Journal: Translational Cancer Research

Article Title: Aspirin inhibits proliferation and metastasis of canine mammary gland tumor cells through Wnt signaling axis

doi: 10.21037/tcr-20-3172

Figure Lengend Snippet: Aspirin inhibits the canonical Wnt/β-catenin pathway in CMGTs. Western blotting showed that Wnt3a, MMP-2, MMP-9, VEGF, p-GSK-3β, PPAR-δ and c-myc were decreased while β-catenin and E-cadherin were increased in CHMp (A) and CHMm (B) cells treated with high concentrations of aspirin (2.5, 5, or 10 mM). Results were expressed as mean ± SD of three independent experiments (n=3). ***, P<0.001 when compared to untreated group.

Article Snippet: Then, 0.1% Triton-X100 1 mL at room temperature was added to these cells for 15 minutes, and incubated with antibodies (β-catenin, p-GSK3β from Bioss, Beijing, China) for 12 h at 4 °C.

Techniques: Western Blot

Journal: Translational Cancer Research

Article Title: Aspirin inhibits proliferation and metastasis of canine mammary gland tumor cells through Wnt signaling axis

doi: 10.21037/tcr-20-3172

Figure Lengend Snippet: Aspirin inhibited the expression and nuclear translocation of β-catenin and p-GSK-3β in CMGTs. CHMp (A) and CHMm (B) cells were seeded onto glass for 15 h and then treated with varying concentrations of Aspirin for 36 h. β-catenin and p-GSK-3β (red) localization was visualized by immunofluorescence followed by confocal imaging. DAPI was used to stain the nuclear (blue). Images are representative of three-independent experiments. Results were expressed as mean ± SD of three independent experiments (n=3).

Article Snippet: Then, 0.1% Triton-X100 1 mL at room temperature was added to these cells for 15 minutes, and incubated with antibodies (β-catenin, p-GSK3β from Bioss, Beijing, China) for 12 h at 4 °C.

Techniques: Expressing, Translocation Assay, Immunofluorescence, Imaging, Staining

Journal: Translational Cancer Research

Article Title: Aspirin inhibits proliferation and metastasis of canine mammary gland tumor cells through Wnt signaling axis

doi: 10.21037/tcr-20-3172

Figure Lengend Snippet: Wnt3a attenuated aspirin-induced inhibition of Wnt/β-catenin signaling and migration and invasion of CMGTs. Wound healing (A,B,C) and Transwell invasion assays (B,C,D,E,F) showed that treatment with Wnt3a (0.1 µg/mL) promoted migration and invasion of CMGTs compared to the untreated group or treatment with only aspirin (5 mM) (0.1% crystal violet staining, magnification, 200×). (G,H,I) Wnt3a elevated p-GSK-3β, β-catenin, MMP-2, MMP-9 and CyclinD1, and it decreased E-cadherin. Results were expressed as mean ± SD of three independent experiments (n=3). *, P<0.05; **, P<0.01; ***, P<0.001 when compared to untreated group; ### , P<0.001 compared to Aspirin group.

Article Snippet: Then, 0.1% Triton-X100 1 mL at room temperature was added to these cells for 15 minutes, and incubated with antibodies (β-catenin, p-GSK3β from Bioss, Beijing, China) for 12 h at 4 °C.

Techniques: Inhibition, Migration, Staining

Journal: Translational Cancer Research

Article Title: Aspirin inhibits proliferation and metastasis of canine mammary gland tumor cells through Wnt signaling axis

doi: 10.21037/tcr-20-3172

Figure Lengend Snippet: FH535 inhibits cell migration, and invasion in CMGTs. (A,B) Wound healing (A) and Transwell invasion assay (B) showed that treatment with the combination of FH535 and Aspirin inhibited migration and invasion of CMGTs compared with untreated group, but as the same effect of treatment with only aspirin (5 mM) or only FH535 (10 uM) (0.1% crystal violet staining, magnification, 200×). (C,D) FH535 elevated E-cadherin, and it decreased p-GSK-3β, β-catenin, MMP-2, MMP-9 and CyclinD1.There was no significant difference between combined drug use and single drug use. Results were expressed as mean ± SD of three independent experiments (n=3). ***, P<0.001 when compared to untreated group.

Article Snippet: Then, 0.1% Triton-X100 1 mL at room temperature was added to these cells for 15 minutes, and incubated with antibodies (β-catenin, p-GSK3β from Bioss, Beijing, China) for 12 h at 4 °C.

Techniques: Migration, Transwell Invasion Assay, Staining