DOCSLIB.ORG

(RAI14) Promotes Mtor-Mediated Inflammation Under Inflammatory Stress and Chemical Hypoxia in a U87 Glioblastoma Cell Line

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(RAI14) Promotes Mtor-Mediated Inflammation Under Inflammatory Stress and Chemical Hypoxia in a U87 Glioblastoma Cell Line Cellular and Molecular Neurobiology (2019) 39:241–254 https://doi.org/10.1007/s10571-018-0644-z ORIGINAL RESEARCH Retinoic Acid-Induced Protein 14 (RAI14) Promotes mTOR-Mediated Inflammation Under Inflammatory Stress and Chemical Hypoxia in a U87 Glioblastoma Cell Line XiaoGang Shen1,2 · JiaRui Zhang1,2 · XiaoLong Zhang1,2 · YiFan Wang1,2 · YunFeng Hu1,2 · Jun Guo1,2,3 Received: 25 August 2018 / Accepted: 8 December 2018 / Published online: 15 December 2018 © Springer Science+Business Media, LLC, part of Springer Nature 2018 Abstract Retinoic acid-induced 14 is a developmentally regulated gene induced by retinoic acid and is closely associated with NIK/ NF-κB signaling. In the present study, we examined the effect of RAI14 on mTOR-mediated glial inflammation in response to inflammatory factors and chemical ischemia. A U87 cell model of LPS- and TNF-α-induced inflammation was used to investigate the role of RAI14 in glial inflammation. U87 cells were treated with siR-RAI14 or everolimus to detect the correlation between mTOR, RAI14, and NF-κB. CoCl2-stimulated U87 cells were used to analyze the effect of RAI14 on mTOR-mediated NF-κB inflammatory signaling under chemical hypoxia. LPS and TNF-α stimulation resulted in the upregulation of RAI14 mRNA and protein levels in a dose- and time-dependent manner. RAI14 knockdown significantly attenuated the level of pro-inflammatory cytokine via inhibiting the IKK/NF-κB pathway. Treatment with an mTOR inhibitor (everolimus) ameliorated NF-κB activity and IKKα/β phosphorylation via RAI14 signaling. Notably, RAI14 also enhanced mTOR-mediated NF-κB activation under conditions of chemical hypoxia. These findings provide significant insight into the role of RAI14 in mTOR-induced glial inflammation, which is closely associated with infection and ischemia stimuli. Thus, RAI14 may be a potential drug target for the treatment of inflammatory diseases. Keywords RAI14 · Neuroinflmamation · mTOR · NF-κB · Chemical hypoxia Abbreviations 4EBP1 Eukaryotic initiation factor 4E binding protein1 CNS Central nervous system CoCl2 Cobalt chloride XiaoGang Shen and JiaRui Zhang have contributed equally to this work. Co-IP Co-immunoprecipitation eIF4E Eukaryotic translational initiation factor 4 Electronic supplementary material The online version of this epsilon article (https ://doi.org/10.1007/s1057 1-018-0644-z) contains Evero Everolimus supplementary material, which is available to authorized users. * Jun Guo 1 State Key Laboratory Cultivation Base For TCM Quality [email protected] and Efficacy, School of Medicine and Life Science, Nanjing University of Chinese Medicine, Nanjing 210023, XiaoGang Shen People’s Republic of China [email protected] 2 Key Laboratory of Drug Target and Drug for Degenerative JiaRui Zhang Disease, Nanjing University of Chinese Medicine, [email protected] Nanjing 210023, People’s Republic of China XiaoLong Zhang 3 Department of Biochemistry and Molecular Biology, [email protected] Jiangsu Key Laboratory of Therapeutic Material of Chinese YiFan Wang Medicine, School of Medicine and Life Science, Nanjing [email protected] University of Chinese Medicine, Nanjing 210023, People’s Republic of China YunFeng Hu [email protected] Vol.:(0123456789)1 3 242 Cellular and Molecular Neurobiology (2019) 39:241–254 IκB-α NF-kappa-B inhibitor alpha the activation of the mTOR/IκB-α/NF-κB pathway (Temiz- IKK Nuclear factor NF-kappa-B inhibitor kinase Resitoglu et al. 2017). Moreover, the mammalian target of LPS Lipopolysaccharide rapamycin (mTOR) functions as a serine/threonine pro- mRNA Messenger RNA tein kinase and controls protein synthesis, contributing mTOR Mammalian target of rapamycin to the associated physiological and pathological outcome mTORC Mammalian target of rapamycin complex (Caccamo et al. 2018; Wendel et al. 2004). mTOR is com- NF-κB Nuclear factor kappa-light-chain-enhancer of monly comprised of mTOR complexes 1 (mTORC1) and activated B cells 2 (mTORC2). In addition, mTORC1 activity is primarily NIK NF-kappa-B-inducing kinase inhibited by everolimus (RAD001) (Han et al. 2016; Yun PAMPs Pathogen-associated molecular patterns et al. 2016), which has been shown to effectively treat can- PBS Phosphate-buffered saline cers and autoimmune diseases (Kingwell 2013; Vitiello et al. RAI14 Retinoic acid-induced protein 14 2015). Nevertheless, the precise molecular mechanisms Raptor Regulatory-associated protein of mTOR underlying mTORC1-mediated inflammatory stress have RT-qPCR Quantitative real-time polymerase chain yet to be sufficiently identified. reaction In the present study, we have explored the mechanisms si-NC Control siRNA. underlying the RAI14 in mTOR-mediated inflammation siR-RAI14 RAI14 siRNA using models of inflammation and chemical hypoxia. siRNA Small interfering RNA TLRs Toll-like receptors TNF-α Tumor-necrosis factor-α Methods Cell Culture and Treatment Introduction The human glioma cell (U87) or human umbilical vein The human novel retinal pigment epithelial cell gene (NOR- endothelial cell (HUVEC) was obtained from the American PEG, RAI14) is a developmentally regulated gene originally Type Culture Collection (ATCC, Manassas, VA, USA). U87 derived from the human retinal pigment epithelial cell line, and HUVEC cells were cultured in Dulbecco’s Modification ARPE-19 (Kutty et al. 2001). This gene encodes the RAI14 of Eagle’s Medium (DMEM; Gibco, Carlsbad, CA, USA) protein that contains six ankyrin repeats and an extended supplemented with 10% (v/v) fetal bovine serum (Gibco), coiled-coil domain. Moreover, RAI14 is widely expressed 100 U/mL penicillin and 100 µg/mL streptomycin (Gibco). in a variety of human tissues and participates in the reor- The cells were cultured in 60-mm culture dishes at 37 °C in a ganization of actin filaments in the testis, contributing to humidified atmosphere of 5%CO 2. U87 cells were grown on spermatid polarity and cell adhesion (Chen et al. 2018a, b; 60-mm culture dishes then incubated with 20 nM everolimus Peng et al. 2000; Qian et al. 2013). While the Gene Ontol- (MedChem Express, Monmouth Junction, NJ, USA) with ogy has deduced that RAI14 is associated with NIK/NF-κB or without TNF-α, LPS, or CoCl 2 as a baseline for further signaling (Huttlin et al. 2017), the function of RAI14 in glial research. Recombinant human TNF-α was obtained from inflammation remains unknown. Hangzhou Biologic Biotechnology (China). All cell experi- It has been well established that the NF-κB signaling ments were repeated at least three times. pathway is involved in the wide variety of pathological conditions, such as cancer and neurodegenerative diseases Antibodies (Collignon et al. 2018; Li et al. 2012). Under different patho- logical conditions (e.g., LPS, TNF-α and hypoxia), NF-κB Rabbit anti-RAI14 polyclonal antibody, rabbit anti-GAPDH p65 activation and nuclear translocation control various tran- polyclonal antibody, mouse anti-α-tubulin monoclonal anti- scriptional responses (Li et al. 2016). Thus, pro-inflamma- body, rabbit anti-phospho-mTOR (Ser2448) antibody and tory agonists, such as LPS and TNF-α, could play a vital role rabbit anti-phospho-eIF4EBP1 (Thr37 + Thr46) antibody in the progression of diseases of the central nervous system. were purchased from Bioss (Beijing, China). Phospho- Additionally, since hypoxia can trigger a pro-inflammatory IKKα/β (Ser176/180) antibody and NF-κB p65 rabbit mon- immune response (Cosin-Roger et al. 2017; Subhan et al. oclonal antibody were obtained from Beyotime (Shanghai, 2017), CoCl2 (cobalt chloride), a hypoxia-mimetic agent, China). Rabbit anti-α-actin monoclonal antibody, rabbit can be used to make an analogical analysis of the NF-κB anti-β-actin polyclonal antibody, and rabbit anti-β-tubulin inflammatory signaling pathway under conditions of chemi- monoclonal antibody were obtained from BOSTER (Wuhan, cal hypoxia. It has previously been shown that mTOR is China). All secondary antibodies were purchased from associated with proinflammatory immune responses through ZSGB-BIO (Beijing, China). 1 3 Cellular and Molecular Neurobiology (2019) 39:241–254 243 siRNA Transfection was evaluated using an ECL Western Bot analysis kit (Mil- lipore) and were normalized to those of α-actin or β-actin or The siRNA probe s25049 for RAI14, siRNA probe s25048 α-tubulin or β-tubulin or GAPDH. for RAI14 (Hasson et al. 2013), GAPDH positive control oligonucleotides and its control small interfering RNA Co‑immunoprecipitation (siRNA) in experiments refer to an All-Starnon silencing siRNA synthesized by Shanghai GenePharma (Shanghai, According to the manufacturer’s instructions, 2 µg NF-κB China). Lipo6000™ Transfection reagent (Beyotime) was p65 rabbit monoclonal antibody was added to the same used for the transfection of siRNA according to the manu- amount (500 µg) of cell lysate overnight at 4 °C. Protein facturer’s instructions. A/G-agarose spheres (Santa Cruz Biotechnology, Dallas, TX, USA) were added and the samples stored at 4 °C for 2 h. The samples were then centrifuged at 13,000g at 4 °C for Quantitative Real‑Time Polymerase Chain Reaction 1 min, washed three times with lysis buffer, and boiled for 10 min after the addition of 30 µL 1× SDS Loading Buffer. A Reverse Transcription Kit and the SYBR Green PCR Mas- The denatured samples were kept at − 20 °C for future west- ter Mix were obtained from Takara (Otsu, Shiga, Japan). ern blotting. Total RNA was isolated from the cells using Trizol reagent according to the manufacturer’s instructions (Beyotime) Immunofluorescence Analysis and cDNA was reverse-transcribed using a Reverse
Load more

Recommended publications
  • Downregulation of RAI14 Inhibits the Proliferation
    Journal of Cancer 2019, Vol. 10 6341 Ivyspring International Publisher Journal of Cancer 2019; 10(25): 6341- 6348. doi: 10.7150/jca.34910 Research Paper Downregulation of RAI14 inhibits the proliferation and invasion of breast cancer cells Ming Gu1, Wenhui Zheng2, Mingdi Zhang3, Xiaoshen Dong1, Yan Zhao1, Shuo Wang1, Haiyang Jiang1, Lu Liu1, Xinyu Zheng1,4 1. Department of Breast Surgery, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, People’s Republic of China 2. Department of anesthesiology, The Shengjing Hospital of China Medical University, Shenyang, Liaoning 110001, People’s Republic of China 3. Department of Breast Surgery, Obstetrics and Gynecology Hospital of Fudan University, Shanghai, 200011, People’s Republic of China 4. Lab 1, Cancer Institute, The First Affiliated Hospital of China Medical University, Shenyang, Liaoning 110001, People’s Republic of China Corresponding author: Prof. Xinyu Zheng, Department of Breast Surgery, The First Affiliated Hospital of China Medical University, 155 Nanjing North Street, Heping District, Shenyang City, Liaoning Province, 110001, People’s Republic of China. Fax number: 0086 24 83282741 Telephone number: 0086 24 83282741 E-mail: [email protected] © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2019.08.26; Accepted: 2019.09.19; Published: 2019.10.18 Abstract Retinoic acid-induced 14 (RAI14) is involved in the development of different tumor types, however, its expression and biological function in breast cancer are yet unknown.
    [Show full text]
  • Exon-Based Clustering of Murine Breast Tumor Transcriptomes Reveals Alternative Exons Whose Expression Is Associated with Metastasis
    Published OnlineFirst January 26, 2010; DOI: 10.1158/0008-5472.CAN-09-2703 Published Online First on January 26, 2010 as 10.1158/0008-5472.CAN-09-2703 Integrated Systems and Technologies Cancer Research Exon-Based Clustering of Murine Breast Tumor Transcriptomes Reveals Alternative Exons Whose Expression Is Associated with Metastasis Martin Dutertre1, Magali Lacroix-Triki3,4,5, Keltouma Driouch6, Pierre de la Grange2, Lise Gratadou1, Samantha Beck3,4,5, Stefania Millevoi3,4,5, Jamal Tazi7, Rosette Lidereau6, Stephan Vagner3,4,5, and Didier Auboeuf1 Abstract In the field of bioinformatics, exon profiling is a developing area of disease-associated transcriptome anal- ysis. In this study, we performed a microarray-based transcriptome analysis at the single exon level in mouse 4T1 primary mammary tumors with different metastatic capabilities. A novel bioinformatics platform was developed that identified 679 genes with differentially expressed exons in 4T1 tumors, many of which were involved in cell morphology and movement. Of 152 alternative exons tested by reverse transcription-PCR, 97 were validated as differentially expressed in primary tumors with different metastatic capability. This anal- ysis revealed candidate progression genes, hinting at variations in protein functions by alternate exon usage. In a parallel effort, we developed a novel exon-based clustering analysis and identified alternative exons in tumor transcriptomes that were associated with dissemination of primary tumor cells to sites of pulmonary metastasis. This analysis also revealed that the splicing events identified by comparing primary tumors were not aberrant events. Lastly, we found that a subset of differentially spliced variant transcripts identified in the murine model was associated with poor prognosis in a large clinical cohort of patients with breast cancer.
    [Show full text]
  • Stem Cells® Original Article
    ® Stem Cells Original Article Properties of Pluripotent Human Embryonic Stem Cells BG01 and BG02 XIANMIN ZENG,a TAKUMI MIURA,b YONGQUAN LUO,b BHASKAR BHATTACHARYA,c BRIAN CONDIE,d JIA CHEN,a IRENE GINIS,b IAN LYONS,d JOSEF MEJIDO,c RAJ K. PURI,c MAHENDRA S. RAO,b WILLIAM J. FREEDa aCellular Neurobiology Research Branch, National Institute on Drug Abuse, Department of Health and Human Services (DHHS), Baltimore, Maryland, USA; bLaboratory of Neuroscience, National Institute of Aging, DHHS, Baltimore, Maryland, USA; cLaboratory of Molecular Tumor Biology, Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA; dBresaGen Inc., Athens, Georgia, USA Key Words. Embryonic stem cells · Differentiation · Microarray ABSTRACT Human ES (hES) cell lines have only recently been compared with pooled human RNA. Ninety-two of these generated, and differences between human and mouse genes were also highly expressed in four other hES lines ES cells have been identified. In this manuscript we (TE05, GE01, GE09, and pooled samples derived from describe the properties of two human ES cell lines, GE01, GE09, and GE07). Included in the list are genes BG01 and BG02. By immunocytochemistry and reverse involved in cell signaling and development, metabolism, transcription polymerase chain reaction, undifferenti- transcription regulation, and many hypothetical pro- ated cells expressed markers that are characteristic of teins. Two focused arrays designed to examine tran- ES cells, including SSEA-3, SSEA-4, TRA-1-60, TRA-1- scripts associated with stem cells and with the 81, and OCT-3/4. Both cell lines were readily main- transforming growth factor-β superfamily were tained in an undifferentiated state and could employed to examine differentially expressed genes.
    [Show full text]
  • Cytogenetic and Molecular Characterization of the Macro- And
    University of Ulm Department of Human Genetics Prof. Dr. med. Walther Vogel Cytogenetic and Molecular Characterization of the Macro- and Micro-inversions, which Distinguish the Human and the Chimpanzee Karyotypes - from Speciation to Polymorphism Thesis Applying for the Degree of Doctor of Human Biology (Dr. hum. biol.) Faculty of Medicine University of Ulm Presented by Justyna Monika Szamalek from Wrze śnia in Poland 2006 Amtierender Dekan: Prof. Dr. Klaus-Michael Debatin 1. Berichterstatter: Prof. Dr. med. Horst Hameister 2. Berichterstatter: Prof. Dr. med. Konstanze Döhner Tag der Promotion: 28.07.2006 Content Content 1. Introduction ...................................................................................................................7 1.1. Primate phylogeny........................................................................................................7 1.2. Africa as the place of human origin and the living area of the present-day chimpanzee populations .................................................................9 1.3. Cytogenetic and molecular differences between human and chimpanzee genomes.............................................................................................10 1.4. Cytogenetic and molecular differences between common chimpanzee and bonobo genomes................................................................................17 1.5. Theory of speciation .....................................................................................................18 1.6. Theory of selection
    [Show full text]
  • HSP90-Incorporating Chaperome Networks As Biosensor for Disease-Related Pathways in Patient- Specific Midbrain Dopamine Neurons
    ARTICLE DOI: 10.1038/s41467-018-06486-6 OPEN HSP90-incorporating chaperome networks as biosensor for disease-related pathways in patient- specific midbrain dopamine neurons Sarah Kishinevsky1,2,3,4, Tai Wang3, Anna Rodina3, Sun Young Chung1,2, Chao Xu3, John Philip5, Tony Taldone3, Suhasini Joshi3, Mary L. Alpaugh3,14, Alexander Bolaender3, Simon Gutbier6, Davinder Sandhu7, Faranak Fattahi1,2, Bastian Zimmer1,2, Smit K. Shah3, Elizabeth Chang5, Carmen Inda3,15, John Koren 3rd3,16, Nathalie G. Saurat1,2, Marcel Leist 6, Steven S. Gross7, Venkatraman E. Seshan8, Christine Klein9, Mark J. Tomishima1,2,10, Hediye Erdjument-Bromage11,12, Thomas A. Neubert 11,12, Ronald C. Henrickson5, 1234567890():,; Gabriela Chiosis3,13 & Lorenz Studer1,2 Environmental and genetic risk factors contribute to Parkinson’s Disease (PD) pathogenesis and the associated midbrain dopamine (mDA) neuron loss. Here, we identify early PD pathogenic events by developing methodology that utilizes recent innovations in human pluripotent stem cells (hPSC) and chemical sensors of HSP90-incorporating chaperome networks. We show that events triggered by PD-related genetic or toxic stimuli alter the neuronal proteome, thereby altering the stress-specific chaperome networks, which produce changes detected by chemical sensors. Through this method we identify STAT3 and NF-κB signaling activation as examples of genetic stress, and phospho-tyrosine hydroxylase (TH) activation as an example of toxic stress-induced pathways in PD neurons. Importantly, pharmacological inhibition of the stress chaperome network reversed abnormal phospho- STAT3 signaling and phospho-TH-related dopamine levels and rescued PD neuron viability. The use of chemical sensors of chaperome networks on hPSC-derived lineages may present a general strategy to identify molecular events associated with neurodegenerative diseases.
    [Show full text]
  • Transdifferentiation of Human Mesenchymal Stem Cells
    Transdifferentiation of Human Mesenchymal Stem Cells Dissertation zur Erlangung des naturwissenschaftlichen Doktorgrades der Julius-Maximilians-Universität Würzburg vorgelegt von Tatjana Schilling aus San Miguel de Tucuman, Argentinien Würzburg, 2007 Eingereicht am: Mitglieder der Promotionskommission: Vorsitzender: Prof. Dr. Martin J. Müller Gutachter: PD Dr. Norbert Schütze Gutachter: Prof. Dr. Georg Krohne Tag des Promotionskolloquiums: Doktorurkunde ausgehändigt am: Hiermit erkläre ich ehrenwörtlich, dass ich die vorliegende Dissertation selbstständig angefertigt und keine anderen als die von mir angegebenen Hilfsmittel und Quellen verwendet habe. Des Weiteren erkläre ich, dass diese Arbeit weder in gleicher noch in ähnlicher Form in einem Prüfungsverfahren vorgelegen hat und ich noch keinen Promotionsversuch unternommen habe. Gerbrunn, 4. Mai 2007 Tatjana Schilling Table of contents i Table of contents 1 Summary ........................................................................................................................ 1 1.1 Summary.................................................................................................................... 1 1.2 Zusammenfassung..................................................................................................... 2 2 Introduction.................................................................................................................... 4 2.1 Osteoporosis and the fatty degeneration of the bone marrow..................................... 4 2.2 Adipose and bone
    [Show full text]
  • Genome-Wide Association Studies for Sperm Traits in Assaf Sheep Breed
    ANIMAL-100065; No of Pages 9 Animal xxx (2021) xxx Contents lists available at ScienceDirect Animal The international journal of animal biosciences Genome-wide association studies for sperm traits in Assaf sheep breed M. Serrano a,⁎, M. Ramón b, J.H. Calvo c,f, M.Á. Jiménez a, F. Freire d, J.M. Vázquez d, J.J. Arranz e a Departamento de Mejora Genética Animal, INIA, 28040 Madrid, Spain b IRIAF-CERSYRA, Valdepeñas 13300, Ciudad Real, Spain c Unidad de Tecnología en Producción Animal, CITA, 59059 Zaragoza, Spain d OVIGEN, Granja Florencia s/n, Ctra. Villalazán-Peleagonzalo, 49800 Toro, Zamora, Spain e Departamento de Producción Animal, Universidad de León, 24007 León, Spain f ARAID, 50004 Zaragoza, Spain article info abstract Article history: Sperm quality traits routinely collected by artificial insemination (AI) center for rams progeny test are related Received 6 May 2020 with the capacity to produce sperm doses for AI and, in more or less grade, with males' fertility. Low-quality ejac- Received in revised form 20 August 2020 ulates are unuseful to perform AI sperm doses, which suppose high economic loses for the AI center. Moreover, Accepted 21 August 2020 sperm quality traits have low heritability values which make traditional genetic selection little efficient to its im- Available online xxxx provement. In this work, a genome-wide association study (GWAS) was conducted by using sperm quality traits data and 50 K Affymetrix custom chip genotypes of 429 rams of Assaf breed from OVIGEN AI centre. Furthermore, Keywords: Association study 47 of these rams were also genotyped with the Illumina HD Ovine BeadChip, and therefore HD genotypes were Pseudo-phenotypes imputed for all rams with phenotype data.
    [Show full text]
  • Table S1. 103 Ferroptosis-Related Genes Retrieved from the Genecards
    Table S1. 103 ferroptosis-related genes retrieved from the GeneCards. Gene Symbol Description Category GPX4 Glutathione Peroxidase 4 Protein Coding AIFM2 Apoptosis Inducing Factor Mitochondria Associated 2 Protein Coding TP53 Tumor Protein P53 Protein Coding ACSL4 Acyl-CoA Synthetase Long Chain Family Member 4 Protein Coding SLC7A11 Solute Carrier Family 7 Member 11 Protein Coding VDAC2 Voltage Dependent Anion Channel 2 Protein Coding VDAC3 Voltage Dependent Anion Channel 3 Protein Coding ATG5 Autophagy Related 5 Protein Coding ATG7 Autophagy Related 7 Protein Coding NCOA4 Nuclear Receptor Coactivator 4 Protein Coding HMOX1 Heme Oxygenase 1 Protein Coding SLC3A2 Solute Carrier Family 3 Member 2 Protein Coding ALOX15 Arachidonate 15-Lipoxygenase Protein Coding BECN1 Beclin 1 Protein Coding PRKAA1 Protein Kinase AMP-Activated Catalytic Subunit Alpha 1 Protein Coding SAT1 Spermidine/Spermine N1-Acetyltransferase 1 Protein Coding NF2 Neurofibromin 2 Protein Coding YAP1 Yes1 Associated Transcriptional Regulator Protein Coding FTH1 Ferritin Heavy Chain 1 Protein Coding TF Transferrin Protein Coding TFRC Transferrin Receptor Protein Coding FTL Ferritin Light Chain Protein Coding CYBB Cytochrome B-245 Beta Chain Protein Coding GSS Glutathione Synthetase Protein Coding CP Ceruloplasmin Protein Coding PRNP Prion Protein Protein Coding SLC11A2 Solute Carrier Family 11 Member 2 Protein Coding SLC40A1 Solute Carrier Family 40 Member 1 Protein Coding STEAP3 STEAP3 Metalloreductase Protein Coding ACSL1 Acyl-CoA Synthetase Long Chain Family Member 1 Protein
    [Show full text]
  • High Expression of the Sd Synthase B4GALNT2 Associates with Good
    cells Article High Expression of the Sda Synthase B4GALNT2 Associates with Good Prognosis and Attenuates Stemness in Colon Cancer Michela Pucci y, Inês Gomes Ferreira y, Martina Orlandani, Nadia Malagolini, Manuela Ferracin and Fabio Dall’Olio * Department of Experimental, Diagnostic and Specialty Medicine (DIMES), General Pathology Building, University of Bologna, Via San Giacomo 14, Via San Giacomo 14, 40126 Bologna, Italy; [email protected] (M.P.); [email protected] (I.G.F.); [email protected] (M.O.); [email protected] (N.M.); [email protected] (M.F.) * Correspondence: [email protected]; Tel.: +39-051-2094704 These authors have equal contribution. y Received: 25 February 2020; Accepted: 7 April 2020; Published: 11 April 2020 Abstract: Background: The carbohydrate antigen Sda and its biosynthetic enzyme B4GALNT2 are highly expressed in normal colonic mucosa but are down-regulated to a variable degree in colon cancer tissues. Here, we investigated the clinical and biological importance of B4GALNT2 in colon cancer. Methods: Correlations of B4GALNT2 mRNA with clinical data were obtained from The Cancer Genome Atlas (TCGA) database; the phenotypic and transcriptomic changes induced by B4GALNT2 were studied in LS174T cells transfected with B4GALNT2 cDNA. Results: TCGA data indicate that patients with high B4GALNT2 expression in cancer tissues display longer survival than non-expressers. In LS174T cells, expression of B4GALNT2 did not affect the ability to heal a scratch wound or to form colonies in standard growth conditions but markedly reduced the growth in soft agar, the tridimensional (3D) growth as spheroids, and the number of cancer stem cells, indicating a specific effect of B4GALNT2 on the growth in poor adherence and stemness.
    [Show full text]
  • Autocrine IFN Signaling Inducing Profibrotic Fibroblast Responses by a Synthetic TLR3 Ligand Mitigates
    Downloaded from http://www.jimmunol.org/ by guest on September 28, 2021 Inducing is online at: average * The Journal of Immunology published online 16 August 2013 from submission to initial decision 4 weeks from acceptance to publication http://www.jimmunol.org/content/early/2013/08/16/jimmun ol.1300376 A Synthetic TLR3 Ligand Mitigates Profibrotic Fibroblast Responses by Autocrine IFN Signaling Feng Fang, Kohtaro Ooka, Xiaoyong Sun, Ruchi Shah, Swati Bhattacharyya, Jun Wei and John Varga J Immunol Submit online. Every submission reviewed by practicing scientists ? is published twice each month by http://jimmunol.org/subscription Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://www.jimmunol.org/content/suppl/2013/08/20/jimmunol.130037 6.DC1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2013 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of September 28, 2021. Published August 16, 2013, doi:10.4049/jimmunol.1300376 The Journal of Immunology A Synthetic TLR3 Ligand Mitigates Profibrotic Fibroblast Responses by Inducing Autocrine IFN Signaling Feng Fang,* Kohtaro Ooka,* Xiaoyong Sun,† Ruchi Shah,* Swati Bhattacharyya,* Jun Wei,* and John Varga* Activation of TLR3 by exogenous microbial ligands or endogenous injury-associated ligands leads to production of type I IFN.
    [Show full text]
  • Intragenic and Large NIPBL Rearrangements Revealed by MLPA in Cornelia De Lange Patients
    European Journal of Human Genetics (2012) 20, 734–741 & 2012 Macmillan Publishers Limited All rights reserved 1018-4813/12 www.nature.com/ejhg ARTICLE Intragenic and large NIPBL rearrangements revealed by MLPA in Cornelia de Lange patients Silvia Russo*,1, Maura Masciadri1, Cristina Gervasini2, Jacopo Azzollini2, Anna Cereda3, Giuseppe Zampino4, Oskar Haas5, Gioacchino Scarano6, Maja Di Rocco7, Palma Finelli1, Romano Tenconi8, Angelo Selicorni3 and Lidia Larizza1,2 Cornelia de Lange syndrome (CdLS) is a rare multisystemic congenital anomaly disorder that is characterised by intellectual disability and growth retardation, congenital heart defects, intestinal anomalies, facial dysmorphism (including synophyris and high arched eyebrows) and limb reduction defects. Mutations in three cohesin-associated genes encoding a key regulator (NIPBL, chr 5p13.2) and one structural component of the cohesin ring (SMC1A, chr Xp11) occur in about 65% of CdLS patients. NIPBL is the major causative gene, and accounts for 40–60% of CdLS patients as shown by a number of mutational screening studies that indicate a wide mutational repertoire of mainly small deletions and point mutations. Only a few data are available concerning the occurrence of large NIPBL rearrangements or intragenic deletions or duplications involving whole exons. We used multiplex ligation-dependent probe amplification (MLPA) to study 132 CdLS patients negative to the standard mutation NIPBL test out of a cohort of 200 CdLS patients. A total of 7 out of 132 patients were found to carry NIPBL alterations, including two large gene deletions extending beyond the gene, four intragenic multi- or single-exon deletions and one single- exon duplication. These findings show that MLPA leads to a 5.3% increase in the detection of mutations when used in addition to the standard NIPBL scan, and contributes per se to the molecular diagnosis of 3.5% (7/200) of clinically diagnosed CdLS patients.
    [Show full text]
  • Rai14 (Retinoic Acid Induced Protein 14) Is Involved in Regulating F-Actin Dynamics at the Ectoplasmic Specialization in the Rat Testis*
    Rai14 (Retinoic Acid Induced Protein 14) Is Involved in Regulating F-Actin Dynamics at the Ectoplasmic Specialization in the Rat Testis* Xiaojing Qian1,2, Dolores D. Mruk1, C. Yan Cheng1* 1 The Mary M. Wohlford Laboratory for Male Contraceptive Research, Center for Biomedical Research, Population Council, New York, New York, United States of America, 2 Department of Anatomy, Histology and Embryology, School of Basic Medicine, Peking Union Medical College, Beijing, China Abstract Rai14 (retinoic acid induced protein 14) is an actin binding protein first identified in the liver, highly expressed in the placenta, the testis, and the eye. In the course of studying actin binding proteins that regulate the organization of actin filament bundles in the ectoplasmic specialization (ES), a testis-specific actin-rich adherens junction (AJ) type, Rai14 was shown to be one of the regulatory proteins at the ES. In the rat testis, Rai14 was found to be expressed by Sertoli and germ cells, structurally associated with actin and an actin cross-linking protein palladin. Its expression was the highest at the ES in the seminiferous epithelium of adult rat testes, most notably at the apical ES at the Sertoli-spermatid interface, and expressed stage-specifically during the epithelial cycle in stage VII-VIII tubules. However, Rai14 was also found at the basal ES near the basement membrane, associated with the blood-testis barrier (BTB) in stage VIII-IX tubules. A knockdown of Rai14 in Sertoli cells cultured in vitro by RNAi was found to perturb the Sertoli cell tight junction-permeability function in vitro, mediated by a disruption of F-actin, which in turn led to protein mis-localization at the Sertoli cell BTB.
    [Show full text]