DOCSLIB.ORG

Circighg-Induced Epithelial-To-Mesenchymal

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Circighg-Induced Epithelial-To-Mesenchymal Author Manuscript Published OnlineFirst on November 17, 2020; DOI: 10.1158/0008-5472.CAN-20-0554 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 1 Title 2 circIGHG-induced epithelial-to-mesenchymal transition promotes oral squamous cell carcinoma 3 progression via miR-142-5p/IGF2BP3 signaling 4 5 Authors and affiliations 6 Jingpeng Liu, Xiao Jiang, Ailing Zou, Zhaoyi Mai, Zhijie Huang, Liying Sun, Jianjiang Zhao 7 Authors’ affiliation: Department of Oral Surgery, Stomatological Hospital, Southern Medical 8 University, Guangzhou, China. 9 10 Running title 11 circIGHG regulates OSCC via miR-142-5p/IGF2BP3 signaling 12 13 Correspondence: Jianjiang Zhao, Department of Oral Surgery, Stomatological Hospital, 14 Southern Medical University, China. 15 366 S Jiangnan Street, Haizhu District, Guangzhou City, Guangdong Province, China. 16 Tel: +86-020-84439500 17 Mobile: +86-13609645845 18 Email: [email protected] 19 20 Keywords 21 circular RNA, OSCC, EMT, metastasis, miRNA 22 23 Competing interest 24 The authors declare no potential conflicts of interest. 25 26 Abbreviations 27 OSCC, oral squamous cell carcinoma; circRNA, circular RNA; miRNA, microRNA; 28 EMT, epithelial-mesenchymal transition; IGF2BP3, Insulin like growth factor 2 mRNA binding 29 protein 3; IGHG, immunoglobulin heavy chain G; CCCP, carbonyl cyanide 3- 30 chlorophenylhydrazone; FISH, fluorescence in situ hybridization; RT-qPCR, real time 31 quantitative polymerase chain reaction; IHC, immunohistochemistry; IF, immunofluorescence; 32 GEO, Gene Expression Omnibus. 33 34 Word count: 4658 35 Number of figures: 6 36 Number of tables: 0 37 38 39 40 41 42 43 44 45 1 Downloaded from cancerres.aacrjournals.org on October 4, 2021. © 2020 American Association for Cancer Research. Author Manuscript Published OnlineFirst on November 17, 2020; DOI: 10.1158/0008-5472.CAN-20-0554 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 1 Abstract 2 Circular RNAs are a new member of endogenously produced noncoding RNAs that have been 3 characterized as key regulators of gene expression in a variety of malignances. However, the role 4 of circRNA in oral squamous cell carcinoma (OSCC) remains largely unknown. In this study, we 5 identified unique circRNA that regulate OSCC progression and metastasis and pave roads for 6 future research in early diagnosis, prevention, and treatment of OSCC. Transcriptomic analyses 7 identified a circRNA derived from immunoglobulin heavy chain G locus (circIGHG) as 8 significantly upregulated in OSCC and positively associated with poor prognosis of OSCC. 9 circIGHG directly bound miR-142-5p and consequently elevated IGF2BP3 activity. Knockdown 10 of circIGHG led to impaired expression of IGF2BP3 and attenuated aggressiveness of OSCC 11 cells. EMT was the main mechanism through which circIGHG/IGF2BP3 promotes metastasis of 12 OSCC. Overall, these results demonstrate that circIGHG plays pivotal role in OSCC 13 development and metastasis and has potential to serve as a biomarker and therapeutic target for 14 early-stage diagnosis and treatment of OSCC. 15 16 Statement of significance 17 Findings broaden our insights regarding regulation of OSCC progression by circular RNA and 18 serve as a reference for future clinical research in OSCC diagnosis and treatment. 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 2 Downloaded from cancerres.aacrjournals.org on October 4, 2021. © 2020 American Association for Cancer Research. Author Manuscript Published OnlineFirst on November 17, 2020; DOI: 10.1158/0008-5472.CAN-20-0554 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 1 Introduction 2 Oral cancer is overall the eighth most common malignancy and the most prevalent disease in 3 head and neck tumors, with 170,000 deaths and over 350,000 newly added cases in 2018 alone 4 worldwide(1,2). Squamous cell carcinoma accounts for over 95% of oral cancer, which is 5 characterized by robust aggressiveness and frequent metastasis and recurrence(3). Although 6 recent years have witnessed remarkable improvements in diagnosis and treatment, 5-year 7 survival rate of OSCC patients remains under 50% and lack of early detection is responsible for 8 the high mortality (4,5). Therefore, understanding the mechanisms of progression and metastasis 9 of OSCC is an important goal in OSCC intervention. 10 11 Circular RNAs (circRNAs) are a recently re-discovered type of endogenous non-coding RNA 12 that are generated from back-splicing of precursor mRNAs and features distinguishable single- 13 stranded, covalently closed loops, lacking 5’-3’ polarity(6,7). Extensively distributed in 14 cytoplasm, this unique structure makes circRNA more resistant to exonucleases and more stable 15 than their linear RNA counterparts(8). For decades, circRNAs were nearly unexplored due to 16 facts that circRNA-specific back-splicing junction reads do not map to a linear reference genome 17 and circRNAs do not have poly(A) tails and are therefore not detectable in most early RNA-seq 18 datasets where poly(A)-enrichment is part of the library preparation protocol. Over the past few 19 years, circRNAs have gained enormous investigations. Despite controversy and skepticism still 20 hovering over circRNA research realm(9), there are increasing amount of studies showing that 21 circRNAs have great potential as miRNA sponges in a complementary base-pairing manner, 22 suppressing miRNA activity and thereby promoting translation and stability of downstream 23 target genes(10). Such a circRNA/miRNA/mRNA signaling axis has been demonstrated to exert 24 multiple functions in a broad variety of physiological and pathological conditions, particularly in 25 cancers(11-13). Accumulating research has identified numerous non-coding RNAs that 26 participate in OSCC progression regulation, including miR-23a-3p which suppresses tumor 27 growth in OSCC via targeting fibroblast growth factor 2 (FGF-2)(14) and lncRNA-p23154 28 whose inhibitory action over miR-378a-3 facilitates GLUT-1 expression, therefore promoting 29 invasion and metastasis of OSCC(15). With a growing body of reports that indicate emerging 30 roles and functions of circular RNAs in OSCC, how circRNAs pathways delicately regulate 31 OSCC progression still remains largely enigmatic and needs to be further explored. 32 33 Human immunoglobulin heavy chain G (IGHG) gene, located in chromosome region 14q32, is 34 known for polymorphism across populations and critically involved in immune response and 35 human survival(16,17). Although not quite often associated to cancers, dysregulation of IGHG 36 was reported in multiple malignancies, including lung cancer, breast cancer and gastric 37 cancer(18-20). In this study, we identified a unique IGHG-derived circular RNA, 38 hsa_circ_0000579 (termed circIGHG), whose upregulated expression was frequently observed in 39 OSCC and associated with poor prognosis. Further investigations demonstrated that circIGHG 40 facilitates epithelial-mesenchymal transition (EMT) by regulating IGF2BP3 via directly 41 sponging miR-142-5p, thereby leading to increased aggressiveness of OSCC. Those findings 42 endow circIGHG promising potentials in serving as a novel diagnostic and therapeutic marker in 43 OSCC. 44 Materials and Methods 45 RNA isolation, RNase R treatment, and PCR 3 Downloaded from cancerres.aacrjournals.org on October 4, 2021. © 2020 American Association for Cancer Research. Author Manuscript Published OnlineFirst on November 17, 2020; DOI: 10.1158/0008-5472.CAN-20-0554 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. 1 Total RNA was isolated using RNAprep Pure kits (DP430, DP431, Tiangen, China) according to 2 the manufacturer’s guidelines from either liquid nitrogen-frozen tissues or cultured cells. 3 Concentration and purity for each RNA extract was checked by 260/280 ratio on a NanoDrop 4 2000 instrument (Thermofisher, US) using RNA-specific protocol. First strand cDNA synthesis 5 was achieved by using FastKing RT Kit (KR116, Tiangen, China). Quantitative PCR was 6 performed with the Talent qPCR premix Kit (FP209, Tiangen, China) following the product’s 7 instructions. Primers used in this study were designed using Primer Bank 8 (https://pga.mgh.harvard.edu/primerbank/) and Primer3Plus (V2.5.0). Primer sequences were 9 listed in Supplementary file 1: Table S1. Three technical replicates were set for each single 10 reaction to ensure reliability and validity. Reactions were programmed for 1 cycle of 11 denaturation for 1.5 minutes at 95°C, followed by 40 cycles of extension including 30 seconds at 12 95°C, 50 seconds at 57°C and 1.5 minutes at 72°C. The ΔΔCT value of the target genes’ 13 expression was measured and assessed against the value of the reference gene GAPDH. 14 15 RNase R treatment was performed by incubating 2μg of total RNA mixture at 37°C for 10 16 minutes, as suggested by the product’s manual (RNR07250, Lucigen, US). Following removal of 17 linear forms, RT-qPCR qualification was performed to confirm the circular structure of 18 circIGHG. Sanger sequencing covering the back-splicing junction sequence was performed using 19 the RT-qPCR product of circIGHG (Geneseed, China). 1% agarose gel was used for DNA 20 electrophoresis. 21 22 RNA FISH 23 RNA FISH was performed using the Fluorescent in situ Hybridization Kit (C10910, RiboBio, 24 China) according to the manufacturer’s instructions. Cy3-labeled probes targeting circIGHG and 25 Dig-labeled miR-142-5p probes (see Supplementary file 1: Table S2) were synthesized by 26 RiboBio Technology Co. Ltd (China). Fluorescence was excited and recorded with a Zeiss 27 confocal laser scanning microscope (LSM 880 with Airyscan, Carl Zeiss, Germany). 28 29 RNA pull-down assay 30 RNA pull-down assay was performed referring to Lal A’s introduction(21). Biotinylated 31 circIGHG and miR-142-5p probes were synthesized and transfected into CAL-27 cells, 32 according to the manufacturer’s protocol (RiboBio, Guangzhou, China).
Load more

Recommended publications
  • Genetic and Pharmacological Approaches to Preventing Neurodegeneration
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2012 Genetic and Pharmacological Approaches to Preventing Neurodegeneration Marco Boccitto University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Neuroscience and Neurobiology Commons Recommended Citation Boccitto, Marco, "Genetic and Pharmacological Approaches to Preventing Neurodegeneration" (2012). Publicly Accessible Penn Dissertations. 494. https://repository.upenn.edu/edissertations/494 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/494 For more information, please contact [email protected]. Genetic and Pharmacological Approaches to Preventing Neurodegeneration Abstract The Insulin/Insulin-like Growth Factor 1 Signaling (IIS) pathway was first identified as a major modifier of aging in C.elegans. It has since become clear that the ability of this pathway to modify aging is phylogenetically conserved. Aging is a major risk factor for a variety of neurodegenerative diseases including the motor neuron disease, Amyotrophic Lateral Sclerosis (ALS). This raises the possibility that the IIS pathway might have therapeutic potential to modify the disease progression of ALS. In a C. elegans model of ALS we found that decreased IIS had a beneficial effect on ALS pathology in this model. This beneficial effect was dependent on activation of the transcription factor daf-16. To further validate IIS as a potential therapeutic target for treatment of ALS, manipulations of IIS in mammalian cells were investigated for neuroprotective activity. Genetic manipulations that increase the activity of the mammalian ortholog of daf-16, FOXO3, were found to be neuroprotective in a series of in vitro models of ALS toxicity.
    [Show full text]
  • A Novel Hypoxic Long Noncoding RNA KB-1980E6.3 Maintains Breast Cancer Stem Cell Stemness Via Interacting with IGF2BP1 to Facilitate C-Myc Mrna Stability
    Oncogene (2021) 40:1609–1627 https://doi.org/10.1038/s41388-020-01638-9 ARTICLE A novel hypoxic long noncoding RNA KB-1980E6.3 maintains breast cancer stem cell stemness via interacting with IGF2BP1 to facilitate c-Myc mRNA stability 1 2 3 4 1 1 1 1 1 Pengpeng Zhu ● Fang He ● Yixuan Hou ● Gang Tu ● Qiao Li ● Ting Jin ● Huan Zeng ● Yilu Qin ● Xueying Wan ● 1 1 5 1 Yina Qiao ● Yuxiang Qiu ● Yong Teng ● Manran Liu Received: 15 June 2020 / Revised: 13 November 2020 / Accepted: 18 December 2020 / Published online: 19 January 2021 © The Author(s), under exclusive licence to Springer Nature Limited 2021. This article is published with open access Abstract The hostile hypoxic microenvironment takes primary responsibility for the rapid expansion of breast cancer tumors. However, the underlying mechanism is not fully understood. Here, using RNA sequencing (RNA-seq) analysis, we identified a hypoxia-induced long noncoding RNA (lncRNA) KB-1980E6.3, which is aberrantly upregulated in clinical breast cancer tissues and closely correlated with poor prognosis of breast cancer patients. The enhanced lncRNA KB- 1980E6.3 facilitates breast cancer stem cells (BCSCs) self-renewal and tumorigenesis under hypoxic microenvironment both 1234567890();,: 1234567890();,: in vitro and in vivo. Mechanistically, lncRNA KB-1980E6.3 recruited insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) to form a lncRNA KB-1980E6.3/IGF2BP1/c-Myc signaling axis that retained the stability of c-Myc mRNA through increasing binding of IGF2BP1 with m6A-modified c-Myc coding region instability determinant (CRD) mRNA. In conclusion, we confirm that lncRNA KB-1980E6.3 maintains the stemness of BCSCs through lncRNA KB-1980E6.3/ IGF2BP1/c-Myc axis and suggest that disrupting this axis might provide a new therapeutic target for refractory hypoxic tumors.
    [Show full text]
  • Novel Regulators of the IGF System in Cancer
    biomolecules Review Novel Regulators of the IGF System in Cancer Caterina Mancarella 1, Andrea Morrione 2 and Katia Scotlandi 1,* 1 IRCCS Istituto Ortopedico Rizzoli, Laboratory of Experimental Oncology, 40136 Bologna, Italy; [email protected] 2 Department of Biology, Sbarro Institute for Cancer Research and Molecular Medicine and Center for Biotechnology, College of Science and Technology, Temple University, Philadelphia, PA 19122, USA; [email protected] * Correspondence: [email protected]; Tel.: +39-051-6366-760 Abstract: The insulin-like growth factor (IGF) system is a dynamic network of proteins, which includes cognate ligands, membrane receptors, ligand binding proteins and functional downstream effectors. It plays a critical role in regulating several important physiological processes including cell growth, metabolism and differentiation. Importantly, alterations in expression levels or activa- tion of components of the IGF network are implicated in many pathological conditions including diabetes, obesity and cancer initiation and progression. In this review we will initially cover some general aspects of IGF action and regulation in cancer and then focus in particular on the role of transcriptional regulators and novel interacting proteins, which functionally contribute in fine tuning IGF1R signaling in several cancer models. A deeper understanding of the biological relevance of this network of IGF1R modulators might provide novel therapeutic opportunities to block this system in neoplasia. Keywords: IGF system; cancer; transcriptional regulators; functional regulation; circular RNAs; IGF2BPs; ADAR; DDR1; E-cadherin; decorin Citation: Mancarella, C.; Morrione, A.; Scotlandi, K. Novel Regulators of the IGF System in Cancer. 1. Introduction Biomolecules 2021, 11, 273. https:// doi.org/10.3390/biom11020273 The insulin-like growth factor (IGF) system is a network of ligands, binding proteins and receptors regulating crucial physiological and pathological biological processes.
    [Show full text]
  • An Integrated Genome-Wide Multi-Omics Analysis of Gene Expression Dynamics in the Preimplantation Mouse Embryo
    bioRxiv preprint doi: https://doi.org/10.1101/495788; this version posted December 14, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. An integrated genome-wide multi-omics analysis of gene expression dynamics in the preimplantation mouse embryo Steffen Israel1,*, Mathias Ernst2,3,*, Olympia E. Psathaki4, Hannes C. A. Drexler1, Ellen Casser1, Yutaka Suzuki5, Wojciech Makalowski6, Michele Boiani1,†, Georg Fuellen2,†, Leila Taher2,3,† 1 Max-Planck-Institute for Molecular Biomedicine, Roentgenstr. 20, 48149 Muenster, Germany 2 Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock University Medical Center, Ernst-Heydemann Str. 8, 18057 Rostock, Germany 3 Bioinformatics, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstr. 5, 91058 Erlangen, Germany 4 University of Osnabrück, Center for Cellular Nanoanalytics Osnabrück (CellNanOs), Integrated Bioimaging Facility Osnabrück (iBiOs), Barbarastr. 11, 49076 Osnabrück, Germany 5 Department of Medical Genome Sciences, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba, 277-8562, Japan 6 Institute of Bioinformatics, Faculty of Medicine, University of Muenster, Niels Stensen Str. 14, 48149, Muenster, Germany. * These authors contributed equally † Corresponding authors: LT: [email protected]; GF: [email protected]; MB: [email protected]. Running Title: Multi-omics of the preimplantation mouse embryo Keywords: Preimplantation development, Proteome, Transcriptome, Model Organism 1 bioRxiv preprint doi: https://doi.org/10.1101/495788; this version posted December 14, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved.
    [Show full text]
  • IGF2BP3 Antibody
    Efficient Professional Protein and Antibody Platforms IGF2BP3 Antibody Basic information: Catalog No.: UPA62252 Source: Rabbit Size: 50ul/100ul Clonality: Polyclonal Concentration: 1mg/ml Isotype: Rabbit IgG Purification: Protein A affinity purified Useful Information: WB:1:1000-1:2000 Applications: IHC:1:50-1;200 FC:1:50-1:100 Reactivity: Human, Mouse, Rat Specificity: This antibody recognizes IGF2BP3 protein. Immunogen: Recombinant protein IGF-II mRNA-binding proteins (IMPs) bind RNA and influence RNA synthesis and metabolism. IMP-1, also known as coding region determinant-binding protein/insulin-like growth factor II mRNA-binding protein (CRD-BP) and VICKZ1; IMP-2 (IMP2, VICKZ2, p62); and IMP-3 (KOC1, VICKZ3) contain a unique combination of RNA recognition motifs and four hnRNP K homology domains. IMP-1 is abundant in embryonal tissues and is expressed in 81% of colon cancers, 73% of sarcomas and 58.5% of breast cancers. It recognizes Description: c-Myc, IGF-II and t mRNAs, and H19 RNA, and plays a major role in prolifera- tion of K-562 cells by an IGF-II-dependent mechanism. IMP-2 binds the 5' UTR of IGF-II mRNA and influences tumor cell growth, in which IMP-2 is as- sociated with apoptosis induced by tretinoin. IMP-3 knockdown by RNA in- terference decreases levels of IGF-II protein without affecting IGF-II, c-Myc, or β Actin mRNA and H19 RNA levels. IMP-3 is a marker for carcinomas and high-grade dysplastic lesions of pancreatic ductal epithelium. Uniprot: O00425(Human) Q9CPN8(Mouse) BiowMW: 63 kDa Buffer: 1*TBS (pH7.4), 1%BSA, 40%Glycerol.
    [Show full text]
  • Role of IGF2BP3 in Trophoblast Cell Invasion and Migration
    Citation: Cell Death and Disease (2014) 5, e1025; doi:10.1038/cddis.2013.545 OPEN & 2014 Macmillan Publishers Limited All rights reserved 2041-4889/14 www.nature.com/cddis Role of IGF2BP3 in trophoblast cell invasion and migration WLi1,2, D Liu2,3, W Chang4,XLu2,3, Y-L Wang2, H Wang2, C Zhu2, H-Y Lin2, Y Zhang5, J Zhou*,1 and H Wang*,2 The insulin-like growth factor-2 mRNA-binding protein 3 (IGF2BP3) is a member of a highly conserved protein family that is expressed specifically in placenta, testis and various cancers, but is hardly detectable in normal adult tissues. IGF2BP3 has important roles in RNA stabilization and translation, especially during early stages of both human and mouse embryogenesis. Placenta is an indispensable organ in mammalian reproduction that connects developing fetus to the uterine wall, and is responsible for nutrient uptake, waste elimination and gas exchange. Fetus development in the maternal uterine cavity depends on the specialized functional trophoblast. Whether IGF2BP3 plays a role in trophoblast differentiation during placental development has never been examined. The data obtained in this study revealed that IGF2BP3 was highly expressed in human placental villi during early pregnancy, especially in cytotrophoblast cells (CTBs) and trophoblast column, but a much lower level of IGF2BP3 was detected in the third trimester placental villi. Furthermore, the expression level of IGF2BP3 in pre-eclamptic (PE) placentas was significantly lower than the gestational age-matched normal placentas. The role of IGF2BP3 in human trophoblast differentiation was shown by in vitro cell invasion and migration assays and an ex vivo explant culture model.
    [Show full text]
  • IGF2BP3 Functions As a Potential Oncogene and Is a Crucial Target of Mir-34A in Gastric Carcinogenesis
    Zhou et al. Molecular Cancer (2017) 16:77 DOI 10.1186/s12943-017-0647-2 RESEARCH Open Access IGF2BP3 functions as a potential oncogene and is a crucial target of miR-34a in gastric carcinogenesis Yuhang Zhou1,2,3†, Tingting Huang1,2,3,4†, Ho Lam Siu1†, Chi Chun Wong2, Yujuan Dong2, Feng Wu1, Bin Zhang5, William K. K. Wu2,6, Alfred S. L. Cheng4,7, Jun Yu2,4,8, Ka Fai To1,2,3,4* and Wei Kang1,2,3,4* Abstract Background: Gastric cancer (GC) is one of the frequent causes of cancer-related death in eastern Asian population. IGF2BP2 lists in the top rank up-regulated genes in GC, but its functional role is unclear. Method: The expression of IGF2BP3 in GC cell lines and primary samples was examined by qRT-PCR and Western blot. The biological role of IGF2BP3 was revealed by a series of functional in vitro studies. Its regulation by microRNAs (miRNAs) was predicted by TargetScan and confirmed by luciferase assays and rescue experiments. Results: IGF2BP3 ranked the No.1 of the up-regulated genes by expression microarray analysis in GC cell lines. The expression level of IGF2BP3 was observed in GC tissues comparing with non-tumorous gastric epitheliums. The up-regulated IGF2BP3 expression was associated with poor disease specific survival. IGF2BP3 knockdown significantly inhibited cell proliferation and invasion. Apart from copy number gain, IGF2BP3 has been confirmed to be negatively regulated by tumor-suppressive miRNA, namely miR-34a. The expression of miR-34a showed negative correlation with IGF2BP3 mRNA expression in primary GC samples and more importantly, re-overexpression of IGF2BP3 rescued the inhibitory effect of miR-34a.
    [Show full text]
  • Supplementary Material Supplementary Figure1 A. EMSA
    Supplementary Material Supplementary Figure1 A. EMSA assay in THP-1 cells demonstrating increased protein binding to the probe with A allele compared to G allele. More DNA-protein complexes were shifted with the probe with allele A (lane 2) than allele G (lane 6); both unlabeled probes with A allele or G allele (24 pmol, 200X) can totally compete out the corresponding shifted band separately (lane 3 and lane 7); using 200X unlabeled probe with G allele can also compete out the shifted band formed with the probe with A allele (lane 4); unlabeled probe with A allele was also able to compete out the shifted band formed in the presence of probe with G allele (lane 8). B. The enhancer function of rs2069837 locus was also confirmed in THP-1 cells. Significantly increased luciferase expression was detected in both vectors with intronic sequence (A/G), with significantly higher expression in the construct with G allele than that with A allele (p< 0.05). C. The allele discrimination plot of 48 healthy subjects genotyped for rs2069837. D. No differences were observed between macrophages with AA and AG in the expressions of other genes interacting with rs2069837 based on DNase hypersensitivity correlations maps. E. Other epigenetic markers (H3K4me1 and H3K4me3) in monocytes also indicated the locus upstream GPNMB and the locus in the first intron of GPNMB are regulatory regions. GATA binding motif (72bp downstream of chr7:23248026 and 26bp upstream of chr7: 23288026) and MEF2 binding motif (149bp downstream of chr7:23248026 and 32bp upstream of chr7:23288026) close to these two loci are predicted, which were also demonstrated in ChIP-seq data of some cell lines (no ChIP-seq data of GATA and MEF2 in monocytes were available).
    [Show full text]
  • Igf2bp3 Maintains Maternal RNA Stability and Ensures Early Embryo Development in Zebrafish
    ARTICLE https://doi.org/10.1038/s42003-020-0827-2 OPEN Igf2bp3 maintains maternal RNA stability and ensures early embryo development in zebrafish Fan Ren 1,4, Qiaohong Lin 1,4, Gaorui Gong 1, Xian Du 2, Hong Dan 1, Wenying Qin 2, Ran Miao 1, ✉ Yang Xiong 1, Rui Xiao2, Xiaohui Li 1, Jian-Fang Gui 1,3 & Jie Mei 1 1234567890():,; Early embryogenesis relies on maternally inherited mRNAs. Although the mechanism of maternal mRNA degradation during maternal-to-zygotic transition (MZT) has been exten- sively studied in vertebrates, how the embryos maintain maternal mRNA stability remains unclear. Here, we identify Igf2bp3 as an important regulator of maternal mRNA stability in zebrafish. Depletion of maternal igf2bp3 destabilizes maternal mRNAs prior to MZT and leads to severe developmental defects, including abnormal cytoskeleton organization and cell division. However, the process of oogenesis and the expression levels of maternal mRNAs in unfertilized eggs are normal in maternal igf2bp3 mutants. Gene ontology analysis revealed that these functions are largely mediated by Igf2bp3-bound mRNAs. Indeed, Igf2bp3 depletion destabilizes while its overexpression enhances its targeting maternal mRNAs. Interestingly, igf2bp3 overexpression in wild-type embryos also causes a developmental delay. Altogether, these findings highlight an important function of Igf2bp3 in controlling early zebrafish embryogenesis by binding and regulating the stability of maternal mRNAs. 1 College of Fisheries, Huazhong Agricultural University, 430070 Wuhan, China. 2 Frontier Science Center for Immunology and Metabolism, Medical Research Institute, and Department of Hematology, Zhongnan Hospital of Wuhan University, Wuhan University, 430071 Wuhan, China. 3 State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, University of the Chinese Academy of ✉ Sciences, 430072 Wuhan, China.
    [Show full text]
  • IGF2BP3 Associates with Proliferative Phenotype and Prognostic Features in B-Cell Acute Lymphoblastic Leukemia
    cancers Article IGF2BP3 Associates with Proliferative Phenotype and Prognostic Features in B-Cell Acute Lymphoblastic Leukemia Artturi Mäkinen 1,2,* , Atte Nikkilä 1, Teppo Haapaniemi 2,3, Laura Oksa 1 , Juha Mehtonen 4 , Matti Vänskä 5 , Merja Heinäniemi 4, Timo Paavonen 2,6 and Olli Lohi 1,7 1 Tampere Center for Child, Adolescent and Maternal Health Research, Faculty of Medicine and Health Technology, Tampere University, 33520 Tampere, Finland; atte.nikkila@tuni.fi (A.N.); laura.oksa@tuni.fi (L.O.); olli.lohi@tuni.fi (O.L.) 2 Fimlab Laboratories, Department of Pathology, Tampere University Hospital, 33520 Tampere, Finland; teppo.haapaniemi@fimlab.fi (T.H.); timo.paavonen@tuni.fi (T.P.) 3 Department of Biological and Environmental Sciences, University of Jyväskylä, 40014 Jyväskylä, Finland 4 Institute of Biomedicine, School of Medicine, University of Eastern Finland, 70211 Kuopio, Finland; juha.mehtonen@uef.fi (J.M.); merja.heinaniemi@uef.fi (M.H.) 5 Department of Internal Medicine, Tampere University Hospital, 33520 Tampere, Finland; matti.vanska@pshp.fi 6 Department of Pathology, Faculty of Medicine and Health Technology, Tampere University, 33520 Tampere, Finland 7 Tays Cancer Centre, Tampere University Hospital, 33520 Tampere, Finland * Correspondence: artturi.makinen@tuni.fi Simple Summary: Although the prognosis of acute lymphoblastic leukemia (ALL) has improved Citation: Mäkinen, A.; Nikkilä, A.; significantly during the past decades, ALL remains a major cause of pediatric cancer mortality, and Haapaniemi, T.; Oksa, L.; Mehtonen, more accurate risk-stratification is required. We investigated IGF2BP3, which has previously been J.; Vänskä, M.; Heinäniemi, M.; associated with aggressive cancers, and found high and subtype-specific expression of IGF2BP3 Paavonen, T.; Lohi, O.
    [Show full text]
  • The Role of the Oncofetal IGF2 Mrna-Binding Protein 3 (IGF2BP3)
    G Model YSCBI-1158; No. of Pages 10 ARTICLE IN PRESS Seminars in Cancer Biology xxx (2014) xxx–xxx Contents lists available at ScienceDirect Seminars in Cancer Biology j ournal homepage: www.elsevier.com/locate/semcancer Review The role of the oncofetal IGF2 mRNA-binding protein 3 (IGF2BP3) in cancer a a,b a a,b,∗ Marcell Lederer , Nadine Bley , Christian Schleifer , Stefan Hüttelmaier a Division of Molecular Cell Biology, Institute of Molecular Medicine, Martin Luther University Halle-Wittenberg, Heinrich-Damerow-Strasse 1, 06120 Halle, Germany b Core Facility Imaging (CFI) of the Medical Faculty, Institute of Molecular Medicine, Martin Luther University Halle-Wittenberg, Heinrich-Damerow-Strasse 1, 06120 Halle, Germany a r t i c l e i n f o a b s t r a c t Keywords: The post-transcriptional control of gene expression mediated by RNA-binding proteins (RBPs), long non- IGF2 mRNA-binding protein coding RNAs (lncRNAs) as well as miRNAs is essential to determine tumor cell fate and thus is a major IMP determinant in cancerogenesis. The IGF2 mRNA binding protein family (IGF2BPs) comprises three RBPs. VICKZ Two members of the family, IGF2BP1 and IGF2BP3, are bona fide oncofetal proteins, which are de novo CRD-BP synthesized in various human cancers. In vitro studies revealed that IGF2BPs serve as post-transcriptional IGF2BP fine-tuners modulating the expression of genes implicated in the control of tumor cell proliferation, sur- vival, chemo-resistance and metastasis. Consistently, the expression of both IGF2BP family members was reported to correlate with an overall poor prognosis and metastasis in various human cancers.
    [Show full text]
  • Combinatorial Recognition of Clustered RNA Elements by a Multidomain RNA-Binding Protein, IMP3
    bioRxiv preprint doi: https://doi.org/10.1101/398008; this version posted August 22, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Combinatorial recognition of clustered RNA elements by a multidomain RNA-binding protein, IMP3 Tim Schneider1,6, Lee-Hsueh Hung1,6, Masood Aziz2, Anna Wilmen1, Stephanie Thaum1, Jacqueline Wagner2, Robert Janowski3, Simon Müller4, Stefan Hüttelmaier4, Dierk Niessing3,5, Michael Sattler2*, Andreas Schlundt2*, and Albrecht Bindereif1* 1 Institute of Biochemistry, University of Giessen, 35392 Giessen, Germany 2 Center for Integrated Protein Science Munich (CIPSM) at Department of Chemistry, Technical University of Munich (TUM), 85747 Garching, Germany 3 Institute of Structural Biology, Helmholtz-Zentrum München, 85764 Neuherberg, Germany 4 Institute of Molecular Medicine, Section for Molecular Cell Biology, Faculty of Medicine, Martin Luther University Halle-Wittenberg, 06120 Halle, Germany 5 Institute of Pharmaceutical Biotechnology, Ulm University, 89081 Ulm, Germany 6 These authors contributed equally to this work. * Correspondence should be addressed to: M.S. ([email protected]), A.S. ([email protected]), or A.B. ([email protected]). 1 bioRxiv preprint doi: https://doi.org/10.1101/398008; this version posted August 22, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract How multidomain RNA-binding proteins recognize their specific target sequences, based on a combinatorial code, represents a fundamental unsolved question and has not been studied systematically so far.
    [Show full text]