DOCSLIB.ORG

Prox1 Suppresses the Proliferation of Neuroblastoma Cells Via a Dual Action in P27-Kip1 and Cdc25a

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Prox1 Suppresses the Proliferation of Neuroblastoma Cells Via a Dual Action in P27-Kip1 and Cdc25a Oncogene (2013) 32, 947–960 & 2013 Macmillan Publishers Limited All rights reserved 0950-9232/13 www.nature.com/onc ORIGINAL ARTICLE Prox1 suppresses the proliferation of neuroblastoma cells via a dual action in p27-Kip1 and Cdc25A IP Foskolou, D Stellas, I Rozani, MD Lavigne and PK Politis Neuroblastoma is a pediatric tumor that originates from precursor cells of the sympathetic nervous system with less than 40% long- term survival in children diagnosed with high-risk disease. These clinical observations underscore the need for novel insights in the mechanisms of malignant transformation and progression. Accordingly, it was recently reported that Prox1, a homeobox transcription regulator, is expressed in higher levels in human neuroblastoma with favorable prognosis. Consistently, we have recently shown that Prox1 exerts a strong antiproliferative effect on neural precursor cells during embryonic development. Thus, Prox1 is a candidate gene with a critical role in suppressing malignant neuroblastoma transformation. Here, we provide evidence that Prox1 strongly suppresses the proliferation of mouse and human neuroblastoma cell lines and blocks the growth of neuroblastoma tumors in SCID mice. Conversely, short hairpin RNA (shRNA) -mediated knockdown of basal Prox1 expression significantly induces proliferation, genomic instability and the ability of neuroblastoma cells to form tumors. Mechanistically, analysis of an inducible Prox1-overexpressing Neuro2A cell line indicates that Prox1 is sufficient to suppress CyclinD1, CyclinA and CyclinB1, consistent with a role in cell cycle arrest. Surprisingly, Prox1 strongly induces CyclinE1 expression in the same system despite its action on blocking cell cycle progression, which could account for the context dependent oncogenic function of Prox1. Most importantly, Prox1 was sufficient to decrease Cdc25A and induce p27-Kip1, but not p21-Cip1 or p53. By alleviating the Prox1 action in Cdc25A and p27-Kip1 expression, we were able to rescue its effect on cell cycle arrest. Together these data suggest that Prox1 negatively regulates neuroblastoma carcinogenesis through suppression of Cdc25A and induction of p27-Kip1 to counteract CyclinE1 overexpression and block cell cycle progression. Furthermore, these observations render Prox1 a candidate target for the treatment of neuroblastoma tumors. Oncogene (2013) 32, 947–960; doi:10.1038/onc.2012.129; published online 16 April 2012 Keywords: prospero; cyclinE1; G0/G1-arrest; cell cycle; SH-SY5Y; tet-on INTRODUCTION of proliferation and differentiation in neural precursor cells.5 In Neuroblastoma is a malignancy of early childhood that arises from agreement, recent data suggest that Notch1 expression predicts the developing autonomic nervous system.1 The clinical behavior an unfavorable prognosis for patients with neuroblastoma and is variable, ranging from spontaneous regression to highly could serve as a therapeutic target.6 Therefore, Prox1 may be aggressive metastatic disease with a poor overall survival rate. It involved in the malignant transformation, progression and/or is the most frequently diagnosed neoplasm during infancy and regression of neuroblastoma through Notch signaling regulation less than 40% of children with high-risk neuroblastoma are likely and/or additional genes and pathways. to achieve long-term cure, despite dramatic escalations in the Multiple lines of evidence suggest important roles for Prox1 in intensity of therapy provided.2 Future therapies will focus different aspects of embryonic development and morphogenesis, increasingly on the genes and biological pathways that while mouse embryos deficient in Prox1 die at E14.5. Prox1 has contribute to this malignancy. been previously shown to have essential roles during lymphatic, Towards this aim, it was recently reported that expression of hepatocyte, pancreatic, heart, lens, retinal and spinal cord Prox1, a homeobox transcription regulator, in human neuroblas- development.5,7–10 Prox1 has also been implicated in both toma tumors is correlated with the stage 4S, which has favorable progression and suppression of malignancies. A function for prognosis with high incidence of spontaneous regression. More- Prox1 as tumor suppressor has been described previously in other over, it was shown that Prox1 is highly expressed in sympathetic systems. Decreased expression levels have been found in neurons during early stages of development in chick, mouse and hepatocellular and pancreatic carcinoma,11,12 which may be due human peripheral nervous system and its expression levels are to epigenetic silencing. Hypermethylation of the Prox1 gene has reduced in highly proliferative human neuroblastoma cell lines.3,4 been found in breast cancer and lymphomas and in brain These observations suggest that Prox1 might have a tumor metastases of breast cancer.13,14 Moreover, mutations of Prox1 suppressor function in neuroblastoma cancer. Accordingly, we and loss of heterozygosity have been reported in various have recently reported that Prox1 negatively regulates the cancers.13,15,16 Although most studies point to a tumor proliferation of neural precursor cells during embryonic suppressor function of Prox1, a recent study reports that it development. Furthermore, we showed that a cross-inhibitory enhances colorectal cancer progression.17 These data suggest that interaction between Prox1 and Notch1 is involved in the regulation Prox1 may function in a context dependent manner, being a Center for Basic Research, Biomedical Research Foundation of the Academy of Athens, Athens, Greece. Correspondence: Dr PK Politis, Center for Basic Research, Biomedical Research Foundation of the Academy of Athens, 4 Soranou Efesiou, Athens 11527, Greece. E-mail: [email protected] Received 1 November 2011; revised 28 February 2012; accepted 7 March 2012; published online 16 April 2012 Prox1 blocks cell cycle progression in neuroblastoma cells IP Foskolou et al 948 tumor suppressor in certain cells, while in other cell types could be RESULTS related to tumor progression. This dual function may reside in the Prox1 blocks proliferation of neuroblastoma cancer cells fact that Prox1 affects differentially the expression of genes that To evaluate whether the observed correlation between Prox1 promote or inhibit proliferation and cell cycle progression. expression and suppression of neuroblastoma progression has Here, we provide functional evidence that Prox1 is indeed functional importance, we used neuroblastoma cell lines as a involved in the suppression of neuroblastoma cell growth both model system to analyze proliferation. We first examined the in vitro and in vivo. Mechanistically, activated Notch1 signaling effect of mouse or human Prox1 overexpression in Neuro2A (N2A) cannot rescue the negative effect of Prox1 on neuroblastoma mouse neuroblastoma cell line. BrdU incorporation assays proliferation, suggesting an alternative mode of action. Accord- revealed a strong block in proliferation after mouse or human ingly, we showed that Prox1 acts in basic components of cell cycle Prox1 overexpression (Figures 1a and b and Supplementary Figure machinery to regulate cellular proliferation in neuroblastoma cells. S1a). In addition, immunostainings with phosphorylated-histo- Collectively, these observations indicate that Prox1 negatively neH3 (pH3) revealed a strong reduction in Prox1 þ N2A cells regulates tumorigenic properties of neuroblastoma cells, may be undergoing mitosis (Figures 1d and e and Supplementary Figure implicated in suppressing neuroblastoma carcinogenesis, and S1b). To exclude the possibility of non-cell autonomous effects, we could be utilized for its treatment. also measured the proliferation rate in the nontransfected cells. In Figure 1. Prox1 has a strong antiproliferative effect on mouse and human neuroblastoma cell lines. (a) Prox1 or GFP transfected N2A cells were pulsed for 2 h with BrdU and then labeled for BrdU and 4,6-diamidino-2-phenylindole (DAPI). Scale bar: 25 mm. (b) Quantification of the transgene-positive/BrdU-positive cells. (GFP: 43.6±4.26%, Prox1: 1.86±0.59%; Po0.01) (c) Quantification of the transgene negative/BrdU- positive cells (GFP: 40.56±3.01%, Prox1: 39.13±1.66%; P40.1). (d) Prox1- or GFP-transfected N2A cells were labeled for pH3 and DAPI. Scale bar: 25 mm. (e) Quantification of the transgene-positive/pH3-positive cells (GFP: 11.52±1.2%, Prox1: 1.09±0.25%; Po0.05). (f) Quantification of the transgene-negative/pH3-positive cells (GFP: 11.66±0.47%, Prox1: 11.47±1.2%; P40.1). (g, h) Quantification of the transgene-positive/ BrdU-positive cells in SH-SY5Y and Kelly human neuroblastoma cell lines, respectively, (GFP: 23.78±1.68%, Prox1: 3.89±0.59%; Po0.01 in SH-SY5Y cells; GFP: 51.37±5.29%; Prox1: 20.5±0.86%; Po0.05 in Kelly cells). Oncogene (2013) 947 – 960 & 2013 Macmillan Publishers Limited Prox1 blocks cell cycle progression in neuroblastoma cells IP Foskolou et al 949 all cases, the untransfected cells in Prox1 experiments showed observations with MTT assay and PCNA immunostainings (Figures proliferation rates similar to green fluorescent protein (GFP) 3k–m). Collectively, these observations show that Prox1 negatively experiments (Figures 1c and f), further confirming that the defect affects proliferative and tumor forming abilities of neuroblastoma in proliferation is because of Prox1 overexpression. Prox1 was also cells in vitro with no indication of increased cell death. sufficient to suppress proliferation in human neuroblastoma cell lines, including SH-SY5Y and Kelly (Figures
Load more

Recommended publications
  • In Vivo Studies Using the Classical Mouse Diversity Panel
    The Mouse Diversity Panel Predicts Clinical Drug Toxicity Risk Where Classical Models Fail Alison Harrill, Ph.D The Hamner-UNC Institute for Drug Safety Sciences 0 The Importance of Predicting Clinical Adverse Drug Reactions (ADR) Figure: Cath O’Driscoll Nature Publishing 2004 Risk ID PGx Testing 1 People Respond Differently to Drugs Pharmacogenetic Markers Identified by Genome-Wide Association Drug Adverse Drug Risk Allele Reaction (ADR) Abacavir Hypersensitivity HLA-B*5701 Flucloxacillin Hepatotoxicity Allopurinol Cutaneous ADR HLA-B*5801 Carbamazepine Stevens-Johnson HLA-B*1502 Syndrome Augmentin Hepatotoxicity DRB1*1501 Ximelagatran Hepatotoxicity DRB1*0701 Ticlopidine Hepatotoxicity HLA-A*3303 Average preclinical populations and human hepatocytes lack the diversity to detect incidence of adverse events that occur only in 1/10,000 people. Current Rodent Models of Risk Assessment The Challenge “At a time of extraordinary scientific progress, methods have hardly changed in several decades ([FDA] 2004)… Toxicologists face a major challenge in the twenty-first century. They need to embrace the new “omics” techniques and ensure that they are using the most appropriate animals if their discipline is to become a more effective tool in drug development.” -Dr. Michael Festing Quantitative geneticist Toxicol Pathol. 2010;38(5):681-90 Rodent Models as a Strategy for Hazard Characterization and Pharmacogenetics Genetically defined rodent models may provide ability to: 1. Improve preclinical prediction of drugs that carry a human safety risk 2.
    [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]
  • To Study Mutant P53 Gain of Function, Various Tumor-Derived P53 Mutants
    Differential effects of mutant TAp63γ on transactivation of p53 and/or p63 responsive genes and their effects on global gene expression. A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science By Shama K Khokhar M.Sc., Bilaspur University, 2004 B.Sc., Bhopal University, 2002 2007 1 COPYRIGHT SHAMA K KHOKHAR 2007 2 WRIGHT STATE UNIVERSITY SCHOOL OF GRADUATE STUDIES Date of Defense: 12-03-07 I HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER MY SUPERVISION BY SHAMA KHAN KHOKHAR ENTITLED Differential effects of mutant TAp63γ on transactivation of p53 and/or p63 responsive genes and their effects on global gene expression BE ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Master of Science Madhavi P. Kadakia, Ph.D. Thesis Director Daniel Organisciak , Ph.D. Department Chair Committee on Final Examination Madhavi P. Kadakia, Ph.D. Steven J. Berberich, Ph.D. Michael Leffak, Ph.D. Joseph F. Thomas, Jr., Ph.D. Dean, School of Graduate Studies 3 Abstract Khokhar, Shama K. M.S., Department of Biochemistry and Molecular Biology, Wright State University, 2007 Differential effect of TAp63γ mutants on transactivation of p53 and/or p63 responsive genes and their effects on global gene expression. p63, a member of the p53 gene family, known to play a role in development, has more recently also been implicated in cancer progression. Mice lacking p63 exhibit severe developmental defects such as limb truncations, abnormal skin, and absence of hair follicles, teeth, and mammary glands. Germline missense mutations of p63 have been shown to be responsible for several human developmental syndromes including SHFM, EEC and ADULT syndromes and are associated with anomalies in the development of organs of epithelial origin.
    [Show full text]
  • Prox1regulates the Subtype-Specific Development of Caudal Ganglionic
    The Journal of Neuroscience, September 16, 2015 • 35(37):12869–12889 • 12869 Development/Plasticity/Repair Prox1 Regulates the Subtype-Specific Development of Caudal Ganglionic Eminence-Derived GABAergic Cortical Interneurons X Goichi Miyoshi,1 Allison Young,1 Timothy Petros,1 Theofanis Karayannis,1 Melissa McKenzie Chang,1 Alfonso Lavado,2 Tomohiko Iwano,3 Miho Nakajima,4 Hiroki Taniguchi,5 Z. Josh Huang,5 XNathaniel Heintz,4 Guillermo Oliver,2 Fumio Matsuzaki,3 Robert P. Machold,1 and Gord Fishell1 1Department of Neuroscience and Physiology, NYU Neuroscience Institute, Smilow Research Center, New York University School of Medicine, New York, New York 10016, 2Department of Genetics & Tumor Cell Biology, St. Jude Children’s Research Hospital, Memphis, Tennessee 38105, 3Laboratory for Cell Asymmetry, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan, 4Laboratory of Molecular Biology, Howard Hughes Medical Institute, GENSAT Project, The Rockefeller University, New York, New York 10065, and 5Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 11724 Neurogliaform (RELNϩ) and bipolar (VIPϩ) GABAergic interneurons of the mammalian cerebral cortex provide critical inhibition locally within the superficial layers. While these subtypes are known to originate from the embryonic caudal ganglionic eminence (CGE), the specific genetic programs that direct their positioning, maturation, and integration into the cortical network have not been eluci- dated. Here, we report that in mice expression of the transcription factor Prox1 is selectively maintained in postmitotic CGE-derived cortical interneuron precursors and that loss of Prox1 impairs the integration of these cells into superficial layers. Moreover, Prox1 differentially regulates the postnatal maturation of each specific subtype originating from the CGE (RELN, Calb2/VIP, and VIP).
    [Show full text]
  • Mouse Population-Guided Resequencing Reveals That Variants in CD44 Contribute to Acetaminophen-Induced Liver Injury in Humans
    Downloaded from genome.cshlp.org on October 2, 2021 - Published by Cold Spring Harbor Laboratory Press Letter Mouse population-guided resequencing reveals that variants in CD44 contribute to acetaminophen-induced liver injury in humans Alison H. Harrill,1,2,12 Paul B. Watkins,3,12 Stephen Su,6 Pamela K. Ross,2 David E. Harbourt,5 Ioannis M. Stylianou,7 Gary A. Boorman,8 Mark W. Russo,3 Richard S. Sackler,9 Stephen C. Harris,11 Philip C. Smith,5 Raymond Tennant,8 Molly Bogue,7 Kenneth Paigen,7 Christopher Harris,9,10 Tanupriya Contractor,9 Timothy Wiltshire,5 Ivan Rusyn,1,2,14 and David W. Threadgill1,4,13,14,15 1Curriculum in Toxicology, University of North Carolina, Chapel Hill, North Carolina 27599, USA; 2Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, North Carolina 27599, USA; 3Division of Gastroenterology and Hepatology, University of North Carolina, Chapel Hill, North Carolina 27599, USA; 4Department of Genetics, University of North Carolina, Chapel Hill, North Carolina 27599, USA; 5School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599, USA; 6Department of Mouse Genetics, Genomics Institute of the Novartis Research Foundation, San Diego, California 92121, USA; 7The Jackson Laboratory, Bar Harbor, Maine 04609, USA; 8National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA; 9Verto Institute Research Laboratories, New Brunswick, New Jersey 08903, USA; 10Cancer Institute of New Jersey, New Brunswick, New Jersey 08903, USA; 11Purdue Pharma L.P., Stamford, Connecticut 06901, USA; 12Hamner-UNC Center for Drug Safety Sciences, The Hamner Institutes for Health Sciences, Research Triangle Park, North Carolina 27709, USA; 13Department of Genetics, North Carolina State University, Raleigh, North Carolina 27695, USA Interindividual variability in response to chemicals and drugs is a common regulatory concern.
    [Show full text]
  • PROX1 Is Associated with Cancer Progression and Prognosis in Gastric Cancer KOJI UETA 1,2 , YASUNORI OTOWA 1,2 , YOSHIHIRO KAKEJI 2 and MASANORI HIRASHIMA 1
    ANTICANCER RESEARCH 38 : 6139-6145 (2018) doi:10.21873/anticanres.12966 PROX1 Is Associated with Cancer Progression and Prognosis in Gastric Cancer KOJI UETA 1,2 , YASUNORI OTOWA 1,2 , YOSHIHIRO KAKEJI 2 and MASANORI HIRASHIMA 1 1Division of Vascular Biology, Department of Physiology and Cell Biology, and 2Division of Gastro-intestinal Surgery, Department of Surgery, Kobe University Graduate School of Medicine, Kobe, Japan Abstract. Background: It was recently reported that expected to improve prognosis. On the other hand, in the expression of prospero homeobox protein-1 (PROX1) is field of gastric cancer, trastuzumab for human epidermal correlated with the prognosis of esophageal cancer and growth factor receptor 2 (HER2)-positive gastric cancer is colorectal cancer. However, its correlation with gastric cancer the molecular-targeted therapeutic agent that has proven is unclear. Materials and Methods: Our study analyzed the useful, to date. Ramucirumab, which is an antibody against effect of PROX1 knockdown on the migration, invasion and VEGF receptor 2 has become an agent for second-line proliferation of the MKN45 human gastric cancer cell line. chemotherapy and is expected to have a therapeutic effect on The correlation between PROX1 expression levels and gastric cancer. However, options for second-line or third-line clinicopathological factors were also analyzed in tumor therapy are fewer than those for colon cancer. samples from 99 patients with gastric cancer. Results: Prospero homeobox protein-1 (PROX1) is a transcription Migration, invasion and proliferation were significantly regulator which has been implicated in differentiation of reduced in MKN45 cells with PROX1 knockdown. PROX1 lymphatic endothelial cells (1).
    [Show full text]
  • Pdpn Podoplanin
    1 Running title: Podoplanin and EMT NEW INSIGHTS INTO THE ROLE OF PODOPLANIN IN THE EPITHELIAL TO MESENCHYMAL TRANSITION Jaime Renart*, Patricia Carrasco-Ramírez, Beatriz Fernández-Muñoz1, Ester Martín- Villar, Lucía Montero, María M. Yurrita and Miguel Quintanilla. Instituto de Investigaciones Biomédicas Alberto Sols, CSIC-UAM *Corresponding author at: Instituto de Investigaciones Biomédicas Alberto Sols, CSIC- UAM. Arturo Duperier 4. 28029-Madrid. Spain. Tel: +34915854439. Fax: +34915854401. E-mail address: [email protected] 1Present address: Laboratorio Andaluz de Reprogramación Celular, Parque Tecnológico y Científico Cartuja 93. 41092-Sevilla, Spain. 2 CONTENTS 1. Introduction 2. Podoplanin 2.1. Protein structure 2.1.1. The ectodomain 2.1.2. The transmembrane domain 2.1.3. The cytoplasmic domain 2.2. Gene structure 2.3. Expression of podoplanin 2.3.1. Tissue distribution 2.3.2. Transcriptional regulation 2.3.3. Post-translational regulation 2.3.3.1. miRNAs 2.3.3.2. Glycosylation 2.3.3.3. Proteolytic processing 2.3.3.4. Phosphorylation 2.4. Podoplanin partners 2.4.1. CLEC-2 2.4.2. Tetraspanin CD9 2.4.3. Galectin 8 2.4.4. Heat shock protein A9 2.4.5. CD44 2.4.6. ERM proteins 3 2.4.7. Others 2.5. Signaling and molecular mechanisms 2.6. Podoplanin in development 2.7. Podoplanin in cancer 2.7.1. Expression in tumors 2.7.2. Role in migration, invasion and progression 2.7.3. Presence in tumor stroma 3. Epithelial to mesenchymal transition 3.1. Molecular mechanisms of EMT 3.1.1. Properties of the core Transcription factors 3.1.2.
    [Show full text]
  • Cardiac SARS‐Cov‐2 Infection Is Associated with Distinct Tran‐ Scriptomic Changes Within the Heart
    Cardiac SARS‐CoV‐2 infection is associated with distinct tran‐ scriptomic changes within the heart Diana Lindner, PhD*1,2, Hanna Bräuninger, MS*1,2, Bastian Stoffers, MS1,2, Antonia Fitzek, MD3, Kira Meißner3, Ganna Aleshcheva, PhD4, Michaela Schweizer, PhD5, Jessica Weimann, MS1, Björn Rotter, PhD9, Svenja Warnke, BSc1, Carolin Edler, MD3, Fabian Braun, MD8, Kevin Roedl, MD10, Katharina Scher‐ schel, PhD1,12,13, Felicitas Escher, MD4,6,7, Stefan Kluge, MD10, Tobias B. Huber, MD8, Benjamin Ondruschka, MD3, Heinz‐Peter‐Schultheiss, MD4, Paulus Kirchhof, MD1,2,11, Stefan Blankenberg, MD1,2, Klaus Püschel, MD3, Dirk Westermann, MD1,2 1 Department of Cardiology, University Heart and Vascular Center Hamburg, Germany. 2 DZHK (German Center for Cardiovascular Research), partner site Hamburg/Kiel/Lübeck. 3 Institute of Legal Medicine, University Medical Center Hamburg‐Eppendorf, Germany. 4 Institute for Cardiac Diagnostics and Therapy, Berlin, Germany. 5 Department of Electron Microscopy, Center for Molecular Neurobiology, University Medical Center Hamburg‐Eppendorf, Germany. 6 Department of Cardiology, Charité‐Universitaetsmedizin, Berlin, Germany. 7 DZHK (German Centre for Cardiovascular Research), partner site Berlin, Germany. 8 III. Department of Medicine, University Medical Center Hamburg‐Eppendorf, Germany. 9 GenXPro GmbH, Frankfurter Innovationszentrum, Biotechnologie (FIZ), Frankfurt am Main, Germany. 10 Department of Intensive Care Medicine, University Medical Center Hamburg‐Eppendorf, Germany. 11 Institute of Cardiovascular Sciences,
    [Show full text]
  • The Impact of Transcription Factor Prospero Homeobox 1 on the Regulation of Thyroid Cancer Malignancy
    International Journal of Molecular Sciences Review The Impact of Transcription Factor Prospero Homeobox 1 on the Regulation of Thyroid Cancer Malignancy Magdalena Rudzi ´nska 1,2 and Barbara Czarnocka 1,* 1 Department of Biochemistry and Molecular Biology, Centre of Postgraduate Medical Education, 01-813 Warsaw, Poland; [email protected] 2 Institute of Molecular Medicine, Sechenov First Moscow State Medical University, 119991 Moscow, Russia * Correspondence: [email protected]; Tel.: +48-225693812; Fax: +48-225693712 Received: 7 April 2020; Accepted: 30 April 2020; Published: 2 May 2020 Abstract: Transcription factor Prospero homeobox 1 (PROX1) is continuously expressed in the lymphatic endothelial cells, playing an essential role in their differentiation. Many reports have shown that PROX1 is implicated in cancer development and acts as an oncoprotein or suppressor in a tissue-dependent manner. Additionally, the PROX1 expression in many types of tumors has prognostic significance and is associated with patient outcomes. In our previous experimental studies, we showed that PROX1 is present in the thyroid cancer (THC) cells of different origins and has a high impact on follicular thyroid cancer (FTC) phenotypes, regulating migration, invasion, focal adhesion, cytoskeleton reorganization, and angiogenesis. Herein, we discuss the PROX1 transcript and protein structures, the expression pattern of PROX1 in THC specimens, and its epigenetic regulation. Next, we emphasize the biological processes and genes regulated by PROX1 in CGTH-W-1 cells, derived from squamous cell carcinoma of the thyroid gland. Finally, we discuss the interaction of PROX1 with other lymphatic factors. In our review, we aimed to highlight the importance of vascular molecules in cancer development and provide an update on the functionality of PROX1 in THC biology regulation.
    [Show full text]
  • The Homeobox Protein Prox1 Is a Negative Modulator of Erra/PGC
    RESEARCH COMMUNICATION metabolic genes at the transcriptional level via the action The homeobox protein Prox1 of diverse classes of transcription factors and coregulatory is a negative modulator of proteins (Desvergne et al. 2006; Feige and Auwerx 2007). Among those factors, the orphan nuclear receptor estrogen- ERRa/PGC-1a bioenergetic related receptor a (ERRa, NR3B1) and the coregulator peroxisome proliferator-activated receptor g coactivator- functions 1a (PGC-1a) have been shown to play a predominant role Alexis Charest-Marcotte,1,2 Catherine R. Dufour,1 in controlling several aspects of energy metabolism, most 1 1,2 notably mitochondrial biogenesis and oxidative phos- Brian J. Wilson, Annie M. Tremblay, ` 1,2 3,4,5 phorylation (Oxphos) (Lin et al. 2005; Giguere 2008). Lillian J. Eichner, Daniel H. Arlow, Prospero-related homeobox 1 (Prox1) is a transcription Vamsi K. Mootha,3,4,5 factor essential for the development of numerous tissues, and Vincent Gigue`re1,2,6,7,8 including the liver (Sosa-Pineda et al. 2000; Burke and Oliver 2002; Dudas et al. 2006). In particular, Prox1 plays 1Goodman Cancer Centre, McGill University, Montre´al, a critical role in determining the fate of lymphatic Que´bec H3A 1A3, Canada; 2Department of Biochemistry, endothelial cells and, consequently, Prox1-null embryos McGill University, Montre´al, Que´bec H3G 1Y6, Canada; are devoid of lymphatic vasculature and die in utero at 3Department of Systems Biology, Massachusetts General approximately embryonic day 14.5 (Wigle and Oliver Hospital, Cambridge, Massachusetts 02142, USA; 4Centre for 1999; Johnson et al. 2008). Prox1 haploinsufficient mice Human Genetic Research, Massachusetts General Hospital, also display lymphatic vascular defects that have been Cambridge, Massachusetts 02142, USA; 5Broad Institute of proposed to lead to adult-onset obesity via the promotion Massachusetts Institute of Technology/Harvard, Cambridge, of adipogenesis and increased fat storage in lymphatic- Massachusetts 02142, USA; 6Department of Medicine, McGill rich regions (Harvey et al.
    [Show full text]
  • Supplemental Data 5-21-18
    Supplemental Methods and Data Androgen receptor polyglutamine expansion drives age-dependent quality control defects and muscle dysfunction Samir R. Nath1,2,3, Zhigang Yu1, Theresa A. Gipson4, Gregory B. Marsh5, Eriko Yoshidome1, Diane M. Robins6, Sokol V. Todi5, David E. Housman4, Andrew P. Lieberman1 1 Department of Pathology, University of Michigan Medical School, Ann Arbor, MI 48109 2 Medical Scientist Training Program, University of Michigan Medical School, Ann Arbor, MI 48109 3 Cellular and Molecular Biology Graduate Program, University of Michigan Medical School, Ann Arbor, MI 48109 4 Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA 02139 5 Department of Pharmacology, Wayne State University School of Medicine, Detroit, MI 48201 6 Department of Human Genetics, University of Michigan Medical School, Ann Arbor, MI 48109 Supplemental Methods qPCR For Drosophila samples, total RNA was extracted from adult fly heads using TRIzol. Fifteen heads were used per sample. Extracted RNA was treated with TURBO DNAse (Ambion) to eliminate contaminating DNA, and reverse transcription was carried out as indicated above. RNA levels were quantified using StepOnePlus Real-Time PCR System with Fast SYBR Green Master Mix (Applied Biosystems). rp49 was used as the internal control. Each round of qRT-PCR was conducted in technical triplicates. A total of three independent repeats was conducted. Fly histology: For histological preparation (75, 76), wings and proboscises of adult flies were removed and bodies were fixed overnight in 2% glutaraldehyde/2% paraformaldehyde in Tris- buffered saline with 0.1% Triton X-100, rotating at 4˚C. Fixed bodies were subsequently dehydrated by using a series of 30%, 50%, 75%, and 100% ethanol/propylene oxide.
    [Show full text]
  • The Cataract-Linked RNA-Binding Protein Celf1 Post-Transcriptionally
    The cataract-linked RNA-binding protein Celf1 post-transcriptionally controls the spatiotemporal expression of the key homeodomain transcription factors Pax6 and Prox1 in lens development Sandeep Aryal, Justine Viet, Bailey a T Weatherbee, Archana D. Siddam, Francisco José García-Hernández, Carole Gautier-Courteille, Luc Paillard, Salil A. Lachke To cite this version: Sandeep Aryal, Justine Viet, Bailey a T Weatherbee, Archana D. Siddam, Francisco José García- Hernández, et al.. The cataract-linked RNA-binding protein Celf1 post-transcriptionally controls the spatiotemporal expression of the key homeodomain transcription factors Pax6 and Prox1 in lens development. Human Genetics, Springer Verlag, 2020, 139 (12), pp.1541-1554. 10.1007/s00439-020- 02195-7. hal-02888615 HAL Id: hal-02888615 https://hal.archives-ouvertes.fr/hal-02888615 Submitted on 10 Jul 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. The cataract-linked RNA-binding protein Celf1 post-transcriptionally controls the spatiotemporal expression of the key homeodomain transcription
    [Show full text]