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, supplemented with 1% (v/v) antibiotic (Streptomycin and Penicillin, Gibco?) and 10% (v/v) fetal bovine serum (FBS, Dutscher, Bhas 42 cells were obtained from Harlan Laboratories, p.10
, Cell transformation and cell survival assays The CTA in promotion condition, originally proposed by the OECD
, Supplementary Table A2). Amplification was performed following the manufacturer
, TUBA (alpha-Tubulin), and GAPDH (glyat 4°C. Supernatants were collected and stored at ?80°C. Protein concentration was measured in each sample and corresponding cell medium supernatant, using bovine serum albumin as a standard, ACTB (beta-Actin)
, ?g) of whole cell homogenates or 20 ?l of corresponding cell medium supernatants were resolved in 4-20% SDS PAGE (CriterionTM Tris-Glycine eXtended Stain-Free Protein Gel, BioRad) and transferred onto polyvinylidene difluoride (PVDF) membranes (BioRad), using semi-dry transfert (BioRad). and Ninein interacting protein (AUNIP, 500), Galectin-7, vol.1, p.1000, 206435.
, Mini Chromosome Maintenance deficient 10 (MCM10, vol.1, p.1000
Gene clustering was visualized with Treeview. An uncentred correlation was used as a measure of similarity and complete linkage was used for the clustering method. (B) Dose effect of NM-203 (1-5 ?g/cm 2 ). Supervised hierarchical clustering was performed with Gene Cluster 3.0 software and visualized by Treeview on the 285 probes obtained after filtration and application of the ANOVA statistical test, Min-U-Sil 5® (20 ?g/cm 2 ), vol.1, pp.0-203 ,
, TPA, 12-O-tetradecanoylphorbol-13-acetate
,
5000) or HRP-conjugated anti-rabbit IgG (1: 2000) in PBST containing 5% (w/v) non-fat milk. Finally, membranes were washed 3 times with PBST before chemiluminescence detection (Clarity ? Western ECL Substrate, BioRad) using ChemiDocTM (ChemiDoc ? System, BioRad), For interpretation of the references to color in HRP-conjugated anti-goat IgG ,
,
, An unpaired t-test was used for two by two comparison (treatments vs. control (H 2 O or DMSO)). Data are given as a mean ± standard deviation (mean ± SD)
, Gene set enrichment analysis. (A) GSEA analysis using the filtered gene expression profiles of Bhas 42 cells 48 h after treatment with NM-203 (5 ?g/cm 2 ), Min-U-Sil 5® (20 ?g/cm 2 ) and TPA
, List of the 12 common signatures. (C) GSEA revealed an enrichment in gene signatures related to cell cycles and cancer, ?g/ml) over 48 h. Venn diagram of the top 50 signatures. (B)
, Chemico-Biological Interactions, vol.315, p.108900, 2020.
, Genes significantly highly regulated by NM-203 (5 ?g/cm 2 ), Min-U
Of the 309 genes altered by treatments, a large majority (83%) were affected by NM-203. Among these, 142 (46%) were affected solely by NM-203 treatment and 23 (7%) and 29 genes (9%) were deregulated by Min-U-Sil 5® and TPA, respectively. Interestingly, 21 common genes were regulated significantly by NM-203 (5 ?g/cm 2 ), Min-U-Sil 5® and TPA (p < 0.005, FC > 4). Twelve genes, Sil 5® and TPA were compared (p < 0.005 and FC > 4) (Fig. 4A) ,
Transforming silica nanoparticles NM-200 and NM-202 induce similar changes at the RNA level of the selected genes We then assessed whether Bhas 42 cell exposure to other SAS, namely NM-200, NM 201 and NM202, led to cell transformation and involved any changes at the RNA level of the selected genes, NM-200 of AUNIP, IL1RL1, MCM10, PRL2C1, PRL2C3 and PRL2C5, and a decrease for LGALS7 ,
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,
U-Sil 5 Ò , and with DMSO as the vehicle for TPA. A logarithmic transformation was applied to the data ,
, List of significantly regulated genes after NM-203 (5 µg/cm 2 ) treatment (unpaired T-test with p<0.05, FC>2). FC: Fold Change
, List of significantly regulated genes after Min-U-Sil 5 Ò, p.20
, µg/cm 2 ) treatment (unpaired T-test with p<0.05, FC>2). FC: Fold Change
, List of significantly regulated genes after TPA (0.05 µg/ml) treatment (unpaired T-test with p<0.05, FC>2). FC: Fold Change
, List of significantly regulated genes after NM-203 (2 µg/cm 2 ) treatment (unpaired T-test with p<0.05, FC>2). FC: Fold Change
, List of significantly regulated genes after NM-203 (5 µg/cm 2 ) treatment (unpaired T-test with p<0.005, FC>4). FC: Fold Change
, List of significantly regulated genes after Min-U-Sil 5 Ò, p.20
, µg/cm 2 ) treatment (unpaired T-test with p<0.005, FC>4). FC: Fold Change
, List of significantly regulated genes after TPA (0.05 µg/ml) treatment (unpaired T-test with p<0.005, FC>4). FC: Fold Change
, List of significantly regulated genes after NM-203 (2 µg/cm 2 ) treatment (unpaired T-test with p<0.005, FC>4). FC: Fold Change
, The top common canonical pathways are highlighted by IPA. Ctrl: H2O; TPA: 12-Otetradecanoylphorbol-13-acetate; DE: Diatomaceous earth; DMSO: Dimethyl sulfoxide; FC: Fold Change