DOCSLIB.ORG

Targeting Glioma Cells by Antineoplastic Activity of Reversine

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Targeting Glioma Cells by Antineoplastic Activity of Reversine ONCOLOGY LETTERS 22: 610, 2021 Targeting glioma cells by antineoplastic activity of reversine CAMILA HIRAKATA1*, KELI LIMA1*, BRUNA OLIVEIRA DE ALMEIDA1, LÍVIA BASSANI LINS DE MIRANDA1, KATHARINE GURGEL DIAS FLORÊNCIO2, LUCIANA COSTA FURTADO1, LETICIA VERAS COSTA‑LOTUFO1 and JOÃO AGOSTINHO MACHADO‑NETO1 1Department of Pharmacology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, CEP 05508‑900; 2Department of Physiology and Pharmacology, Drug Research and Development Center, Federal University of Ceará, Fortaleza, Ceará, CEP 60440‑900, Brazil Received April 3, 2021; Accepted June 2, 2021 DOI: 10.3892/ol.2021.12871 Abstract. Gliomas are the most common type of primary DNA damage and may predict improved antiglioma responses central nervous system tumors and despite great advances to the drug. Reversine may be a potential novel drug in the in understanding the molecular basis of the disease very few antineoplastic arsenal against gliomas. new therapies have been developed. Reversine, a synthetic purine analog, is a multikinase inhibitor that targets aurora Introduction kinase A (AURKA) and aurora kinase B (AURKB). In gliomas, a high expression of AURKA or AURKB is associated with Gliomas are the most common primary central nervous a malignant phenotype and a poor prognosis. The present system (CNS) tumors, corresponding to ~80% of all malig‑ study investigated reversine‑related cellular and molecular nant primary brain tumors in adults (United States of antiglioma effects in HOG, T98G and U251MG cell lines. America, 2013‑2017) (1). According to the 2016 World Health Gene and protein expression were assessed by reverse tran‑ Organization (WHO) classification of CNS tumors, diffuse scription‑quantitative PCR and western blotting, respectively. gliomas are classified based on histological (astrocytoma, For functional assays, human glioma cell lines (HOG, T98G oligoastrocytoma, oligodendroglioma and glioblastoma) and U251MG) were exposed to increasing concentrations of and genetic features [isocitrate dehydrogenase mutations, reversine (0.4‑50 µM) and subjected to various cellular and alpha‑thalassemia/mental retardation, X‑linked (ATRX) molecular assays. Reversine reduced the viability and clonoge‑ loss, TP53 mutation and 1p/19q‑codeletion] (2). In addition, nicity in a dose‑ and/or time‑dependent manner in all glioma the WHO classified CNS tumors into 4 grades (I‑IV) that cells, with HOG (high AURKB‑expression) and T98G (high have distinct prognosis and require different therapeutic AURKA‑expression) cells being more sensitive compared decisions (3,4). Glioblastomas (grade IV), which correspond with U251MG cells (low AURKA‑ and AURKB‑expression). to ~50% of gliomas are highly malignant gliomas with a Notably, HOG cells presented higher levels of polyploidy, poor prognosis (5,6). The current therapeutic options for while T98G presented multiple mitotic spindles, which is these tumors include surgery, radiation, immunotherapy and consistent with the main regulatory functions of AURKB and chemotherapy with non‑specific and highly toxic agents (7‑9). AURKA, respectively. In molecular assays, reversine reduced Currently, alkylating agents (temozolomide, carmustine and AURKA and/or AURKB expression/activity and increased lomustine), mTOR inhibitors (everolimus) and monoclonal DNA damage and apoptosis markers, but autophagy‑related antibodies (bevacizumab and naxitamab‑gqgk) are approved proteins were not modulated. In conclusion, reversine potently by the Food and Drug Administration authority for the treat‑ induced mitotic catastrophe and apoptosis in glioma cells and ment of high‑grade gliomas (https://www.cancer.gov) (10). higher basal levels of aurora kinases and genes responsive to Inspite of considerable advances in the understanding of the molecular basis of gliomas (6,11), progress in improving the clinical outcomes of the disease is still slow and very few new therapies have been developed. Reversine [2‑(4‑morpholinoanilino)‑6‑cyclohexylamino‑ Correspondence to: Professor João Agostinho Machado‑Neto, purine] is an adenosine triphosphate analog that acts as a Department of Pharmacology, Institute of Biomedical Sciences, potent multi‑kinase inhibitor selective to aurora kinases, MPS1 University of São Paulo, 1524 Avenue Professor Lineu Prestes, (monopolar spindle 1) and JNK (12‑14). The antineoplastic São Paulo, CEP 05508‑900, Brazil E‑mail: [email protected] activity of reversine has been demonstrated in hematological and solid tumors, in which it triggered cell cycle arrest, poly‑ *Contributed equally ploidy, apoptosis and autophagy (15). Aurora kinases, a family of serine/threonine kinases, Key words: gliomas, glioblastoma, aurora kinases, apoptosis, consisting of aurora kinase A (AURKA), aurora kinase B reversine (AURKB) and aurora kinase C (AURKC) serve an essential role in cell cycle progression and cellular division (16). High 2 HIRAKATA et al: ANTINEOPLASTIC EFFECTS OF REVERSINE IN GLIOMAS expression of AURKA, AURKB and AURKC has observed Inc.) for the expression of aurora kinase A (AURKA), aurora in several types of cancer, including gliomas and is associ‑ kinase B (AURKB), BCL2 apoptosis regulator (BCL2), BCL2 ated with a poor prognosis (17‑19). AURKA and AURKB are like (BCL2L1 1), baculoviral IAP repeat containing 5 (BIRC5), members with the most detailed characterizations: AURKA BCL2 interacting protein 3 (BNIP3), BCL2 interacting serves a role in the formation of the mitotic spindle during protein 3 like (BNIP3L), BCL2 associated agonist of cell death mitosis, while AURKB serves a key role in the organization of (BAD), BCL2 associated X, apoptosis regulator (BAX), BCL2 the centromere‑kinetochore region, microtubule‑kinetochore binding component 3 (BBC3), phorbol‑12‑myristate‑13‑ace‑ attachments and cytokinesis (16,20,21). Due to the differential tate‑induced protein 1 (PMAIP1), cyclin dependent kinase expression and biological functions related to the malignant inhibitor 1A (CDKN1A), cyclin dependent kinase inhibitor 1B phenotype attributed to these proteins, aurora kinases have (CDKN1B) and growth arrest and DNA damage inducible been proposed as potential targets for pharmacological inter‑ alpha (GADD45A) genes (Table SI) according to the manu‑ vention, particularly in cancer (22‑24). facturers' protocol. The thermocycling conditions included an In gliomas, high AURKA and AURKB expression is initial denaturation at 95˚C for 10 min, followed by 40 cycles associated with a poor prognosis and chemoresistance (18,25), of denaturation (95˚C for 15 sec), annealing (60˚C for 30 sec) which led to the study of the reversine effects in these disease and extension (60˚C for 30 sec). Hypoxanthine phosphoribo‑ models. Currently, reversine is in the preclinical phase of syltransferase 1 (HPRT1) and β‑actin were used as reference study in oncology, but the data accumulated thus far has genes. Relative quantification values were calculated using the indicated interesting antineoplastic activity (15). Hence, the 2‑ΔΔCt equation (26). A negative ‘No Template Control’ was identification of the most sensitive type of tumors to reversine, included for each primer pair as control for contamination including predictors of response for this drug, may identify a or unspecific amplification. Data were visualized using the profile of cancer patients eligible for clinical trials. The present multiple experiment viewer (MeV) v.4.9.0 software (27). study aimed to target aurora kinases using reversine and to identify the cellular and molecular mechanisms underlying its Western blotting. Total protein extraction was performed using anti‑glioma effects. a buffer containing 100 mM Tris (pH 7.6), 1% Triton X‑100, 150 mM NaCl, 2 mM PMSF, 10 mM Na3VO4, 100 mM NaF, Materials and methods 10 mM Na4P2O7, and 4 mM EDTA from untreated U87MG, HOG, T98G and U251MG cells, or HOG, T98G and U251MG Cell culture, reagents and chemicals. Oligodendroglioma cells treated with vehicle or reversine (1.6, 3.2 or 6.4 µM) (HOG) and glioblastoma (T98G, U251MG and U87MG) cell for 24 h or for cells treated with 1.6 µM reversine for 24, 48 lines were kindly provided by Professor Regina Pekelmann and 72 h. Extracted proteins were quantified by the Bradford Markus (University of São Paulo, São Paulo, Brazil). The method. Equal amounts of protein (30 µg) were separated by authenticity of the cells was determined by Short Tandem 8‑15% SDS‑PAGE (28) and transferred onto nitrocellulose Repeat (STR). The STR data from U87MG cells used in the membranes (GE Healthcare). Antibodies against AURKA present study is compatible with a glioblastoma of unknown (cat. no. sc‑25425), AURKB (cat. no. sc‑25426), phos‑ origin (https://web.expasy.org/cellosaurus/CVCL_0022). phorylated (p)‑histone H3S10 (sc‑8659‑R) and p‑histone H2A. All cell lines were cultured in the Roswell Park Memorial XS139 (γH2AX; sc‑51748) were purchased from Santa Cruz Institute medium (RPMI)‑1640 and supplemented with 10% Biotechnology, Inc. Antibodies against PARP1 (cat. no. 9542), fetal bovine serum (FBS), glutamine, 100 U/ml penicillin, SQSTM1/p62 (cat. no. 88588), LC3BI/II (cat. no. 2775), and 100 µg/ml streptomycin. The cells were maintained at β‑actin (cat. no. 4970) and α‑tubulin (cat. no. 2144) were 37˚C in a 5% CO2 humidified incubator and regularly split to obtained from Cell Signaling Technology, Inc. All primary maintain their exponential growth state (up to six passages). antibodies were used at 1:1,000 dilution and incubated for 16 h Reversine [2‑(4‑morpholinoanilino)‑6‑cyclohexylaminopu‑
Load more

Recommended publications
  • DNA Damage Activates a Spatially Distinct Late Cytoplasmic Cell-Cycle Checkpoint Network Controlled by MK2-Mediated RNA Stabilization
    DNA Damage Activates a Spatially Distinct Late Cytoplasmic Cell-Cycle Checkpoint Network Controlled by MK2-Mediated RNA Stabilization The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Reinhardt, H. Christian, Pia Hasskamp, Ingolf Schmedding, Sandra Morandell, Marcel A.T.M. van Vugt, XiaoZhe Wang, Rune Linding, et al. “DNA Damage Activates a Spatially Distinct Late Cytoplasmic Cell-Cycle Checkpoint Network Controlled by MK2-Mediated RNA Stabilization.” Molecular Cell 40, no. 1 (October 2010): 34–49.© 2010 Elsevier Inc. As Published http://dx.doi.org/10.1016/j.molcel.2010.09.018 Publisher Elsevier B.V. Version Final published version Citable link http://hdl.handle.net/1721.1/85107 Terms of Use Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. Molecular Cell Article DNA Damage Activates a Spatially Distinct Late Cytoplasmic Cell-Cycle Checkpoint Network Controlled by MK2-Mediated RNA Stabilization H. Christian Reinhardt,1,6,7,8 Pia Hasskamp,1,10,11 Ingolf Schmedding,1,10,11 Sandra Morandell,1 Marcel A.T.M. van Vugt,5 XiaoZhe Wang,9 Rune Linding,4 Shao-En Ong,2 David Weaver,9 Steven A. Carr,2 and Michael B. Yaffe1,2,3,* 1David H. Koch Institute for Integrative Cancer Research, Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02132, USA 2Broad Institute of MIT and Harvard, Cambridge, MA 02132, USA 3Center for Cell Decision Processes,
    [Show full text]
  • Evolution, Expression and Meiotic Behavior of Genes Involved in Chromosome Segregation of Monotremes
    G C A T T A C G G C A T genes Article Evolution, Expression and Meiotic Behavior of Genes Involved in Chromosome Segregation of Monotremes Filip Pajpach , Linda Shearwin-Whyatt and Frank Grützner * School of Biological Sciences, The University of Adelaide, Adelaide, SA 5005, Australia; fi[email protected] (F.P.); [email protected] (L.S.-W.) * Correspondence: [email protected] Abstract: Chromosome segregation at mitosis and meiosis is a highly dynamic and tightly regulated process that involves a large number of components. Due to the fundamental nature of chromosome segregation, many genes involved in this process are evolutionarily highly conserved, but duplica- tions and functional diversification has occurred in various lineages. In order to better understand the evolution of genes involved in chromosome segregation in mammals, we analyzed some of the key components in the basal mammalian lineage of egg-laying mammals. The chromosome passenger complex is a multiprotein complex central to chromosome segregation during both mitosis and meio- sis. It consists of survivin, borealin, inner centromere protein, and Aurora kinase B or C. We confirm the absence of Aurora kinase C in marsupials and show its absence in both platypus and echidna, which supports the current model of the evolution of Aurora kinases. High expression of AURKBC, an ancestor of AURKB and AURKC present in monotremes, suggests that this gene is performing all necessary meiotic functions in monotremes. Other genes of the chromosome passenger complex complex are present and conserved in monotremes, suggesting that their function has been preserved Citation: Pajpach, F.; in mammals.
    [Show full text]
  • Supplementary Information Material and Methods
    MCT-11-0474 BKM120: a potent and specific pan-PI3K inhibitor Supplementary Information Material and methods Chemicals The EGFR inhibitor NVP-AEE788 (Novartis), the Jak inhibitor I (Merck Calbiochem, #420099) and anisomycin (Alomone labs, # A-520) were prepared as 50 mM stock solutions in 100% DMSO. Doxorubicin (Adriablastin, Pfizer), EGF (Sigma Ref: E9644), PDGF (Sigma, Ref: P4306) and IL-4 (Sigma, Ref: I-4269) stock solutions were prepared as recommended by the manufacturer. For in vivo administration: Temodal (20 mg Temozolomide capsules, Essex Chemie AG, Luzern) was dissolved in 4 mL KZI/glucose (20/80, vol/vol); Taxotere was bought as 40 mg/mL solution (Sanofi Aventis, France), and prepared in KZI/glucose. Antibodies The primary antibodies used were as follows: anti-S473P-Akt (#9271), anti-T308P-Akt (#9276,), anti-S9P-GSK3β (#9336), anti-T389P-p70S6K (#9205), anti-YP/TP-Erk1/2 (#9101), anti-YP/TP-p38 (#9215), anti-YP/TP-JNK1/2 (#9101), anti-Y751P-PDGFR (#3161), anti- p21Cip1/Waf1 (#2946), anti-p27Kip1 (#2552) and anti-Ser15-p53 (#9284) antibodies were from Cell Signaling Technologies; anti-Akt (#05-591), anti-T32P-FKHRL1 (#06-952) and anti- PDGFR (#06-495) antibodies were from Upstate; anti-IGF-1R (#SC-713) and anti-EGFR (#SC-03) antibodies were from Santa Cruz; anti-GSK3α/β (#44610), anti-Y641P-Stat6 (#611566), anti-S1981P-ATM (#200-301), anti-T2609 DNA-PKcs (#GTX24194) and anti- 1 MCT-11-0474 BKM120: a potent and specific pan-PI3K inhibitor Y1316P-IGF-1R were from Bio-Source International, Becton-Dickinson, Rockland, GenTex and internal production, respectively. The 4G10 antibody was from Millipore (#05-321MG).
    [Show full text]
  • SERS and MD Simulation Studies of a Kinase Inhibitor Demonstrate the Emergence of a Potential Drug Discovery Tool
    SERS and MD simulation studies of a kinase inhibitor demonstrate the emergence of a potential drug discovery tool Dhanasekaran Karthigeyana,1, Soumik Siddhantab,1, Annavarapu Hari Kishorec,1, Sathya S. R. R. Perumald, Hans Ågrend, Surabhi Sudevana, Akshay V. Bhata, Karanam Balasubramanyama, Rangappa Kanchugarakoppal Subbegowdac,2, Tapas K. Kundua,2, and Chandrabhas Narayanab,2 aTranscription and Disease Laboratory, Molecular Biology and Genetics Unit, bLight Scattering Laboratory, Chemistry and Physics of Materials Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India; cDepartment of Studies in Chemistry, University of Mysore, Manasagangotri, Mysore 570006, India; and dDepartment of Theoretical Chemistry and Biology, School of Biotechnology, KTH Royal Institute of Technology, Roslagstullsbacken 15, SE-114 21 Stockholm, Sweden Edited by Michael L. Klein, Temple University, Philadelphia, PA, and approved May 30, 2014 (received for review February 18, 2014) We demonstrate the use of surface-enhanced Raman spectroscopy be screened for therapeutic applications. This paper provides a (SERS) as an excellent tool for identifying the binding site of small prelude to this development. This finding also facilitates the molecules on a therapeutically important protein. As an example, developing field of tip-enhanced Raman spectroscopy for im- we show the specific binding of the common antihypertension aging the small molecule interactions for in vitro and in vivo drug felodipine to the oncogenic Aurora A kinase protein via applications. A completely developed SERS–MD simulation hydrogen bonding interactions with Tyr-212 residue to specifically combination with adequate help from the structure of the pro- inhibit its activity. Based on SERS studies, molecular docking, tein may help converge potential small molecules for therapeutic molecular dynamics simulation, biochemical assays, and point applications and reduce the time for drug discovery.
    [Show full text]
  • Expression Profiling of KLF4
    Expression Profiling of KLF4 AJCR0000006 Supplemental Data Figure S1. Snapshot of enriched gene sets identified by GSEA in Klf4-null MEFs. Figure S2. Snapshot of enriched gene sets identified by GSEA in wild type MEFs. 98 Am J Cancer Res 2011;1(1):85-97 Table S1: Functional Annotation Clustering of Genes Up-Regulated in Klf4 -Null MEFs ILLUMINA_ID Gene Symbol Gene Name (Description) P -value Fold-Change Cell Cycle 8.00E-03 ILMN_1217331 Mcm6 MINICHROMOSOME MAINTENANCE DEFICIENT 6 40.36 ILMN_2723931 E2f6 E2F TRANSCRIPTION FACTOR 6 26.8 ILMN_2724570 Mapk12 MITOGEN-ACTIVATED PROTEIN KINASE 12 22.19 ILMN_1218470 Cdk2 CYCLIN-DEPENDENT KINASE 2 9.32 ILMN_1234909 Tipin TIMELESS INTERACTING PROTEIN 5.3 ILMN_1212692 Mapk13 SAPK/ERK/KINASE 4 4.96 ILMN_2666690 Cul7 CULLIN 7 2.23 ILMN_2681776 Mapk6 MITOGEN ACTIVATED PROTEIN KINASE 4 2.11 ILMN_2652909 Ddit3 DNA-DAMAGE INDUCIBLE TRANSCRIPT 3 2.07 ILMN_2742152 Gadd45a GROWTH ARREST AND DNA-DAMAGE-INDUCIBLE 45 ALPHA 1.92 ILMN_1212787 Pttg1 PITUITARY TUMOR-TRANSFORMING 1 1.8 ILMN_1216721 Cdk5 CYCLIN-DEPENDENT KINASE 5 1.78 ILMN_1227009 Gas2l1 GROWTH ARREST-SPECIFIC 2 LIKE 1 1.74 ILMN_2663009 Rassf5 RAS ASSOCIATION (RALGDS/AF-6) DOMAIN FAMILY 5 1.64 ILMN_1220454 Anapc13 ANAPHASE PROMOTING COMPLEX SUBUNIT 13 1.61 ILMN_1216213 Incenp INNER CENTROMERE PROTEIN 1.56 ILMN_1256301 Rcc2 REGULATOR OF CHROMOSOME CONDENSATION 2 1.53 Extracellular Matrix 5.80E-06 ILMN_2735184 Col18a1 PROCOLLAGEN, TYPE XVIII, ALPHA 1 51.5 ILMN_1223997 Crtap CARTILAGE ASSOCIATED PROTEIN 32.74 ILMN_2753809 Mmp3 MATRIX METALLOPEPTIDASE
    [Show full text]
  • Kinase Profiling Book
    Custom and Pre-Selected Kinase Prof iling to f it your Budget and Needs! As of July 1, 2021 19.8653 mm 128 196 12 Tyrosine Serine/Threonine Lipid Kinases Kinases Kinases Carna Biosciences, Inc. 2007 Carna Biosciences, Inc. Profiling Assays available from Carna Biosciences, Inc. As of July 1, 2021 Page Kinase Name Assay Platform Page Kinase Name Assay Platform 4 ABL(ABL1) MSA 21 EGFR[T790M/C797S/L858R] MSA 4 ABL(ABL1)[E255K] MSA 21 EGFR[T790M/L858R] MSA 4 ABL(ABL1)[T315I] MSA 21 EPHA1 MSA 4 ACK(TNK2) MSA 21 EPHA2 MSA 4 AKT1 MSA 21 EPHA3 MSA 5 AKT2 MSA 22 EPHA4 MSA 5 AKT3 MSA 22 EPHA5 MSA 5 ALK MSA 22 EPHA6 MSA 5 ALK[C1156Y] MSA 22 EPHA7 MSA 5 ALK[F1174L] MSA 22 EPHA8 MSA 6 ALK[G1202R] MSA 23 EPHB1 MSA 6 ALK[G1269A] MSA 23 EPHB2 MSA 6 ALK[L1196M] MSA 23 EPHB3 MSA 6 ALK[R1275Q] MSA 23 EPHB4 MSA 6 ALK[T1151_L1152insT] MSA 23 Erk1(MAPK3) MSA 7 EML4-ALK MSA 24 Erk2(MAPK1) MSA 7 NPM1-ALK MSA 24 Erk5(MAPK7) MSA 7 AMPKα1/β1/γ1(PRKAA1/B1/G1) MSA 24 FAK(PTK2) MSA 7 AMPKα2/β1/γ1(PRKAA2/B1/G1) MSA 24 FER MSA 7 ARG(ABL2) MSA 24 FES MSA 8 AurA(AURKA) MSA 25 FGFR1 MSA 8 AurA(AURKA)/TPX2 MSA 25 FGFR1[V561M] MSA 8 AurB(AURKB)/INCENP MSA 25 FGFR2 MSA 8 AurC(AURKC) MSA 25 FGFR2[V564I] MSA 8 AXL MSA 25 FGFR3 MSA 9 BLK MSA 26 FGFR3[K650E] MSA 9 BMX MSA 26 FGFR3[K650M] MSA 9 BRK(PTK6) MSA 26 FGFR3[V555L] MSA 9 BRSK1 MSA 26 FGFR3[V555M] MSA 9 BRSK2 MSA 26 FGFR4 MSA 10 BTK MSA 27 FGFR4[N535K] MSA 10 BTK[C481S] MSA 27 FGFR4[V550E] MSA 10 BUB1/BUB3 MSA 27 FGFR4[V550L] MSA 10 CaMK1α(CAMK1) MSA 27 FGR MSA 10 CaMK1δ(CAMK1D) MSA 27 FLT1 MSA 11 CaMK2α(CAMK2A) MSA 28
    [Show full text]
  • Phosphorylation of Threonine 3 on Histone H3 by Haspin Kinase Is Required for Meiosis I in Mouse Oocytes
    ß 2014. Published by The Company of Biologists Ltd | Journal of Cell Science (2014) 127, 5066–5078 doi:10.1242/jcs.158840 RESEARCH ARTICLE Phosphorylation of threonine 3 on histone H3 by haspin kinase is required for meiosis I in mouse oocytes Alexandra L. Nguyen, Amanda S. Gentilello, Ahmed Z. Balboula*, Vibha Shrivastava, Jacob Ohring and Karen Schindler` ABSTRACT a lesser extent. However, it is not known whether haspin is required for meiosis in oocytes. To date, the only known haspin substrates Meiosis I (MI), the division that generates haploids, is prone to are threonine 3 of histone H3 (H3T3), serine 137 of macroH2A and errors that lead to aneuploidy in females. Haspin is a kinase that threonine 57 of CENP-T (Maiolica et al., 2014). Knockdown or phosphorylates histone H3 on threonine 3, thereby recruiting Aurora inhibition of haspin in mitotically dividing tissue culture cell kinase B (AURKB) and the chromosomal passenger complex lines reveal that phosphorylation of H3T3 is essential for the (CPC) to kinetochores to regulate mitosis. Haspin and AURKC, an alignment of chromosomes at the metaphase plate (Dai and AURKB homolog, are enriched in germ cells, yet their significance Higgins, 2005; Dai et al., 2005), regulation of chromosome in regulating MI is not fully understood. Using inhibitors and cohesion (Dai et al., 2009) and establishing a bipolar spindle (Dai overexpression approaches, we show a role for haspin during MI et al., 2009). In mitotic metaphase, phosphorylation of H3T3 is in mouse oocytes. Haspin-perturbed oocytes display abnormalities restricted to kinetochores, and this mark signals recruitment of the in chromosome morphology and alignment, improper kinetochore– chromosomal passenger complex (CPC) (Dai et al., 2005; Wang microtubule attachments at metaphase I and aneuploidy at et al., 2010; Yamagishi et al., 2010).
    [Show full text]
  • Gadd45a Regulates B-Catenin Distribution and Maintains Cell–Cell Adhesion/ Contact
    Oncogene (2007) 26, 6396–6405 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc ORIGINAL ARTICLE Gadd45a regulates b-catenin distribution and maintains cell–cell adhesion/ contact JJi1, R Liu1, T Tong, Y Song, S Jin, M Wu and Q Zhan State Key Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China Gadd45a,a growth arrest and DNA-damage gene,plays 1999;Jin et al., 2000;Tong et al., 2005). Gadd45a important roles in the control of cell cycle checkpoints, also plays roles in negative regulation of cell malig- DNA repair and apoptosis. We show here that Gadd45a is nancy. Mouse embryonic fibroblasts (MEFs) derived involved in the control of cell contact inhibition and cell– from Gadd45a-null mice exhibited genomic instability, cell adhesion. Gadd45a can serve as an adapter to enhance single oncogene-mediated transformation, loss of nor- the interaction between b-catenin and Caveolin-1,and in mal cellular senescence, increased cellular proliferation, turn induces b-catenin translocation to cell membrane for centrosome amplification and reduced DNA repair maintaining cell–cell adhesion/contact inhibition. This is (Hollander et al., 1999;Smith et al., 2000). Gadd45a- coupled with reduction of b-catenin in cytoplasm and null mice have an increased frequency of tumorigenesis nucleus following Gadd45a induction,which is reflected by by ionizing radiation (IR), ultraviolet radiation (UVR) the downregulation of cyclin D1,one of the b-catenin and dimethylbenzanthracene (DMBA), although these targeted genes. Additionally,Gadd45a facilitates ultra- mice do not develop spontaneous tumors (Hollander violet radiation-induced degradation of cytoplasmic and et al., 1999, 2001;Hildesheim et al., 2002).
    [Show full text]
  • A New AURKC Mutation Causing Macrozoospermia
    A new AURKC mutation causing macrozoospermia: implications for human spermatogenesis and clinical diagnosis Mariem Ben Khelifa 1 2 3 , Raoudha Zouari 4 , Radu Harbuz 2 1 , Lazhar Halouani 4 , Christophe Arnoult 1 , Joël Lunardi 2 5 , Pierre F. Ray 1 2 * 1 AGIM, AGeing and IMagery, CNRS FRE3405 Université Joseph Fourier - Grenoble I , Ecole Pratique des Hautes Etudes , CNRS : UMR5525 , Faculté de médecine de Grenoble, 38700 La Tronche,FR 2 Laboratoire de biochimie et génétique moléculaire CHU Grenoble , 38043 Grenoble,FR 3 Molecular Investigation of Genetic Orphan Diseases Research Unit Institut Pasteur de Tunis , Research Unit UR04/SP03,TN 4 Clinique de la reproduction les Jasmins Clinique de la reproduction les Jasmins , 23, Av. Louis BRAILLE, 1002 Tunis,TN 5 GIN, Grenoble Institut des Neurosciences INSERM : U836 , CEA , Université Joseph Fourier - Grenoble I , CHU Grenoble , UJF - Site Santé La Tronche BP 170 38042 Grenoble Cedex 9,FR * Correspondence should be addressed to: Pierre Ray <[email protected] > Abstract The presence of close to 100% large-headed multi-tailed spermatozoa in the ejaculate has been described as a rare phenotype of male infertility with a very poor prognosis. We demonstrated previously that most cases were caused by a homozygous mutation (c.144delC) in the Aurora Kinase C gene (AURKC) leading to the absence or the production of a nonfunctional protein. AURKC deficiency in these patients blocked meiosis and resulted in the production of tetraploid spermatozoa unsuitable for fertilization. We describe here the study of two brothers presenting with large-headed spermatozoa. Molecular analysis of the AURKC gene was carried out in two brothers presenting with a typical large headed spermatozoa phenotype.
    [Show full text]
  • Aberrant Gene Expression: Diagnostic Markers and Therapeutic Targets for Pancreatic Cancer
    ABERRANT GENE EXPRESSION: DIAGNOSTIC MARKERS AND THERAPEUTIC TARGETS FOR PANCREATIC CANCER Jeran Kent Stratford A dissertation submitted to the faculty of the University of North Carolina at Chapel Hill in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Pharmacology. Chapel Hill 2014 Approved by: Jen Jen Yeh Channing J. Der Gary L. Johnson Adrienne D. Cox Timothy C. Elston © 2014 Jeran Kent Stratford ALL RIGHTS RESERVED ii ABSTRACT Jeran Kent Stratford: Aberrant gene expression: diagnostic markers and therapeutic targets for pancreatic cancer (Under the direction of Jen Jen Yeh and Channing J. Der) Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer and the fourth leading cause of cancer-related death in the United States. The overall median survival for patients diagnosed with PDAC is five to eight months. The poor outcome is due, in part, to a lack of disease-specific symptoms that can be used for early detection, and as such, most patients present with locally advanced or metastatic disease at the time of diagnosis. Therefore, the need for diagnostic tools is both great and urgent. Furthermore, current chemotherapies have low response rates and high toxicity, limiting their use, and there are currently no effective targeted therapies for PDAC. Therefore, a greater understanding of the underlying biology of pancreatic cancer is needed to identify tumor-specific vulnerabilities that can be therapeutically exploited. Pancreatic cancer development is driven by genomic changes that alter gene expression. Aberrant gene expression produces changes in protein expression, which in turn may confer growth advantages to the tumor; often the tumor then develops a dependency on continued aberrant gene and protein expression.
    [Show full text]
  • The Ins and Outs of Autophagic Ribosome Turnover
    Biochemistry, Biophysics and Molecular Biology Publications Biochemistry, Biophysics and Molecular Biology 2019 The Ins and Outs of Autophagic Ribosome Turnover Zakayo Kazibwe Iowa State University, [email protected] Ang-Yu Liu Iowa State University, [email protected] Gustavo C. Macintosh Iowa State University, [email protected] Diane C. Bassham Iowa State University, [email protected] Follow this and additional works at: https://lib.dr.iastate.edu/bbmb_ag_pubs Part of the Cell and Developmental Biology Commons, Genetics Commons, and the Molecular Biology Commons The complete bibliographic information for this item can be found at https://lib.dr.iastate.edu/ bbmb_ag_pubs/260. For information on how to cite this item, please visit http://lib.dr.iastate.edu/howtocite.html. This Article is brought to you for free and open access by the Biochemistry, Biophysics and Molecular Biology at Iowa State University Digital Repository. It has been accepted for inclusion in Biochemistry, Biophysics and Molecular Biology Publications by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. The Ins and Outs of Autophagic Ribosome Turnover Abstract Ribosomes are essential for protein synthesis in all organisms and their biogenesis and number are tightly controlled to maintain homeostasis in changing environmental conditions. While ribosome assembly and quality control mechanisms have been extensively studied, our understanding of ribosome degradation is limited. In yeast or animal cells, ribosomes are degraded after transfer into the vacuole or lysosome by ribophagy or nonselective autophagy, and ribosomal RNA can also be transferred directly across the lysosomal membrane by RNautophagy. In plants, ribosomal RNA is degraded by the vacuolar T2 ribonuclease RNS2 after transport by autophagy-related mechanisms, although it is unknown if a selective ribophagy pathway exists in plants.
    [Show full text]
  • DNA Excision Repair Proteins and Gadd45 As Molecular Players for Active DNA Demethylation
    Cell Cycle ISSN: 1538-4101 (Print) 1551-4005 (Online) Journal homepage: http://www.tandfonline.com/loi/kccy20 DNA excision repair proteins and Gadd45 as molecular players for active DNA demethylation Dengke K. Ma, Junjie U. Guo, Guo-li Ming & Hongjun Song To cite this article: Dengke K. Ma, Junjie U. Guo, Guo-li Ming & Hongjun Song (2009) DNA excision repair proteins and Gadd45 as molecular players for active DNA demethylation, Cell Cycle, 8:10, 1526-1531, DOI: 10.4161/cc.8.10.8500 To link to this article: http://dx.doi.org/10.4161/cc.8.10.8500 Published online: 15 May 2009. Submit your article to this journal Article views: 135 View related articles Citing articles: 92 View citing articles Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=kccy20 Download by: [University of Pennsylvania] Date: 27 April 2017, At: 12:48 [Cell Cycle 8:10, 1526-1531; 15 May 2009]; ©2009 Landes Bioscience Perspective DNA excision repair proteins and Gadd45 as molecular players for active DNA demethylation Dengke K. Ma,1,2,* Junjie U. Guo,1,3 Guo-li Ming1-3 and Hongjun Song1-3 1Institute for Cell Engineering; 2Department of Neurology; and 3The Solomon Snyder Department of Neuroscience; Johns Hopkins University School of Medicine; Baltimore, MD USA Abbreviations: DNMT, DNA methyltransferases; PGCs, primordial germ cells; MBD, methyl-CpG binding protein; NER, nucleotide excision repair; BER, base excision repair; AP, apurinic/apyrimidinic; SAM, S-adenosyl methionine Key words: DNA demethylation, Gadd45, Gadd45a, Gadd45b, Gadd45g, 5-methylcytosine, deaminase, glycosylase, base excision repair, nucleotide excision repair DNA cytosine methylation represents an intrinsic modifica- silencing of gene activity or parasitic genetic elements (Fig.
    [Show full text]