Article Figures & Data
Figures
- Figure 1
The miR-155 showed a negative relationship with adipocyte differentiation of adipose mesenchymal stem cells (A-MSCs) in gastric cancer (GC) patients. (A) Relationship between miR-155 and GC survival from the tumor database. (B) Relative levels of miR-155 in serum from GC patients and healthy donors using RT-PCR (N = 150). (C) Oil Red O staining analysis of the adipo-differentiation capacity of A-MSCs from GC patients and healthy donors (N = 5, scale bar = 100 μm). (D, E) Western blot analysis of C/EBPβ expressions in A-MSCs in GC patients (N = 5). (F) Relative levels of miR-155 in A-MSCs from GC patients and healthy donors (N = 5). *P < 0.05; **P < 0.01; ***P < 0.001.
- Figure 2
Gastric cancer exosomes inhibited adipocyte differentiation of adipose mesenchymal stem cells (A-MSCs). (A) Oil Red O staining was performed to visualize lipid droplet accumulation in A-MSCs treated with GES-1-Exo or SGC7901-Exo. (N = 3, scale bar = 100 μm). (B) RT-PCR analysis of adipogenic-specific genes (C/EBPβ, C/EBPα, and PPARγ were normalized to β-actin) in the groups described above (N = 3). (C) Western blot analysis of C/EBPβ, C/EBPα, and PPARγ in the groups described above (N = 3). (D) Oil Red O staining was performed to detect lipid droplet accumulation in A-MSCs treated with GES-1-Exo or MGC803-Exo. (N = 3, scale bar = 100 μm). (E) RT-PCR analysis of adipogenic-specific genes and (F) western blot analysis of the expression of C/EBPβ, C/EBPα, and PPARγ in the groups described above (N = 3). *P < 0.05; **P < 0.01.
- Figure 3
Gastric cancer cell-derived exosomes promoted browning of white adipose tissue. (A) Representative images of MitoTracker staining (red labeling) with BODIPY staining for lipid droplets (green labeling) (N = 3, scale bar = 10 μm). (B) Immunofluorescence assay for UCP1 (red labeling) with BODIPY staining for lipid droplets (green labeling) (N = 3, scale bar = 10 μm). (C) Western blot analysis of UCP1 expressions in adipose mesenchymal stem cells (A-MSCs) treated with GES-1-Exo and SGC7901-Exo. (D) Western blot analysis of UCP1 expression in A-MSCs treated with GES-1-Exo and MGC803-Exo (N = 3). (E) Relative ATP levels of A-MSCs treated with GES-1-Exo and SGC7901-Exo (N = 3). (F) Relative ATP levels of A-MSCs treated with GES-1-Exo and MGC803-Exo (N = 3). *P < 0.05; **P < 0.01.
- Figure 4
Gastric cancer-exosome-miR-155 targeting C/EBPβ in adipose mesenchymal stem cells (A-MSCs). (A) Predicted binding sites for miR-155 in the 3′-UTR of C/EBPβ mRNA. (B) Direct recognition of C/EBPβ by miR-155. Firefly luciferase reporters containing either the wild-type (WT) or mutant (MUT) C/EBPβ 3′-UTR sequence, miR-155 mimics, miR-155 inhibitors and the corresponding normal control were co-transfected into A-MSCs. The relative luciferase levels were detected (N = 3). (C–D) RT-PCR analysis of miR-155 levels in A-MSCs transfected with normal control mimics (Mi NC), miR-155 mimics (Mi miR-155), normal control inhibitors (In NC), and miR-155 inhibitors (In miR-155) (N = 3). (E) RT-PCR analysis of C/EBPβ mRNA levels in the groups described above (N = 3). (F–G) C/EBPβ expression in the groups described above (N = 3). (H) Schematic description of the experimental design. Isolation of exosomes after inhibiting miR-155 levels in SGC7901 cells and normal controls and adding to A-MSCs. (I) RT-PCR assay of miR-155 levels in SGC exo In.miR-155 or SGC exo In.NC (N = 3). (J) RT-PCR assay of miR-155 levels in A-MSCs treated with SGC exo In.miR-155 or SGC exo In.NC (N = 3). (K) C/EBPβ mRNA levels in A-MSCs pretreated with different exosomes were detected by RT-PCR (N = 3). (L–M) C/EBPβ, C/EBPα, PPARγ, and UCP1 expressions in A-MSCs treated with SGC exo In.miR-155 and SGC exo In.NC analyzed by western blotting (N = 3). **P < 0.01.
- Figure 5
Influence of exosomal miR-155 in gastric cancer on loss of adipose tissue in vivo. (A) Flow chart depicting the in vivo experimental design. (B) Weight change of the mice (N = 3). (C) Morphology of the inguinal adipose tissue after the mice were injected with PKH67-labeled SGC7901 exosomes (scale bar = 100 μm). (D–E) Images of inguinal adipose tissue and analysis of the length (N = 6). (F) Expression of UCP1 in inguinal adipose tissue was identified by immunohistochemistry (N = 3, scale bar = 100 μm). (G) Representative images from hematoxylin and eosin (H&E)-stained inguinal adipose tissue (N = 3, scale bar = 100 μm). *P < 0.05; **P < 0.01.
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