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Key Points: Physical activity is associated with reduced mortality rates for survivors of colorectal cancer. Acute high intensity interval exercise (HIIE) reduced colon cancer cell number in vitro and promoted increases in inflammatory cytokines immediately following exercise. This acute suppression of colon cancer cell number was transient and not observed at 120 minutes post-acute HIIE. The acute effects of exercise may constitute an important mechanism by which exercise can influence colorectal cancer outcomes., Abstract: Physical activity is associated with significant reductions in colorectal cancer mortality. However, the mechanisms by which exercise mediates this anti-oncogenic effect are not clear. In the present study, colorectal cancer survivors completed acute (n = 10) or chronic (n = 10) exercise regimes. An acute high intensity interval exercise session (HIIE; 4 × 4 min at 85-95% peak heart rate) was completed with serum samples collected at baseline, as well as 0 and 120 min post-exercise. For the 'chronic' intervention, resting serum was sampled before and after 4 weeks (12 sessions) of HIIE. The effect of serum on colon cancer cell growth was evaluated by incubating cells (CaCo-2 and LoVo) for up to 72 h and assessing cell number. Serum obtained immediately following HIIE, but not 120 min post-HIIE, significantly reduced colon cancer cell number. Significant increases in serum interleukin-6 (P = 0.023), interleukin-8 (P = 0.036) and tumour necrosis factor-α (P = 0.003) were found immediately following acute HIIE. At rest, short-term HIIE training did not promote any changes in cellular growth or cytokine concentrations. The acute effects of HIIE and the cytokine flux may be important mediators of reducing colon cancer cell progression. Repetitive exposure to these acute effects may contribute to the relationship between exercise and improved colorectal cancer survival., (© 2019 The Authors. The Journal of Physiology © 2019 The Physiological Society.)