Figure 6. Compound 10 as a potent anticancer agent for HER2-positive gastric cancer cells.

(A) Apoptosis induced by compound 10 in parental NCI-N87 was assessed by treatment with the compound in a dose-dependent manner (24 hr treatment at indicated concentrations, n=3, mean ± S.D., ANOVA, ns = non-significant, ***p<0.001 vs. CON). (B) Changes in the pro-apoptotic markers were evaluated by treating compound 10 in dose- and time-dependent manner. (C) Anti-proliferative effect of compound 10 (10 μM) was evaluated against NCI-N87 cell line (10 days of treatment at indicated concentrations, n=3, mean ± S.D., ANOVA, ***p<0.001 vs. CON). (D) Tumor growth inhibitory effect of compound 10 was evaluated using NCI-N87 xenograft mouse model (n=5 per group; intravenous (IV) injection at 4 mg/kg every 3 days, indicated by red arrows). Tumor volumes were evaluated at the indicated time points by measuring the length and width of the tumor with calipers using the equation (length x width²)/2. Data was indicated as mean ± S.E.M. (E) Photograph of the tumors collected from the vehicle- and compound 10-treated mice. (F) Final volume changes were assessed for the tumors excised from each experimental group (n=5, mean ± S.E.M., Student’s t-test, ***p<0.001 vs. CON) (G) IHC analysis was conducted against HER2 and Ki67 in the tumors (scale bar = 100 μm). Score quantification was performed using Image J software (10 independent fields per sample were evaluated, mean ± S.D., Student’s t test, ***p<0.001 vs. CON).