Fig. 3: AMPC inhibits the oncogenicity of ER+HER2+ MC cells. | Cell Death & Disease

Fig. 3: AMPC inhibits the oncogenicity of ER+HER2+ MC cells.

From: Inhibition of TFF3 synergizes with c-MET inhibitors to decrease the CSC-like phenotype and metastatic burden in ER+HER2+ mammary carcinoma

Fig. 3

A BrdU incorporation assay in MDA-MB-361 and BT474 cells treated with 0, 2.5, 5, or 10 µM AMPC for 3 days. Data are expressed as mean ± SD (n = 3). Statistical significance is indicated as *P < 0.05, **P < 0.01, and ***P < 0.001. B CASPASE 3/7 activities were determined after 3 days of AMPC (0, 2.5, 5, or 10 µM) treatment. Data are expressed as mean ± SD (n = 3). Statistical significance is indicated as *P < 0.05, **P < 0.01, and ***P < 0.001. C Apoptotic cell death of MDA-MB-361 and BT474 cells was determined after 2 days of AMPC treatment (0, 2.5, 5, or 10 µM) by Annexin-V/PI. The percentages of early apoptotic and late apoptotic are shown. Data are expressed as mean ± SD (n = 3). Statistical significance is indicated as *P < 0.05, **P < 0.01, and ***P < 0.001. D Foci formation was conducted with MDA-MB-361 and BT474 cells treated with 0, 2.5, 5, or 10 µM AMPC in monolayer culture at low cell density for 14 days. Foci were stained by crystal violet. E Microscopic visualization of colonies stained with the LIVE/DEAD™ Cell Imaging Kit after 12 days of AMPC treatment (0, 2.5, 5, or 10 µM) of MDA-MB-361 and BT474 cells cultured in medium containing 2% FBS and 4% Matrigel after 3 days of pre-culture. The bright-field images are shown above, and the merged images of live (green) and dead (red) colonies are shown below. Scale bars, 100 μm. F Spheroid formation assay. MDA-MB-361 and BT474 cells were seeded in ultralow attachment plates and cultured in spheroid growth media for 12 days with AMPC treatment (0, 2.5, 5, or 10 µM). Scale bar, 100 μm. G ALDEFLUOR activity was measured after 3 days of AMPC treatment (0, 2.5, 5, or 10 µM). MDA-MB-361 and BT474 cells were then harvested and incubated with ALDEFLUOR substrate to define the ALDH1-positive population. DEAB was used to establish the baseline fluorescence. The percentage of ALDH1-positive cells (in the pink box) was plotted by flow cytometry. Data are expressed as mean ± SD (n = 3). Statistical significance is indicated as *P < 0.05, **P < 0.01, and ***P < 0.001.

Back to article page