Figures & data
Figure 1. HPP inhibited the proliferation of bladder cancer cells in the co-culture system. A CCK-8 assay. B-C EdU staining assay (magnification, ×400). The fluorescence of EdU (red) and Hoechst (blue) represent proliferating cells and cell nuclei, respectively. D CCK-8 assay. *p < 0.05; **p < 0.01; ***p < 0.001 compared with the control group.
Figure 2. HPP induced bladder cancer cell autophagy in the co-culture system. A-B Protein expression of p62, Beclin1, and LC3II/I was determined by Western blotting analysis. C-F MB49 cells were pre-treated with 5 mM 3-MA for 4 h or 50 μM CQ for 3 h, and then co-cultured with RAW264.7 cells in fresh medium containing 100 μg/mL HPP. Protein expression of LC3II/I was determined by Western blotting analysis. *p < 0.05; **p < 0.01; ***p < 0.001 compared with the control group. ##p < 0.01 compared with the HPP group.
Figure 3. HPP induced bladder cancer cell autophagy by regulating macrophages in the co-culture system, resulting in the inhibition of cancer cell proliferation. A the formation of autophagosomes in MB49 cells was observed by MDC staining (magnification, ×200) and TEM (scale bar, 2 μm). The red arrows indicate autophagosomes. B-C CCK-8 assay. D-G Protein expression of LC3II/I was determined by Western blotting analysis. *p < 0.05; **p < 0.01; ***p < 0.001 compared with the control group. #p < 0.05; ##p < 0.01 compared with the HPP group.
Figure 4. PI3K/Akt/mTOR signaling pathway was involved in HPP-induced autophagy in the co-culture system. A-B Protein expression of PI3K, Akt, p-Akt, mTOR, and p-mTOR was determined by Western blotting analysis. *p < 0.05; **p < 0.01; ***p < 0.001 compared with the control group.
Data availability statement
The data supporting the findings of this study are available from the corresponding author upon reasonable request.