Figures & data
Figure 1. The effects of combined ENZ and ATO on the proliferation of CRPC cells and inducing enhanced apoptosis. A-B. The antiproliferative effect of ENZ and ATO in combination was evaluated by colony formation assay. C-E. FACS analysis of C4-2B cells with different treatments and quantification of cell cycle distribution. F. DNA fragmentation was analyzed using ELISA detection. The data are presented as mean ± SD of three independent measurements performed in triplicate. **** p < 0.0001, *** p < 0.001, ** p < 0.01.
Figure 2. Abnormal upregulation of DDR-related pathways was involved in the progression of PCa. A. The DEGs of DDR-related pathways and the targets of ATO were derived from the Drugbank database, and visualized in the heatmap with GSE32269 data. B. Significant upregulation of the TOPBP1-ATR-CHEK1-CDC25C signaling in metastatic CRPC compared to primary PCa. **** p < 0.0001, ** p < 0.01.
Figure 3. Involvement of DDR-related signalings in the effect of ENZ combination with ATO on CRPC cells. A. The DEGs of the DDR-related pathways and the targets of ATO were derived from the Drugbank database, and visualized by the heatmap with the RNA-seq library data. B. The frequency of the genes was visualized with the R package "wordcloud2". The larger the sample size of the word, the higher the frequency. C. The fold change of the expression levels of genes was visualized with the volcano plot. D-E. The expression levels of DNA damage and apoptosis-related genes of C4-2B cells following different treatments. ****p < 0.0001, ns p > = 0.05.
Figure 4. The combination of ENZ and ATO repressed angiogenic activity of C4-2B cells. A-B. The expression levels of angiogenesis-related genes were calculated by WB analysis. ****p < 0.0001, ***p < 0.001, **p < 0.01.
Figure 5. The combination of ENZ and ATO inhibited the growth of C4-2B xenografts. A. The in vivo protocol of the experimental design and treatment groups. B-D. Significant differences were noted in the volume and wet weight of the C4-2B xenografts with different groups. The data are presented as mean ± SD, ****p < 0.0001.
Figure 6. The effects of ENZ and ATO co-treatment on the angiogenesis, apoptosis and DNA repair processes in vivo. A-D. These effects were monitored on C4-2B xenografts following different treatments and evaluated with immunohistochemical analysis. A. Apoptosis was evaluated with TUNEL staining. B. VEGFR2 expression visualized with IHC staining. C. Ki-67 expression visualized with IHC staining. D. HR repair was assessed for BRAC1. **** p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns p > = 0.05.
Data availability statement
The data that support the findings of this study are available from the corresponding author, Luo Y, upon reasonable request.