This study aims to investigate the effect of betulinic acid (BA) on pyroptosis in colorectal cancer SW620 cells and its action mechanism based on the regulation of cysteinyl aspartate-specific proteinase-3 (Caspase-3)/gasdermin E (GSDME) signaling pathway. The effect of BA on the proliferation of SW620 cells was detected by the MTT assay. The effect of BA on the colony formation of SW620 cells was examined by colony formation assay. The influence of BA on the morphology of SW620 cells was observed under a microscope. The influence of BA on the pyroptosis morphology of SW620 cells was appraised with a transmission electron microscope. The effect of BA on the pyroptosis rate of SW620 cells was measured by flow cytometry. The damage of BA to the membrane of SW620 cells was assessed by the lactate dehydrogenase (LDH) kit. The ELISA was used to evaluate the action of BA on the release of inflammatory factors including interleukin-1β (IL-1β) and interleukin-18 by SW620 cells. Western blot was employed to determine the action of BA on the levels of proteins of Caspase-3/GSDME signaling pathway. The immunofluorescence staining was utilized to probe the impact of BA on the GSDME N-terminal (GSDME-N) with pore-forming activity in SW620 cells. Compared with the control, BA at 2.5, 5.0 and 10.0 μmol/L inhibited the proliferation and colony formation of SW620 cells (P<0.05, P<0.01). BA gave rise to the morphological characteristics of pyroptosis in SW620 cells, and promoted the proportion of Annexin V/propidium iodide-double positive cells (P<0.01). Meanwhile, BA significantly enhanced the release of LDH, IL-1β and IL-18 by SW620 cells (P<0.05, P<0.01), suggesting that BA could induce pyroptosis in SW620 cells. Compared with the control, BA also attenuated the GSDME protein level and augmented the protein levels of Cleaved Caspase-3 and GSDME-N in SW620 cells (P < 0.05, P < 0.01). Collectively, BA may inhibit CRC by inducing pyroptosis in SW620 cells through activation of Caspase-3/GSDME signaling pathway.