biostudies-arrayexpressAnda GligaHomo sapiensRnBeads packagehttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-6331Human bronchial lung cells (BEAS-2B) were treated for 6 weeks with low doses (1 µg/mL) of Ag nanoparticles (10 nm citrate-coated). At the end of the exposure control and treated samples were submitted for DNA methylation analysis (Illumina 450k). The overall goal of this experiment was to evaluate potential epigenetic changes associated with low-dose, long-term exposure to Ag nanoparticles.biostudies-arrayexpressLabeling - Illumina’s protocol Infinium HDNucleic Acid Extraction - At the end of exposure week 6, DNA was extracted using the Invisorb Spin Tissue Mini Kit (Stratec) in accordance with manufacturer’s instructions.Scaning - Illumina’s protocol Infinium HDHybridization - Illumina’s protocol Infinium HDGrowth Protocol - BEAS-2B cells (European Collection of Cell Cultures) were cultured in bronchial epithelial cell growth medium (BEGM, Lonza) supplemented with BEGM bullet-kit (Lonza: recombinant epidermal growth factor (EGF), hydrocortisone, insulin, bovine pituitary extract, GA-1000 (Gentamicin Sulfate and Amphotericin-B), retinoic acid, transferrin, triiodothyronine and epinephrine). Cells were cultured in flasks and plates pre-coated with: 0.01 mg/mL fibronectin, 0.03 mg/mL bovine collagen type I, 0.01 mg/mL bovine serum albumin and 0.2% penicillin-streptomycin in BEGM additive free medium for 1-2 h prior to the seeding. Cells were maintained in a humidified atmosphere at 37˚C, 5% CO2 and sub-cultured at 80% confluency.Sample Treatment - For the long-term exposure, BEAS-2B were seeded in 6 well-plates (5000 cells/cm2, 2 mL cell medium per well) and allowed to attach for approx. 2 h. Thereafter, cells were exposed to 1 µg/mL Ag10 or Ag75 (approx. 0.2 µg/cm2) in triplicates. Cells were split (trypsinization), counted, reseeded twice a week (5000 cells/ cm2, 2 mL cell medium per well) and re-exposed to the Ag nanoparticles.MIAME ScoreRaw DataOrganizationAssays and DataMAGE-TAB FilesArray DesignsData Transformation - methylumi-noob for background correction and dasen normalizationMetabolomicsUnknownTranscriptomicsGenomicsProteomicsDespite a considerable focus on the adverse effects of silver nanoparticles (AgNPs) in recent years, studies on the potential long-term effects of AgNPs are scarce. The aim of this study was to explore the effects of AgNPs following repeated low-dose, long-term exposure of human bronchial epithelial cells. To this end, the human BEAS-2B cell line was exposed to 1 µg/mL AgNPs (10 nm) for 6 weeks followed by RNA-sequencing (RNA-Seq) as well as genome-wide DNA methylation analysis. The transcriptomics analysis showed that a substantial number of genes (1717) were differentially expressed following AgNP exposure whereas only marginal effects on DNA methylation were observed. Downstream analysis of the transcriptomics data identified several affected pathways including the 'fibrosis' and 'epithelial-mesenchymal transition' (EMT) pathway. Subsequently, functional validation studies were performed using AgNPs of two different sizes (10 nm and 75 nm). Both NPs increased collagen deposition, indicative of fibrosis, and induced EMT, as evidenced by an increased invasion index, anchorage independent cell growth, as well as cadherin switching. In conclusion, using a combination of RNA-Seq and functional assays, our study revealed that repeated low-dose, long-term exposure of human BEAS-2B cells to AgNPs is pro-fibrotic, induces EMT and cell transformation.methylation profiling by arrayHomo sapiensRNA-sequencing reveals long-term effects of silver nanoparticles on human lung cells.Gliga AR, Di Bucchianico S, Lindvall J, Fadeel B, Karlsson HL.Anda GligafalseDNA methylation profiling of BEAS-2B cells exposed to low doses of Ag nanoparticles for 6 weeksHuman bronchial lung cells (BEAS-2B) were treated for 6 weeks with low doses (1 µg/mL) of Ag nanoparticles (10 nm citrate-coated). At the end of the exposure control and treated samples were submitted for DNA methylation analysis (Illumina 450k). The overall goal of this experiment was to evaluate potential epigenetic changes associated with low-dose, long-term exposure to Ag nanoparticles.2018-04-30T00:00:00Z2026-03-13T17:22:36.801Z2022-02-04T04:13:26.066ZE-MTAB-633129703973E-MTAB-6321EFO_0002944EFO_0003814EFO_0003813EFO_0003789EFO_0002759EFO_0003816EFO_0003969EFO_000381510.1038/s41598-018-25085-5