Fig. 6

Combination treatment with GSK461364 and Enz in PCa cells. A, B The viability of C4-2 and 22Rv1 PCa cells was assessed via the CCK-8 assay following 48 h of treatment with GSK461364, Enz, or their combination. C Cell death was evaluated via propidium iodide (PI) staining and fluorescence microscopy after 24 h of treatment. Scale bar, 100 μm. D Colony formation assays were performed on treated C4-2 and 22Rv1 cells after 14 days, with colony counts quantified via ImageJ software. E PI staining and bright-field imaging of PCa organoids were conducted to assess cell death following treatment with GSK461364, Enz, or their combination. Scale bar, 50 μm. In nude mice, the combination of Enz and GSK461364 more effectively inhibited tumor growth. At the end of the experiment, the mice were euthanized, and F Representative tumor images are shown. G Tumor growth curves were recorded every three days (mean ± SD, n = 5/group), and tumor tissues were weighed and summarized. H Representative H&E and IHC staining of the indicated proteins in tumor tissues from each group. Scale bar, 100 μm. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test (G). G Data are presented as mean ± SD. Representative data of triplicate experiments are shown. *p < 0.05, **p < 0.01, ***p < 0.001, ns, not significant