The tested genes included culture) or at 10 per P60 (for longer assays), in the presence of different b concentrations of all-trans-RA (from Sigma, St. Louis, MO), pan-RAR EPLIN- , UBD (also known as FAT10), IGFBP3, and TGFBI (also b agonist LGD1550, pan-RXR agonist LGD1268, pan-RAR antagonist known as IG-h3), as well as TRIM31, a gene strongly induced by LG100815 or pan-RXR antagonist LG101208 (Ligand Pharmaceuticals, RA that has a putative RARE element in its promoter (4). RNA was Inc., San Diego, CA), or DMSO carrier. Following treatment, the number extracted from MCF-7 cells that were either untreated or treated of attached cells was measured using Coulter counter, and staining for with individual compounds or their combinations for 3 days, the SA-h-gal was carried out as described (8). For cell cycle analysis, cells were period previously shown to be required for the maximal effect of stained with propidium iodide by standard procedures. Cellular DNA RA or fenretinide (4). Gene expression was analyzed both by content was determined by flow cytometry using BD LSRII fluorescence- semiquantitative RT-PCR (not shown) and by QPCR; both assays activated cell sorting, and the percentages of cells in G1,S,orG2-M were produced essentially the same results. The outcome of a determined using ModFit software. representative set of QPCR assays is shown in Table 1. For coculture assays, MDA-MB-231 cells that were modified to express green fluorescent protein (GFP) from lentiviral vector pLL3.7 (9) were plated In agreement with the previous study, all five of the tested genes either alone (at 2 Â 105 per P100) or mixed with MCF-7 (at 105 cells from were induced by 100 nmol/L RA, the concentration found to be each cell line). In some assays, MCF-7 cells pretreated with 100 nmol/L RA sufficient for maximal induction of gene expression (4), with for 8 days were collected by trypsinization before mixing with MDA-MB- TRIM31 showing greater fold induction than the other four genes. 231. After culture in the presence or in the absence of 100 nmol/L RA, total The pan-RAR agonist LGD1550 induced all five genes as strongly as cell number was counted and the percentage of MDA-MB-231 GFP cells was RA; a 100 nmol/L concentration of this agonist was also sufficient determined by flow cytometry.
Load more

Recommended publications
  • Endothelial Cells and the IGF System
    L A Bach Endothelial cells as IGF targets 54:1 R1–R13 Review Endothelial cells and the IGF system Correspondence 1,2 Leon A Bach should be addressed to L A Bach 1Department of Medicine (Alfred), Monash University, Prahran 3181, Australia Email 2Department of Endocrinology and Diabetes, Alfred Hospital, Commercial Road, Melbourne 3004, Australia [email protected] Abstract Endothelial cells line blood vessels and modulate vascular tone, thrombosis, inflammatory Key Words responses and new vessel formation. They are implicated in many disease processes including " insulin-like growth factor atherosclerosis and cancer. IGFs play a significant role in the physiology of endothelial cells " binding protein by promoting migration, tube formation and production of the vasodilator nitric oxide. " receptor These actions are mediated by the IGF1 and IGF2/mannose 6-phosphate receptors and are " endothelial cell modulated by a family of high-affinity IGF binding proteins. IGFs also increase the number " angiogenesis and function of endothelial progenitor cells, which may contribute to protection from atherosclerosis. IGFs promote angiogenesis, and dysregulation of the IGF system may contribute to this process in cancer and eye diseases including retinopathy of prematurity and diabetic retinopathy. In some situations, IGF deficiency appears to contribute to endothelial dysfunction, whereas IGF may be deleterious in others. These differences may be due to tissue-specific endothelial cell phenotypes or IGFs having distinct roles in different phases of vascular disease. Further studies are therefore required to delineate the Journal of Molecular therapeutic potential of IGF system modulation in pathogenic processes. Endocrinology (2015) 54, R1–R13 Journal of Molecular Endocrinology Introduction Insulin-like growth factor 1 (IGF1) and IGF2 are essential metabolically active and regulate vascular tone, thrombosis, for normal pre- and postnatal growth and development inflammatory responses and new vessel formation.
    [Show full text]
  • 2017.08.28 Anne Barry-Reidy Thesis Final.Pdf
    REGULATION OF BOVINE β-DEFENSIN EXPRESSION THIS THESIS IS SUBMITTED TO THE UNIVERSITY OF DUBLIN FOR THE DEGREE OF DOCTOR OF PHILOSOPHY 2017 ANNE BARRY-REIDY SCHOOL OF BIOCHEMISTRY & IMMUNOLOGY TRINITY COLLEGE DUBLIN SUPERVISORS: PROF. CLIONA O’FARRELLY & DR. KIERAN MEADE TABLE OF CONTENTS DECLARATION ................................................................................................................................. vii ACKNOWLEDGEMENTS ................................................................................................................... viii ABBREVIATIONS ................................................................................................................................ix LIST OF FIGURES............................................................................................................................. xiii LIST OF TABLES .............................................................................................................................. xvii ABSTRACT ........................................................................................................................................xix Chapter 1 Introduction ........................................................................................................ 1 1.1 Antimicrobial/Host-defence peptides ..................................................................... 1 1.2 Defensins................................................................................................................. 1 1.3 β-defensins .............................................................................................................
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Farnesol-Induced Apoptosis in Human Lung Carcinoma Cells Is Coupled to the Endoplasmic Reticulum Stress Response
    Research Article Farnesol-Induced Apoptosis in Human Lung Carcinoma Cells Is Coupled to the Endoplasmic Reticulum Stress Response Joung Hyuck Joo,1 Grace Liao,1 Jennifer B. Collins,2 Sherry F. Grissom,2 and Anton M. Jetten1 1Cell Biology Section, LRB, and 2Microarray Group, Division of Intramural Research, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, North Carolina Abstract range of fruits and vegetables (9, 10). Each isoprenoid has been Farnesol (FOH) and other isoprenoid alcohols induce apopto- shown to inhibit proliferation and induce apoptosis in a number of sis in various carcinoma cells and inhibit tumorigenesis in neoplastic cell lines from different origins (4, 11–14). In addition, in vivo these isoprenoids have been reported to be effective in chemo- several models. However, the mechanisms by which in vivo they mediate their effects are not yet fully understood. In this prevention and chemotherapy in various cancer models study, we show that FOH is an effective inducer of apoptosis in (10, 12, 15, 16). FOH has been reported to exhibit chemopreventive several lung carcinoma cells, including H460. This induction is effects in colon and pancreas carcinogenesis in rats (9, 17) whereas associated with activation of several caspases and cleavage of phase I and II clinical trials have indicated therapeutic potential poly(ADP-ribose) polymerase (PARP). To obtain insight into for POH (16, 18). The mechanisms by which these isoprenoids induce these effects are not yet fully understood. Isoprenoids have the mechanism involved in FOH-induced apoptosis, we compared the gene expression profiles of FOH-treated and been reported to inhibit posttranslational protein prenylation (19) control H460 cells by microarray analysis.
    [Show full text]
  • Partial Deletion of Chromosome 6P Causing Developmental Delay and Mild Dysmorphisms in a Child
    Vrachnis et al. Mol Cytogenet (2021) 14:39 https://doi.org/10.1186/s13039-021-00557-y CASE REPORT Open Access Partial deletion of chromosome 6p causing developmental delay and mild dysmorphisms in a child: molecular and developmental investigation and literature search Nikolaos Vrachnis1,2,3* , Ioannis Papoulidis4, Dionysios Vrachnis5, Elisavet Siomou4, Nikolaos Antonakopoulos1,2, Stavroula Oikonomou6, Dimitrios Zygouris2, Nikolaos Loukas7, Zoi Iliodromiti8, Efterpi Pavlidou9, Loretta Thomaidis6 and Emmanouil Manolakos4 Abstract Background: The interstitial 6p22.3 deletions concern rare chromosomal events afecting numerous aspects of both physical and mental development. The syndrome is characterized by partial deletion of chromosome 6, which may arise in a number of ways. Case presentation: We report a 2.8-year old boy presenting with developmental delay and mild dysmorphisms. High-resolution oligonucleotide microarray analysis revealed with high precision a 2.5 Mb interstitial 6p deletion in the 6p22.3 region which encompasses 13 genes. Conclusions: Identifcation and in-depth analysis of cases presenting with mild features of the syndrome will sharpen our understanding of the genetic spectrum of the 6p22.3 deletion. Keywords: 6p22.3 deletion, Syndrome, Developmental delay, Intellectual disability, Dysmorphism, Behavioral abnormalities, High-resolution microarray analysis Background dysmorphic features, and structural organ defects, as well Te interstitial deletion of chromosomal region 6p22.3 as intellectual disability. is a rare condition with variable phenotypic expression. We report herein a case of interstitial deletion of chro- To date, more than 30 children and adolescents with this mosome 6p investigated by array-CGH in a 2.8-year old deletion have been reported [1–11].
    [Show full text]
  • Temporal Chip-On-Chip of RNA-Polymerase-II to Detect Novel Gene Activation Events During Photoreceptor Maturation
    Molecular Vision 2010; 16:252-271 <http://www.molvis.org/molvis/v16/a32> © 2010 Molecular Vision Received 12 July 2009 | Accepted 10 February 2010 | Published 17 February 2010 Temporal ChIP-on-Chip of RNA-Polymerase-II to detect novel gene activation events during photoreceptor maturation Padmaja Tummala, Raghuveer S. Mali, Eduardo Guzman, Xiao Zhang, Kenneth P. Mitton (The first two authors contributed equally to this work.) Eye Research Institute, Oakland University, Rochester, MI Purpose: During retinal development, post-mitotic neural progenitor cells must activate thousands of genes to complete synaptogenesis and terminal maturation. While many of these genes are known, others remain beyond the sensitivity of expression microarray analysis. Some of these elusive gene activation events can be detected by mapping changes in RNA polymerase-II (Pol-II) association around transcription start sites. Methods: High-resolution (35 bp) chromatin immunoprecipitation (ChIP)-on-chip was used to map changes in Pol-II binding surrounding 26,000 gene transcription start sites during photoreceptor maturation of the mouse neural retina, comparing postnatal age 25 (P25) to P2. Coverage was 10–12 kb per transcription start site, including 2.5 kb downstream. Pol-II-active regions were mapped to the mouse genomic DNA sequence by using computational methods (Tiling Analysis Software-TAS program), and the ratio of maximum Pol-II binding (P25/P2) was calculated for each gene. A validation set of 36 genes (3%), representing a full range of Pol-II signal ratios (P25/P2), were examined with quantitative ChIP assays for transcriptionally active Pol-II. Gene expression assays were also performed for 19 genes of the validation set, again on independent samples.
    [Show full text]
  • 4-Hydroxyphenyiretinamide
    Handbook 4 4-Hydroxyphenyiretinamide 1. Chemical and Physical Melting-point Characteristics 173-175 °C (Shealy etal., 1984; Budavari etal., 1996) 1.1. Nomenclature Solubility See General Remarks, section 1.4. Soluble in most organic solvents, fats, oils and aqueous micellar solutions; sparingly soluble in 1.2 Name water, e.g. 13 nmol/L at pH 6.5 (Li et al., 1996). Chemical Abstracts Services Registry Number 65646-68-6 Spectroscopy 1% UV and visible: ? 362 (methanol), E 1 IUPAC systematic name 1225, EM 47 900 (Budavari et al., 1996; Barua & N-[4-(all-E)-9, 13-Dimethyl-7-(1, 1,5-trimethylcy- Fun, 1998). Higher EM values, i.e. 57 100 and 56 clohex-5 -en-6-yl)nona- 7,9,11,1 3-tetraen- 15- 400, have also been reported (Formelli et al., 1996). oyl]aminophenol or N-[4-(all E)-3, 7-dimethyl-9- (2,2,6-trimethylcyclohex-1-en-1-yl)nona-2,4,6,8- Nuclear magnetic resonance: tetraen-1-oyl]aminophenol 'H-NMR [(CD3)SO4, 100 MHz]: 8 1.03 (1,1-CH3), 1.3-1.8 (2-CH2, 3-CH), 1.70 (5-CH), 1.8-2.2 Synonyms (4-CH2), 1.99 (9-CH), 2.36 (13-CH), 6.03 (14-H), Fenretinide, 4-HPR, N-(4-hydroxyphenyl)reti- 6.1-6.6 (7-H, 8-H, 10-H, 12-H), 6.99 (11-H), namide, hydroxyphenyl-retinamide, N-(4-hydro- 6.6-6.8 and 7.3-7.6 (benzene ring -H), 9.15 (NH), xyphenyl)retinamide, N-(4-hydroxyphenyl)-all- 9.74 (OH) (Coburn et al., 1983; Shealy et al., tnins-retinamide 1984).
    [Show full text]
  • Epigenome-Wide Exploratory Study of Monozygotic Twins Suggests Differentially Methylated Regions to Associate with Hand Grip Strength
    Biogerontology (2019) 20:627–647 https://doi.org/10.1007/s10522-019-09818-1 (0123456789().,-volV)( 0123456789().,-volV) RESEARCH ARTICLE Epigenome-wide exploratory study of monozygotic twins suggests differentially methylated regions to associate with hand grip strength Mette Soerensen . Weilong Li . Birgit Debrabant . Marianne Nygaard . Jonas Mengel-From . Morten Frost . Kaare Christensen . Lene Christiansen . Qihua Tan Received: 15 April 2019 / Accepted: 24 June 2019 / Published online: 28 June 2019 Ó The Author(s) 2019 Abstract Hand grip strength is a measure of mus- significant CpG sites or pathways were found, how- cular strength and is used to study age-related loss of ever two of the suggestive top CpG sites were mapped physical capacity. In order to explore the biological to the COL6A1 and CACNA1B genes, known to be mechanisms that influence hand grip strength varia- related to muscular dysfunction. By investigating tion, an epigenome-wide association study (EWAS) of genomic regions using the comb-p algorithm, several hand grip strength in 672 middle-aged and elderly differentially methylated regions in regulatory monozygotic twins (age 55–90 years) was performed, domains were identified as significantly associated to using both individual and twin pair level analyses, the hand grip strength, and pathway analyses of these latter controlling the influence of genetic variation. regions revealed significant pathways related to the Moreover, as measurements of hand grip strength immune system, autoimmune disorders, including performed over 8 years were available in the elderly diabetes type 1 and viral myocarditis, as well as twins (age 73–90 at intake), a longitudinal EWAS was negative regulation of cell differentiation.
    [Show full text]
  • SNORD116 and Growth Hormone Therapy Impact IGFBP7 in Praderâ
    www.nature.com/gim ARTICLE SNORD116 and growth hormone therapy impact IGFBP7 in Prader–Willi syndrome Sanaa Eddiry1,2, Gwenaelle Diene3,4, Catherine Molinas1,3,4, Juliette Salles1,5, Françoise Conte Auriol1,2, Isabelle Gennero1, Eric Bieth6, ✉ Boris V. Skryabin7, Timofey S. Rozhdestvensky7, Lisa C. Burnett8, Rudolph L. Leibel9, Maithé Tauber1,3,4 and Jean Pierre Salles 1,2,4 PURPOSE: Prader–Willi syndrome (PWS) is a neurodevelopmental disorder with hypothalamic dysfunction due to deficiency of imprinted genes located on the 15q11-q13 chromosome. Among them, the SNORD116 gene appears critical for the expression of the PWS phenotype. We aimed to clarify the role of SNORD116 in cellular and animal models with regard to growth hormone therapy (GHT), the main approved treatment for PWS. METHODS: We collected serum and induced pluripotent stem cells (iPSCs) from GH-treated PWS patients to differentiate into dopaminergic neurons, and in parallel used a Snord116 knockout mouse model. We analyzed the expression of factors potentially linked to GH responsiveness. RESULTS: We found elevated levels of circulating IGFBP7 in naive PWS patients, with IGFBP7 levels normalizing under GHT. We found elevated IGFBP7 levels in the brains of Snord116 knockout mice and in iPSC-derived neurons from a SNORD116-deleted PWS patient. High circulating levels of IGFBP7 in PWS patients may result from both increased IGFBP7 expression and decreased IGFBP7 cleavage, by downregulation of the proconvertase PC1. CONCLUSION: SNORD116 deletion affects IGFBP7 levels, while IGFBP7 decreases under GHT in PWS patients. Modulation of the 1234567890():,; IGFBP7 level, which interacts with IGF1, has implications in the pathophysiology and management of PWS under GHT.
    [Show full text]
  • Expression Signatures of the Lipid-Based Akt Inhibitors Phosphatidylinositol Ether Lipid Analogues in NSCLC Cells
    Published OnlineFirst May 6, 2011; DOI: 10.1158/1535-7163.MCT-10-1028 Molecular Cancer Therapeutic Discovery Therapeutics Expression Signatures of the Lipid-Based Akt Inhibitors Phosphatidylinositol Ether Lipid Analogues in NSCLC Cells Chunyu Zhang1, Abdel G. Elkahloun2, Hongling Liao3, Shannon Delaney1, Barbara Saber1, Betsy Morrow1, George C. Prendergast4, M. Christine Hollander1, Joell J. Gills1, and Phillip A. Dennis1 Abstract Activation of the serine/threonine kinase Akt contributes to the formation, maintenance, and therapeutic resistance of cancer, which is driving development of compounds that inhibit Akt. Phosphatidylinositol ether lipid analogues (PIA) are analogues of the products of phosphoinositide-3-kinase (PI3K) that inhibit Akt activation, translocation, and the proliferation of a broad spectrum of cancer cell types. To gain insight into the mechanism of PIAs, time-dependent transcriptional profiling of five active PIAs and the PI3K inhibitor LY294002 (LY) was conducted in non–small cell lung carcinoma cells using high-density oligonucleotide arrays. Gene ontology analysis revealed that genes involved in apoptosis, wounding response, and angiogen- esis were upregulated by PIAs, whereas genes involved in DNA replication, repair, and mitosis were suppressed. Genes that exhibited early differential expression were partitioned into three groups; those induced by PIAs only (DUSP1, KLF6, CENTD2, BHLHB2, and PREX1), those commonly induced by PIAs and LY (TRIB1, KLF2, RHOB, and CDKN1A), and those commonly suppressed by PIAs and LY (IGFBP3, PCNA, PRIM1, MCM3, and HSPA1B). Increased expression of the tumor suppressors RHOB (RhoB), KLF6 (COPEB), and CDKN1A (p21Cip1/Waf1) was validated as an Akt-independent effect that contributed to PIA-induced cytotoxicity. Despite some overlap with LY, active PIAs have a distinct expression signature that contributes to their enhanced cytotoxicity.
    [Show full text]
  • Novel Regulators of the IGF System in Cancer
    biomolecules Review Novel Regulators of the IGF System in Cancer Caterina Mancarella 1, Andrea Morrione 2 and Katia Scotlandi 1,* 1 IRCCS Istituto Ortopedico Rizzoli, Laboratory of Experimental Oncology, 40136 Bologna, Italy; [email protected] 2 Department of Biology, Sbarro Institute for Cancer Research and Molecular Medicine and Center for Biotechnology, College of Science and Technology, Temple University, Philadelphia, PA 19122, USA; [email protected] * Correspondence: [email protected]; Tel.: +39-051-6366-760 Abstract: The insulin-like growth factor (IGF) system is a dynamic network of proteins, which includes cognate ligands, membrane receptors, ligand binding proteins and functional downstream effectors. It plays a critical role in regulating several important physiological processes including cell growth, metabolism and differentiation. Importantly, alterations in expression levels or activa- tion of components of the IGF network are implicated in many pathological conditions including diabetes, obesity and cancer initiation and progression. In this review we will initially cover some general aspects of IGF action and regulation in cancer and then focus in particular on the role of transcriptional regulators and novel interacting proteins, which functionally contribute in fine tuning IGF1R signaling in several cancer models. A deeper understanding of the biological relevance of this network of IGF1R modulators might provide novel therapeutic opportunities to block this system in neoplasia. Keywords: IGF system; cancer; transcriptional regulators; functional regulation; circular RNAs; IGF2BPs; ADAR; DDR1; E-cadherin; decorin Citation: Mancarella, C.; Morrione, A.; Scotlandi, K. Novel Regulators of the IGF System in Cancer. 1. Introduction Biomolecules 2021, 11, 273. https:// doi.org/10.3390/biom11020273 The insulin-like growth factor (IGF) system is a network of ligands, binding proteins and receptors regulating crucial physiological and pathological biological processes.
    [Show full text]
  • IGFBP5) Reverses Cisplatin-Resistance in Esophageal Carcinoma
    cells Article Expression of Insulin-Like Growth Factor Binding Protein-5 (IGFBP5) Reverses Cisplatin-Resistance in Esophageal Carcinoma Dessy Chan 1,†, Yuanyuan Zhou 1,†, Chung Hin Chui 1, Kim Hung Lam 1, Simon Law 2, Albert Sun-chi Chan 3, Xingshu Li 3,*, Alfred King-yin Lam 4,* and Johnny Cheuk On Tang 1,* 1 State Key Laboratory of Chemical Biology and Drug Discovery, Lo Ka Chung Centre for Natural Anti-cancer Drug Development, Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hong Kong, China; [email protected] (D.C.); [email protected] (Y.Z.); [email protected] (C.H.C.), [email protected] (K.H.L.) 2 Department of Surgery, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China; [email protected] 3 School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China; [email protected] 4 Griffith Medical School, Griffith University, Gold Coast, QLD 4222, Australia * Correspondence: [email protected] (X.L.); A.Lam@griffith.edu.au (A.K.L.); [email protected] (J.C.O.T.); Tel.: +852-3400-8727 (J.C.O.T.) † These authors contributed equally to this work. Received: 3 September 2018; Accepted: 16 September 2018; Published: 20 September 2018 Abstract: Cisplatin (CDDP) is one of the front-line chemotherapeutic drugs used in the treatment of esophageal squamous cell carcinoma (ESCC). Occurrence of resistance to CDDP has become one of the main challenges in cancer therapy. In this study, the gene expression profile of CDDP-resistant ESCC cells was investigated and molecular approaches were explored in an attempt to reverse the CDDP resistance.
    [Show full text]