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Cancer reports (Hoboken, N.J.)
10, 2019;2 (5): e1204.

Tumorigenic effects of TLX overexpression in HEK 293T cells.

Toshima Z Parris, Dzeneta Vizlin-Hodzic, Susanne Salmela, Keiko Funa
Author Information
  1. Toshima Z Parris: Department of Oncology, Institute of Clinical Sciences, Sahlgrenska Cancer Center, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden. ORCID
  2. Dzeneta Vizlin-Hodzic: Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
  3. Susanne Salmela: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Cancer Center, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
  4. Keiko Funa: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Cancer Center, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
PMID: 32721119 DOI: 10.1002/cnr2.1204

Abstract

BACKGROUND: The human orphan receptor TLX (NR2E1) is a key regulator of neurogenesis, adult stem cell maintenance, and tumorigenesis. However, little is known about the genetic and transcriptomic events that occur following TLX overexpression in human cell lines.
AIMS: Here, we used cytogenetics and RNA sequencing to investigate the effect of TLX overexpression with an inducible vector system in the HEK 293T cell line.
METHODS AND RESULTS: Conventional spectral karyotyping was used to identify chromosomal abnormalities, followed by fluorescence in situ hybridization (FISH) analysis on chromosome spreads to assess TLX DNA copy number. Illumina paired-end whole transcriptome sequencing was then performed to characterize recurrent genetic variants (single nucleotide polymorphisms (SNPs) and indels), expressed gene fusions, and gene expression profiles. Lastly, flow cytometry was used to analyze cell cycle distribution. Intriguingly, we show that upon transfection with a vector containing the human TLX gene (eGFP-hTLX), an isochromosome forms on the long arm of chromosome 6, thereby resulting in DNA gain of the TLX locus (6q21) and upregulation of TLX. Induction of the eGFP-hTLX vector further increased TLX expression levels, leading to G0-G1 cell cycle arrest, genetic aberrations, modulation of gene expression patterns, and crosstalk with other nuclear receptors (AR, ESR1, ESR2, NR1H4, and NR3C2). We identified a 49-gene signature associated with central nervous system (CNS) development and carcinogenesis, in addition to potentially cancer-driving gene fusions (LARP1-CNOT8 and NSL1-ZDBF2) and deleterious genetic variants (frameshift insertions in the CTSH, DBF4, POSTN, and WDR78 genes).
CONCLUSION: Taken together, these findings illustrate that TLX may play a pivotal role in tumorigenesis via genomic instability and perturbation of cancer-related processes.

Keywords

NR2E1 TLX cancer genome instability nuclear receptors

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MeSH Term

Cell Proliferation
Cell Transformation, Neoplastic
Frameshift Mutation
G1 Phase Cell Cycle Checkpoints
Gene Expression Regulation, Neoplastic
Genomic Instability
HEK293 Cells
Humans
Orphan Nuclear Receptors
RNA-Seq
Up-Regulation

Chemicals

NR2E1 protein, human
Orphan Nuclear Receptors
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 1

Word Cloud

Created with Highcharts 10.0.0TLXcellgenegenetichumanoverexpressionusedvectorexpressionNR2E1tumorigenesissequencingsystemHEK293TchromosomeDNAvariantsfusionscycleeGFP-hTLXnuclearreceptorsinstabilityBACKGROUND:orphanreceptorkeyregulatorneurogenesisadultstemmaintenanceHoweverlittleknowntranscriptomiceventsoccurfollowinglinesAIMS:cytogeneticsRNAinvestigateeffectinduciblelineMETHODSANDRESULTS:ConventionalspectralkaryotypingidentifychromosomalabnormalitiesfollowedfluorescencesituhybridizationFISHanalysisspreadsassesscopynumberIlluminapaired-endwholetranscriptomeperformedcharacterizerecurrentsinglenucleotidepolymorphismsSNPsindelsexpressedprofilesLastlyflowcytometryanalyzedistributionIntriguinglyshowupontransfectioncontainingisochromosomeformslongarm6therebyresultinggainlocus6q21upregulationInductionincreasedlevelsleadingG0-G1arrestaberrationsmodulationpatternscrosstalkARESR1ESR2NR1H4NR3C2identified49-genesignatureassociatedcentralnervousCNSdevelopmentcarcinogenesisadditionpotentiallycancer-drivingLARP1-CNOT8NSL1-ZDBF2deleteriousframeshiftinsertionsCTSHDBF4POSTNWDR78genesCONCLUSION:Takentogetherfindingsillustratemayplaypivotalroleviagenomicperturbationcancer-relatedprocessesTumorigeniceffectscellscancergenome

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