Fig. 5. The ATP mimic BzATP significantly increased the therapeutic efficacy of chemotherapy in a colorectal cancer animal model.

A HCT116 cells were treated with TNFα (50 ng/mL) and OXP (25 μM) for 24 h. Then, these cells were cocultured with THP1-iDCs (THP1-siNC and THP1-siP2RX7) for 24 h. B The mRNA levels of the DC maturation marker CD86 and the proinflammatory cytokines IL-6, IL-1β and IL-18 in THP-iDCs that were cocultured with vehicle- or OXP/TNFα-treated HCT116^shNC and HCT116^shPANX1 cells were evaluated by qRT‒PCR (n = 3). ***p < 0.001. One-Way ANOVA test. C Schematic diagram of the chemotherapeutic regimen. D A total of 5 × 10⁵ CT26 cells were subcutaneously inoculated into the left sides of the backs of BALB/c mice for 7 days, and the mice were administered FOLFOX (20 μM levofolinate, 20 μM 5-Fu, 100 μM OXP), in combination or not with the ATP agonist BzATP (20 μM, i.p. injection) or the ATP antagonist A438079 (20 μM, i.p. injection) on the indicated days. Tumor volume was measured every 3 days. **p < 0.01 and ***p < 0.001. Two-Way ANOVA test. E The resected tumors were weighted on Day 26 (mean ± s.e.m., n = 5). **p < 0.01 and ***p < 0.001. One-Way ANOVA test. F The level of cleaved caspase-3 and PANX1 within resected tumors was analyzed by western blotting (n = 3). *p < 0.05 and n.s. = not significant. One-Way ANOVA test. G The representative results of CD3⁺, CD8⁺, GzmB⁺CD8⁺ T cells was shown. H The quantification of intratumoral CD3 expression is shown (mean ± s.e.m., n = 3). *p < 0.05 and **p < 0.01. One-Way ANOVA test. I The quantification of intratumoral CD8 expression is shown (mean ± s.e.m., n = 3). *p < 0.05 and **p < 0.01. One-Way ANOVA test. J The quantification of peri-tumoral GzmB⁺CD8⁺ expression is shown (mean ± s.e.m., n = 3). *p < 0.05 and **p < 0.01. One-Way ANOVA test. K The quantification of intratumoral GzmB⁺CD8⁺ expression is shown (mean ± s.e.m., n = 3). *p < 0.05 and **p < 0.01. One-Way ANOVA test.