Fig. 5
From: CRO-67 has anti-cancer activity in pancreatic tumor cells and stromal cancer-associated fibroblasts
CRO-67 activated apoptosis and reduced proliferation in two PDAC cell lines and patient-derived CAFs. PDAC cell lines (MiaPaCa-2 and PANC-1) and primary patient-derived CAFs isolated from N = 5 PDAC patients were treated with vehicle control or CRO-67 (1.5 μM) and analyzed 24 h post-treatment. (a–c) Phase-contrast confluency measurements were taken every hour for 48 h following CRO-67 treatment of PDAC cell lines and CAFs. MiaPaCa-2 (N = 3), PANC-1 (N = 3) PDAC cells and CAFs (N = 5 PDAC patients) demonstrated a decrease in cell proliferation, as measured by the IncuCyte S3 Live Cell Analysis system. (d–f) Apoptosis was measured by flow cytometry for Annexin V/DAPI. MiaPaCa-2 (N = 3), PANC-1 (N = 3) PDAC cells and CAFs (N = 5) demonstrated an increase in apoptosis 24 h post-treatment with CRO-67 (1.5 μM). (g–i) Flow cytometry was used to quantify the percentages of cells in the G1, S, and G2/M phases of the cell cycle at 4 and 24 h post-treatment with CRO-67 (1.5 μM). CRO-67 treatment in MiaPaCa-2 (N = 3), PANC-1 (N = 3) and CAF (N = 3) cells resulted in the accumulation of cells in the G2/M phase at 4 and 24 h post-treatment, suggesting mitotic arrest. Bars represent mean ± SD (N = 3). Asterisks indicate significance as assessed by one-way ANOVA, and Bonferroni’s multiple comparisons tests; performed on endpoint data at 48 h timepoint for cell proliferation (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001).
