DOCSLIB.ORG

Mucin Glycosylating Enzyme GALNT2 Regulates the Malignant Character of Hepatocellular Carcinoma by Modifying the EGF Receptor

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mucin Glycosylating Enzyme GALNT2 Regulates the Malignant Character of Hepatocellular Carcinoma by Modifying the EGF Receptor Published OnlineFirst October 11, 2011; DOI: 10.1158/0008-5472.CAN-11-1161 Cancer Tumor and Stem Cell Biology Research Mucin Glycosylating Enzyme GALNT2 Regulates the Malignant Character of Hepatocellular Carcinoma by Modifying the EGF Receptor Yao-Ming Wu1,4, Chiung-Hui Liu3,4, Rey-Heng Hu1, Miao-Juei Huang3,4, Jian-Jr Lee3, Chi-Hau Chen2,3, John Huang1, Hong-Shiee Lai1, Po-Huang Lee1, Wen-Ming Hsu1,4, Hsiu-Chin Huang5, and Min-Chuan Huang3,4 Abstract Extracellular glycosylation is a critical determinant of malignant character. Here, we report that N-acetylga- lactosaminyltransferase 2 (GALNT2), the enzyme that mediates the initial step of mucin type-O glycosylation, is a critical mediator of malignant character in hepatocellular carcinoma (HCC) that acts by modifying the activity of the epidermal growth factor receptor (EGFR). GALNT2 mRNA and protein were downregulated frequently in HCC tumors where these events were associated with vascular invasion and recurrence. Restoring GALNT2 expression in HCC cells suppressed EGF-induced cell growth, migration, and invasion in vitro and in vivo. Mechanistic investigations revealed that the status of the O-glycans attached to the EGFR was altered by GALNT2, changing EGFR responses after EGF binding. Inhibiting EGFR activity with erlotinib decreased the malignant characters caused by siRNA-mediated knockdown of GALNT2 in HCC cells, establishing the critical role of EGFR in mediating the effects of GALNT2 expression. Taken together, our results suggest that GALNT2 dysregulation contributes to the malignant behavior of HCC cells, and they provide novel insights into the significance of O-glycosylation in EGFR activity and HCC pathogenesis. Cancer Res; 71(23); 7270–9. Ó2011 AACR. Introduction Tumor-associated carbohydrate antigens have drawn global attention to develop diagnostic reagents and vaccines for Hepatocellular carcinoma (HCC) is the sixth most common cancer therapy (3). However, the glycogenes responsible for malignancy and the third leading cause of cancer-related death the expression of these antigens and their pathophysiologic worldwide (1). The primary curative treatment for HCC is roles in human cancers are still largely unknown. hepatic resection. Although clinical treatment of HCC is con- Two major types of protein glycosylation in mammalian tinuously evolving, the prognosis of HCC patients remains cells exist: N-linked and O-linked. The most frequently occur- poor. To improve the survival of HCC patients, further under- ring O-glycosylation is the mucin type, initiated by the transfer standing of HCC pathogenesis and novel treatment agents are of UDP-N-acetylgalactosamine (UDP-GalNAc) to the hydroxyl needed. group of serine (S) or threonine (T) residue forming Tn antigen Glycosylation is the most common posttranslational mod- (GalNAca-S/T; ref. 4). This reaction is catalyzed by a large ification of proteins. Aberrant glycosylation is a hallmark of family of polypeptide GalNAc transferases (GALNT), consisting most human cancers and affects many cellular properties, of at least 20 members in humans, namely GALNT1 to 14 and including cell proliferation, apoptosis, differentiation, trans- GALNTL1 to L6 (5, 6). formation, migration, invasion, and immune responses (2). Studies have shown that O-glycans and GALNT genes play critical roles in a variety of biological functions and human disease development. For instance, loss of GALNT1 activity in Authors' Affiliations: Departments of 1Surgery and Obstetrics and 2 3 mice results in bleeding disorder (7). Risk of epithelial ovarian Gynecology, National Taiwan University Hospital; Graduate Institute of Anatomy and Cell Biology, National Taiwan University College of cancer (8) and coronary artery disease (9) have been associated Medicine; 4Research Center for Developmental Biology and Regener- GALNT1 5 with single nucleotide polymorphisms of and ative Medicine, National Taiwan University, Taipei; and Animal Tech- GALNT2 nology Institute Taiwan, Miaoli, Taiwan , respectively. GALNT3 expression is a potential diag- nostic marker for lung (10) and pancreatic (11) cancers. Note: Supplementary data for this article are available at Cancer Research fi Online (http://cancerres.aacrjournals.org/). GALNT6 modi es mucin 1 glycosylation and regulates prolif- eration of breast cancer cells (12). Corresponding Author: Min-Chuan Huang, Graduate Institute of Anatomy and Cell Biology, National Taiwan University College of Medicine, No. 1, The epidermal growth factor receptor (EGFR) is a promising Sec. 1 Jen-Ai Road, Taipei 100, Taiwan. Phone: 886-2-23123456 (ext. therapeutic target as its overexpression is associated with 88177); Fax: 886-2-23915292; E-mail: [email protected] various cancers and plays a crucial role in tumor malignancy doi: 10.1158/0008-5472.CAN-11-1161 (13). Overexpression of EGFR in HCC (14) and upregulation of Ó2011 American Association for Cancer Research. EGF in advanced HCC compared with the control liver tissue 7270 Cancer Res; 71(23) December 1, 2011 Downloaded from cancerres.aacrjournals.org on October 3, 2021. © 2011 American Association for Cancer Research. Published OnlineFirst October 11, 2011; DOI: 10.1158/0008-5472.CAN-11-1161 GALNT2 in Hepatocellular Carcinoma and early HCC (15) suggest the potential role of EGFR-ligand Immunohistochemistry interaction in HCC progression. Phase 2 clinical trials of Paraffin-embedded tissue sections were incubated with erlotinib, an EGFR inhibitor, showed 9% partial response and anti-GALNT2 polyclonal antibody (1:75, Sigma) diluted with 25% partial response combined with bevacizumab (antibody 5% nonfat milk/PBS for 16 hours at 4C. After rinsing twice for VEGF) for advanced HCC (16). The phase 3 clinical trial of with PBS, Super Sensitive Link-Label immunohistochemistry erlotinib for advanced HCC is still ongoing. However, the Detection System (BioGenex) was used and the specific immu- clinical efficacy is still unsatisfactory. Thus, to improve the nostaining was visualized with 3,3-diaminobenzidine liquid effect of EGFR-targeted therapies, molecular mechanisms by substrate system (Sigma). All sections were counterstained which EGFR regulates HCC properties should be further with hematoxylin. Negative controls were done by replacing investigated. primary antibody with control IgG. Tumor cell proliferation In HCC, the expression pattern and function of GALNT was assessed by Ki67 immunoreactivity. Anti-Ki67 rabbit family have never been reported, although O-glycosylation can polyclonal antibody (Vector Laboratories) was applied to the regulate multiple cellular properties. Here, we report that slides at 1:500 dilution. Cells with positively stained nuclei were GALNT2 is frequently downregulated in HCC. Moreover, counted in 5 random fields. GALNT2 modifies EGFR O-glycosylation and activity, and plays a critical role in the malignant phenotype of HCC cells in vitro Plasmid construction and in vivo. Reverse transcriptase PCR (RT-PCR) was done for cloning of full-length human GALNT2 (Accession No. NM_004481) from Materials and Methods nontumorous liver total RNA (BD Biosciences). The sense primer was 50-ATGCGGCGGCGCTCGCGGAT-30, and the anti- Tissue samples sense primer was 50-CTGCTGCAGGTTGAGCGTGA-30. The Postsurgery fresh tissue samples were collected from RT-PCR products were cloned into pcDNA3.1/myc-His (Invi- patients receiving treatment at the National Taiwan University trogen Life Technologies) to generate the GALNT2/myc-His Hospital (Supplementary Table S1). The tumor samples were fusion gene. The insert was confirmed by DNA sequencing. taken from the central part of the resected tumor and the paired nontumor samples were taken 2 cm away from the Transfection tumor. For immunohistochemistry, specimens were fixed in Overexpression of GALNT2 gene was achieved by transfect- 4% (w/v) paraformaldehyde/PBS. For RNA extraction, speci- ing cells with pcDNA3.1/GALNT2/mycHis plasmids using Lipo- mens were soaked in RNAlater (Qiagen Corp.) at 4C overnight fectamine 2000 (Invitrogen, Life Technologies) according to and then stored at À20C. Samples used for Western blotting the manufacturer's protocol. The transfected cells were select- were stored at À80C. Ethics approval was obtained from the ed with 600 mg/mL of G418 for 14 days and then pooled for local hospital ethic committees and a written consent was further studies. obtained from each patient before sample collection. siRNA knockdown of GALNT2 expression Cell line and cell culture SMARTpool siRNA oligonucleotides against GALNT2 and Human liver cancer cell lines Huh7, PLC5, and HepG2 siCONTROL nontargeting siRNA were synthesized by Dhar- were purchased from Bioresource Collection and Research macon Research (Thermoscientific). For knockdown of Center (Hsinchu, Taiwan) in 2008. HA22T, SUN387, and GALNT2, cells were transfected with siRNA using Dharma- HCC36 cells were kindly provided by Shiou-Hwei Yeh FECT 4 (Thermoscientific) with a final concentration of 100 (National Taiwan University, Taiwan) in 2010. All cell lines nmol siRNA for 48 hours. were authenticated by the provider based on morphology, antigen expression, growth, DNA profile, and cytogenetics. Western blot analysis Cells were maintained with Dulbecco's modified Eagle's GALNT2 proteins were detected with rabbit anti-GALNT2 medium (DMEM; Biowest) containing 10% FBS (PAA Lab- polyclonal antibody (Sigma). For detection of EGFR and its oratories), 100 IU/mL penicillin, and 100 mg/mL streptomy- downstream signaling molecules, anti-phospho-tyrosine anti- cin (Biowest) in tissue culture incubator
Load more

Recommended publications
  • Co-Occupancy by Multiple Cardiac Transcription Factors Identifies
    Co-occupancy by multiple cardiac transcription factors identifies transcriptional enhancers active in heart Aibin Hea,b,1, Sek Won Konga,b,c,1, Qing Maa,b, and William T. Pua,b,2 aDepartment of Cardiology and cChildren’s Hospital Informatics Program, Children’s Hospital Boston, Boston, MA 02115; and bHarvard Stem Cell Institute, Harvard University, Cambridge, MA 02138 Edited by Eric N. Olson, University of Texas Southwestern, Dallas, TX, and approved February 23, 2011 (received for review November 12, 2010) Identification of genomic regions that control tissue-specific gene study of a handful of model genes (e.g., refs. 7–10), it has not been expression is currently problematic. ChIP and high-throughput se- evaluated using unbiased, genome-wide approaches. quencing (ChIP-seq) of enhancer-associated proteins such as p300 In this study, we used a modified ChIP-seq approach to define identifies some but not all enhancers active in a tissue. Here we genome wide the binding sites of these cardiac TFs (1). We show that co-occupancy of a chromatin region by multiple tran- provide unbiased support for collaborative TF interactions in scription factors (TFs) identifies a distinct set of enhancers. GATA- driving cardiac gene expression and use this principle to show that chromatin co-occupancy by multiple TFs identifies enhancers binding protein 4 (GATA4), NK2 transcription factor-related, lo- with cardiac activity in vivo. The majority of these multiple TF- cus 5 (NKX2-5), T-box 5 (TBX5), serum response factor (SRF), and “ binding loci (MTL) enhancers were distinct from p300-bound myocyte-enhancer factor 2A (MEF2A), here referred to as cardiac enhancers in location and functional properties.
    [Show full text]
  • Human Lectins, Their Carbohydrate Affinities and Where to Find Them
    biomolecules Review Human Lectins, Their Carbohydrate Affinities and Where to Review HumanFind Them Lectins, Their Carbohydrate Affinities and Where to FindCláudia ThemD. Raposo 1,*, André B. Canelas 2 and M. Teresa Barros 1 1, 2 1 Cláudia D. Raposo * , Andr1 é LAQVB. Canelas‐Requimte,and Department M. Teresa of Chemistry, Barros NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829‐516 Caparica, Portugal; [email protected] 12 GlanbiaLAQV-Requimte,‐AgriChemWhey, Department Lisheen of Chemistry, Mine, Killoran, NOVA Moyne, School E41 of ScienceR622 Co. and Tipperary, Technology, Ireland; canelas‐ [email protected] NOVA de Lisboa, 2829-516 Caparica, Portugal; [email protected] 2* Correspondence:Glanbia-AgriChemWhey, [email protected]; Lisheen Mine, Tel.: Killoran, +351‐212948550 Moyne, E41 R622 Tipperary, Ireland; [email protected] * Correspondence: [email protected]; Tel.: +351-212948550 Abstract: Lectins are a class of proteins responsible for several biological roles such as cell‐cell in‐ Abstract:teractions,Lectins signaling are pathways, a class of and proteins several responsible innate immune for several responses biological against roles pathogens. such as Since cell-cell lec‐ interactions,tins are able signalingto bind to pathways, carbohydrates, and several they can innate be a immuneviable target responses for targeted against drug pathogens. delivery Since sys‐ lectinstems. In are fact, able several to bind lectins to carbohydrates, were approved they by canFood be and a viable Drug targetAdministration for targeted for drugthat purpose. delivery systems.Information In fact, about several specific lectins carbohydrate were approved recognition by Food by andlectin Drug receptors Administration was gathered for that herein, purpose. plus Informationthe specific organs about specific where those carbohydrate lectins can recognition be found by within lectin the receptors human was body.
    [Show full text]
  • Gene Expression Responses to DNA Damage Are Altered in Human Aging and in Werner Syndrome
    Oncogene (2005) 24, 5026–5042 & 2005 Nature Publishing Group All rights reserved 0950-9232/05 $30.00 www.nature.com/onc Gene expression responses to DNA damage are altered in human aging and in Werner Syndrome Kasper J Kyng1,2, Alfred May1, Tinna Stevnsner2, Kevin G Becker3, Steen Klvra˚ 4 and Vilhelm A Bohr*,1 1Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, 5600 Nathan Shock Drive, Baltimore, MD 21224, USA; 2Danish Center for Molecular Gerontology, Department of Molecular Biology, University of Aarhus, DK-8000 Aarhus C, Denmark; 3Gene Expression and Genomics Unit, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA; 4Institute for Human Genetics, University of Aarhus, Denmark The accumulation of DNA damage and mutations is syndromes, caused by heritable mutations inactivating considered a major cause of cancer and aging. While it is proteins that sense or repair DNA damage, which known that DNA damage can affect changes in gene accelerate some but not all signs of normal aging (Hasty expression, transcriptional regulation after DNA damage et al., 2003). Age is associated withan increase in is poorly understood. We characterized the expression of susceptibility to various forms of stress, and sporadic 6912 genes in human primary fibroblasts after exposure to reports suggest that an age-related decrease in DNA three different kinds of cellular stress that introduces repair may increase the susceptibility of cells to agents DNA damage: 4-nitroquinoline-1-oxide (4NQO), c-irra- causing DNA damage. Reduced base excision repair has diation, or UV-irradiation. Each type of stress elicited been demonstrated in nuclear extracts from aged human damage specific gene expression changes of up to 10-fold.
    [Show full text]
  • Investigation of Adiposity Phenotypes in AA Associated with GALNT10 & Related Pathway Genes
    Investigation of Adiposity Phenotypes in AA Associated With GALNT10 & Related Pathway Genes By Mary E. Stromberg A Dissertation Submitted to the Graduate Faculty of WAKE FOREST UNIVERSITY GRADUATE SCHOOL OF ARTS AND SCIENCES in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY In Molecular Genetics and Genomics December 2018 Winston-Salem, North Carolina Approved by: Donald W. Bowden, Ph.D., Advisor Maggie C.Y. Ng, Ph.D., Advisor Timothy D. Howard, Ph.D., Chair Swapan Das, Ph.D. John P. Parks, Ph.D. Acknowledgements I would first like to thank my mentors, Dr. Bowden and Dr. Ng, for guiding my learning and growth during my years at Wake Forest University School of Medicine. Thank you Dr. Ng for spending so much time ensuring that I learn every detail of every protocol, and supporting me through personal difficulties over the years. Thank you Dr. Bowden for your guidance in making me a better scientist and person. I would like to thank my committee for their patience and the countless meetings we have had in discussing this project. I would like to say thank you to the members of our lab as well as the Parks lab for their support and friendship as well as their contributions to my project. Special thanks to Dean Godwin for his support and understanding. The umbrella program here at WFU has given me the chance to meet some of the best friends I could have wished for. I would like to also thank those who have taught me along the way and helped me to get to this point of my life, with special thanks to the late Dr.
    [Show full text]
  • Bioinformatic Analysis Reveals Novel Hub Genes Pathways Associated
    Bioinformatic Analysis Reveals Novel Hub Genes pathways associated with IgA nephropathy xue Jiang Hangzhou Hospital of Traditional Chinese Medicine Zhijie Xu Zhejiang University School of Medicine First Aliated Hospital Yuanyuan Du Hangzhou Hospital of Traditional Chinese Medicine Hongyu Chen ( [email protected] ) Hangzhou Hospital of Traditional Chinese Medicine https://orcid.org/0000-0002-8207-1421 Research Keywords: IgA nephropathy, Gene Expression Proling, Bioinformatic Analysis Posted Date: May 22nd, 2020 DOI: https://doi.org/10.21203/rs.3.rs-29974/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Version of Record: A version of this preprint was published on September 7th, 2020. See the published version at https://doi.org/10.1186/s40001-020-00441-2. Page 1/18 Abstract Background Immunoglobulin A nephropathy (IgAN) is the most common primary glomerulopathy worldwide. However, the molecular events underlying IgAN remain to be fully elucidated. The aim of the study is to identify novel biomarkers of IgAN through bioinformatics analysis and elucidate the possible molecular mechanism. Methods Based on the microarray data GSE93798 and GSE37460 were downloaded from the Gene Expression Omnibus database, the differentially expressed genes (DEGs) between IgAN samples and normal controls were identied. With DEGs, we further performed a series of functional enrichment analyses. Protein- protein interaction (PPI) networks of the DEGs were built with the STRING online search tool and visualized by using Cytoscape, then further identied the hub gene and most important module in DEGs, Biological Networks Gene Oncology tool (BiNGO) were then performed to elucidate the molecular mechanism of IgAN.
    [Show full text]
  • Recycling of Golgi Glycosyltransferases Requires Direct Binding to Coatomer
    Recycling of Golgi glycosyltransferases requires direct binding to coatomer Lin Liua, Balraj Doraya, and Stuart Kornfelda,1 aDepartment of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110 Contributed by Stuart Kornfeld, July 20, 2018 (sent for review June 14, 2018; reviewed by Juan S. Bonifacino and Suzanne R. Pfeffer) The glycosyltransferases of the mammalian Golgi complex must Results recycle between the stacked cisternae of that organelle to Specific Binding of Ptase N-Tail to Coatomer. Ptase is a type III maintain their proper steady-state localization. This trafficking transmembrane protein with a 17-amino acid N-tail and a 21- is mediated by COPI-coated vesicles, but how the glycosyltrans- amino acid C-tail, both of which are cytoplasmically oriented. ferases are incorporated into these transport vesicles is poorly Previously we showed that two mutations in the N-tail, Lys4Gln understood. Here we show that the N-terminal cytoplasmic tails (K4Q) and Ser15Tyr (S15Y), identified in patients with the ly- cis (N-tails) of a number of Golgi glycosyltransferases which sosomal storage disorder mucolipidosis III, resulted in the ϕ δ share a -(K/R)-X-L-X-(K/R) sequence bind directly to the -and transferase being mislocalized to punctae tentatively identified as ζ -subunits of COPI. Mutations of this N-tail motif impair binding the endolysosomal compartment (15). We now document that to the COPI subunits, leading to mislocalization of the transfer- these mutant proteins, along with a protein containing a third ases to lysosomes. The physiological importance of these inter- patient mutation, Arg8Gly (R8G) (16), colocalize with the ly- actions is illustrated by mucolipidosis III patients with missense sosomal marker cathepsin D, establishing that the punctae rep- mutations in the N-tail of GlcNAc-1-phosphotransferase that resent mostly lysosomes (Fig.
    [Show full text]
  • Probing Isoform-Specific Functions of Polypeptide Galnac-Transferases
    Probing isoform-specific functions of polypeptide GalNAc-transferases using zinc finger nuclease glycoengineered SimpleCells Katrine T.-B. G. Schjoldager, Sergey Y. Vakhrushev, Yun Kong, Catharina Steentoft, Aaron S. Nudelman, Nis B. Pedersen, Hans H. Wandall, Ulla Mandel, Eric P. Bennett, Steven B. Levery, and Henrik Clausen1 Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine and School of Dentistry, Faculty of Health Sciences, University of Copenhagen, DK-2200 Copenhagen N, Denmark Edited by Stuart A. Kornfeld, Washington University School of Medicine, St. Louis, MO, and approved April 10, 2012 (received for review March 1, 2012) Our knowledge of the O-glycoproteome [N-acetylgalactosamine protein function, with the most recently discovered function being (GalNAc) type] is highly limited. The O-glycoproteome is differentially coregulation of proprotein convertase processing of proteins (6, regulated in cells by dynamic expression of a subset of 20 polypeptide 9). Compared with other types of protein glycosylation, GalNAc- GalNAc-transferases (GalNAc-Ts), and methods to identify important type O-glycosylation is unique with its 20 distinct isoenzymes (5, functions of individual GalNAc-Ts are largely unavailable. We recently 8). This allows for an incomparable level of cell- and protein- specific, as well as dynamic, regulation of site-directed glycosyla- introduced SimpleCells, i.e., human cell lines made deficient in O-gly- – fi tion, which can be essential for development and health (8 10). can extension by zinc nger nuclease targeting of a key gene in O- However, technical limitations hamper our understanding of the glycan elongation (Cosmc), which allows for proteome-wide discovery O-glycoproteome and functions of site-specific O-glycosylation.
    [Show full text]
  • A Polymorphism in the GALNT2 Gene and Ovarian Cancer Risk in Four Population Based Case-Control Studies
    A polymorphism in the GALNT2 gene and ovarian cancer risk in four population based case-control studies The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Terry, Kathryn L., Allison F. Vitonis, Dena Hernandez, Galina Lurie, Honglin Song et al. 2010. A polymorphism in the GALNT2 gene and ovarian cancer risk in four population based case-control studies. The International Journal of Molecular Epidemiology and Genetics 1(4): 272–277. Published Version http://www.ijmeg.org/files/IJMEG1006002.pdf Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:27337053 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA Int J Mol Epidemiol Genet 2010;1(4):272-277 www.ijmeg.org /IJMEG1006002 Original Article A polymorphism in the GALNT2 gene and ovarian cancer risk in four population based case-control studies Kathryn L Terry1, Allison F Vitonis1, Dena Hernandez2, Galina Lurie3, Honglin Song4, Susan J Ramus5, Linda Titus-Ernstoff6, Michael E Carney3, Lynne R Wilkens3, Aleksandra Gentry-Maharaj5, Usha Menon5, Simon A Gayther5, Paul D Pharaoh4, Marc T Goodman3, Daniel W Cramer1 and Michael J Birrer7 on behalf of the Ovarian Cancer Association Consortium 1Obstetrics and Gynecology Epidemiology Center, Department of Obstetrics and Gynecology, Brigham and Women’s Hospital,
    [Show full text]
  • GALNT2 Expression Is Reduced in Patients with Type 2 Diabetes: Possible Role of Hyperglycemia
    GALNT2 Expression Is Reduced in Patients with Type 2 Diabetes: Possible Role of Hyperglycemia Antonella Marucci1 , Lazzaro di Mauro2 , Claudia Menzaghi1 , Sabrina Prudente3 , Davide Mangiacotti1 , Grazia Fini1, Giuseppe Lotti2, Vincenzo Trischitta1,4*., Rosa Di Paola1*. 1 Research Unit of Diabetes and Endocrine Diseases, IRCCS ‘‘Casa Sollievo della Sofferenza’’, San Giovanni Rotondo, Italy, 2 Blood Bank IRCCS ‘‘Casa Sollievo della Sofferenza’’, San Giovanni Rotondo, Italy, 3 CSS-Mendel, Rome, Italy, 4 Department of Experimental Medicine, Sapienza University, Rome, Italy Abstract Impaired insulin action plays a major role in the pathogenesis of type 2 diabetes, a chronic metabolic disorder which imposes a tremendous burden to morbidity and mortality worldwide. Unraveling the molecular mechanisms underlying insulin resistance would improve setting up preventive and treatment strategies of type 2 diabetes. Down-regulation of GALNT2, an UDPN-acetyl-alpha-D-galactosamine polypeptideN-acetylgalactosaminyltransferase-2 (ppGalNAc-T2), causes impaired insulin signaling and action in cultured human liver cells. In addition, GALNT2 mRNA levels are down-regulated in liver of spontaneously insulin resistant, diabetic Goto-Kakizaki rats. To investigate the role of GALNT2 in human hyperglycemia, we measured GALNT2 mRNA expression levels in peripheral whole blood cells of 84 non-obese and 46 obese non-diabetic individuals as well as of 98 obese patients with type 2 diabetes. We also measured GALNT2 mRNA expression in human U937 cells cultured under different glucose concentrations. In vivo studies indicated that GALNT2 mRNA levels were significantly reduced from non obese control to obese non diabetic and to obese diabetic individuals (p,0.001). In vitro studies showed that GALNT2 mRNA levels was reduced in U937 cells exposed to high glucose concentrations (i.e.
    [Show full text]
  • Analysis of Candidate Idarubicin Drug Resistance Genes in MOLT-3 Cells Using Exome Nuclear DNA
    G C A T T A C G G C A T genes Article Analysis of Candidate Idarubicin Drug Resistance Genes in MOLT-3 Cells Using Exome Nuclear DNA Tomoyoshi Komiyama 1,* ID , Atsushi Ogura 2 ID , Takehito Kajiwara 2 ID , Yoshinori Okada 3 and Hiroyuki Kobayashi 1,* 1 Department of Clinical Pharmacology, Tokai University School of Medicine, Kanagawa 259-1193, Japan 2 Nagahama Institute of Bio-Science and Technology, Shiga 526-0829, Japan; [email protected] (A.O.); [email protected] (T.Ka.) 3 Support Center for Medical Research and Education, Tokai University, Kanagawa 259-1193, Japan; [email protected] * Correspondence: [email protected] (T.Ko.); [email protected] (H.K.); Tel.: +81-463-93-1121 (T.Ko. & H.K.) Received: 18 May 2018; Accepted: 16 July 2018; Published: 1 August 2018 Abstract: Various gene alterations related to acute leukemia are reported to be involved in drug resistance. We investigated idarubicin (IDR) resistance using exome nuclear DNA analyses of the human acute leukemia cell line MOLT-3 and the derived IDR-resistant cell line MOLT-3/IDR. We detected mutations in MOLT-3/IDR and MOLT-3 using both Genome Analysis Toolkit (GATK) and SnpEff program. We found 8839 genes with specific mutations in MOLT-3/IDR and 1162 genes with accompanying amino acid mutations. The 1162 genes were identified by exome analysis of polymerase-related genes using Kyoto Encyclopedia of Genes and Genomes (KEGG) and, among these, we identified genes with amino acid changes.
    [Show full text]
  • GALNT2 (NM 004481) Human Untagged Clone Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for SC314049 GALNT2 (NM_004481) Human Untagged Clone Product data: Product Type: Expression Plasmids Product Name: GALNT2 (NM_004481) Human Untagged Clone Tag: Tag Free Symbol: GALNT2 Synonyms: CDG2T; GalNAc-T2 Vector: pCMV6-XL5 E. coli Selection: Ampicillin (100 ug/mL) Cell Selection: None Fully Sequenced ORF: >OriGene sequence for NM_004481 edited GGGATATCGTCGACCCACGCGTCCGCCACCGCGCCCGCGGCCGGCCCAGGCAGCACTCGC GAGCAGCGGCGGCCCCGCCGGCGGCCGAGTTGGGAGAATGCGGCGGCGCTCGCGGATGCT GCTCTGCTTCGCCTTCCTGTGGGTGCTGGGCATCGCCTACTACATGTACTCGGGGGGCGG CTCTGCGCTGGCCGGGGGCGCGGGCGGCGGCGCCGGCAGGAAGGAGGACTGGAATGAAAT TGACCCCATTAAAAAGAAAGACCTTCATCACAGCAATGGAGAAGAGAAAGCACAAAGCAT GGAGACCCTCCCTCCAGGGAAAGTACGGTGGCCAGACTTTAACCAGGAAGCTTATGTTGG AGGGACGATGGTCCGCTCCGGGCAGGACCCTTACGCCCGCAACAAGTTCAACCAGGTGGA GAGTGATAAGCTTCGAATGGACAGAGCCATCCCTGACACCCGGCATGACCAGTGTCAGCG GAAGCAGTGGCGGGTGGATCTGCCGGCCACCAGCGTGGTGATCACGTTTCACAATGAAGC CAGGTCGGCCCTACTCAGGACCGTGGTCAGCGTGCTTAAGAAAAGCCCGCCCCATCTCAT AAAAGAAATCATCTTGGTGGATGACTACAGCAATGATCCTGAGGACGGGGCTCTCTTGGG GAAAATTGAGAAAGTGCGAGTTCTTAGAAATGATCGACGAGAAGGCCTCATGCGCTCACG GGTTCGGGGGGCCGATGCTGCCCAAGCCAAGGTCCTGACCTTCCTGGACAGTCACTGCGA GTGTAATGAGCACTGGCTGGAGCCCCTCCTGGAAAGGGTGGCGGAGGACAGGACTCGGGT TGTGTCACCCATCATCGATGTCATTAATATGGACAACTTTCAGTATGTGGGGGCATCTGC TGACTTGAAGGGCGGTTTTGATTGGAACTTGGTATTCAAGTGGGATTACATGACGCCTGA GCAGAGAAGGTCCCGGCAGGGGAACCCAGTCGCCCCTATAAAAACCCCCATGATTGCTGG
    [Show full text]
  • Application of Biclustering of Gene Expression Data and Gene Set Enrichment Analysis Methods to Identify Potentially Disease Causing Nanomaterials
    Application of biclustering of gene expression data and gene set enrichment analysis methods to identify potentially disease causing nanomaterials Andrew Williams* and Sabina Halappanavar Full Research Paper Open Access Address: Beilstein J. Nanotechnol. 2015, 6, 2438–2448. Environmental Health Science and Research Bureau, Environmental doi:10.3762/bjnano.6.252 and Radiation Health Sciences Directorate, Health Canada, Ottawa K1A 0K9, Canada Received: 19 August 2015 Accepted: 30 November 2015 Email: Published: 21 December 2015 Andrew Williams* - [email protected] This article is part of the Thematic Series "Nanoinformatics for * Corresponding author environmental health and biomedicine". Keywords: Guest Editor: Y. Cohen gene expression; risk assessment; toxicogenomics © 2015 Williams and Halappanavar; licensee Beilstein-Institut. License and terms: see end of document. Abstract Background: The presence of diverse types of nanomaterials (NMs) in commerce is growing at an exponential pace. As a result, human exposure to these materials in the environment is inevitable, necessitating the need for rapid and reliable toxicity testing methods to accurately assess the potential hazards associated with NMs. In this study, we applied biclustering and gene set enrich- ment analysis methods to derive essential features of altered lung transcriptome following exposure to NMs that are associated with lung-specific diseases. Several datasets from public microarray repositories describing pulmonary diseases in mouse models following exposure to a variety of substances were examined and functionally related biclusters of genes showing similar expres- sion profiles were identified. The identified biclusters were then used to conduct a gene set enrichment analysis on pulmonary gene expression profiles derived from mice exposed to nano-titanium dioxide (nano-TiO2), carbon black (CB) or carbon nanotubes (CNTs) to determine the disease significance of these data-driven gene sets.
    [Show full text]