Additional file 1 of TiO2 nanoparticles promote tumor metastasis by eliciting pro-metastatic extracellular vesicles
Creators: name: Mu, Xupeng
name: Hu, Kebang
name: Wei, Anhui
name: Bai, Jinping
name: Feng, Li
name: Jiang, Jinlan
Publisher: name: figshare
Year: 2023
Subjects: Space Science
Medicine
Cell Biology
Molecular Biology
Pharmacology
Biological Sciences not elsewhere classified
Developmental Biology
Cancer

description: Additional file 1: Figure S1. TiO2 inorganic NPs promoted tumor metastasis in vivo. a Body weight of mice treated with PBS or TiO2 during the period of 21 days post-tumor challenge. b Tumor weight of mice treated with PBS or TiO2 during the period of 21 days post-tumor challenge. Data are presented as mean value ± SD (n = 6). Statistical analysis was measured by one-way ANOVA. Figure S2. TEM images of TiO2 NPs with different sizes. a 30 nm, b 60 nm. Scale bars = 100 nm. c 100 nm. Scale bars = 200 nm. Figure S3. Tumor volume (a) and tumor weight (b) of mice treated with PBS-EVs or TiO2-EVs during the period of 21 days post-tumor challenge. Data are presented as mean value ± SD (n = 6). Statistical analysis was measured by one-way ANOVA. Figure S4. The effects of PBS-EVs or TiO2-EVs on tumor migration ability. Wound healing assay (a) and transwell assay (b) were used to detect the effects of PBS-EVs or TiO2-EVs on tumor migration ability. Scale bars = 50 μm. Data are presented as mean value ± SD (n = 3). Statistical analysis was measured by one-way ANOVA. Figure S5. Immunofluorescence analysis of the expression of Von Willebrand factor (VWF) and platelet endothelial cell adhesion molecule-1 (CD31) in separated HUVECs. Scale bars = 20 μm. Nuclei, Blue (DAPI); CD31 or VWF, Red. Figure S6. Cell-Tracker Green labeled MCF-7 cells (a) and Cell-Tracker Red labeled A549 cells (b) migrated through HUVECs layers were observed under fluorescent microscopy following exposure of HUVECs to EVs for 1 h. Scale bars = 50 μm. c Experimental scheme for the migration assay. Figure S7. The effects of PBS-EVs or TiO2-EVs on HUVECs migration ability. Wound healing assay (a) and transwell assay (b) were used to detect the effects of PBS-EVs or TiO2-EVs on HUVECs migration ability. Scale bars = 50 μm. Data are presented as mean value ± SD (n = 3). Statistical analysis was measured by one-way ANOVA. Figure S8. a Cluster diagram of differential expression miRNA. Each row represented one miRNA, and each column represented one sample. The red dots and green dots represented the upregulated and downregulated miRNAs, respectively b Scatter Plot of the DE-miRNAs. The black dots represent miRNAs that are not differentially expressed between PBS-EVs and TiO2-EVs, and the red dots and green dots represent the upregulated and downregulated miRNAs, respectively. Figure S9. GO functions for the potential target genes of the top upregulated miRNAs were performed, including a Biological process (BP), b Cellular component (CC), c Molecular function (MF). d KEGG signaling pathway enrichment analysis were performed for significant differences in cancer related genes between PBS-EVs and TiO2-EVs. Figure S10. a Prediction of the binding sites of miR-301a-3p and 3′-UTR of S1PR1. b Construction of a reporter gene plasmid containing wild-type (wt) or mutant (mt) 3′-UTR of S1PR1 binding sites. Figure S11. The relative luciferase activity of wild-type (wt) and mutant-type (mt) 3′-UTR of S1PR1 was determined in HUVECs. miR-301a-3p inhibited the luciferase activity of reporter containing wild-type but not mutant 3′-UTR of S1PR1. Data are shown as mean ± SD (n = 3). Statistical analysis was measured by one-way ANOVA. Figure S12. RT-PCR analysis of miR-301a-3p expression in EVs derived from MDA-MB-231 cells which were treated with a miR-301a-3p mimic. Data are shown as mean ± SD (n = 3). Statistical analysis was measured by one-way ANOVA. Figure S13. Transwell assay were used to detect the effects of EVs-con or EVs-miR on HUVECs migration ability. Scale bars = 50 μm. Data are presented as mean value ± SD (n = 3). Statistical analysis was measured by one-way ANOVA. Figure S14. HE staining images of the major organs of mice from different treatment groups. The results indicated that the EVs had no obvious toxic effects on the organs of mice. Scale bar = 50 µm. Figure S15. The membrane images for all Western blotting reported in the main figures of this study. Table S1. The primers used in the mRNA/miRNA expression analysis experiment.