Bladder cancer is the most common type of tumor in the urinary system and has a high incidence and mortality rate. Protozoan parasites have recently gained popularity as a research topic for cancer therapy. The present study aimed to evaluate the effect of
T. gondii
tachyzoite culture supernatants on bladder cancer cells. Bladder cancer 5637 cells were infected with two different genotypes of
T. gondii
tachyzoite culture supernatants, RH and ToxoDB#9. The effects of
T. gondii
tachyzoite culture supernatant on cell proliferation were detected using a Cell Counting Kit-8 assay. The effects on cell migration and invasion were evaluated using wound healing and Transwell assays. The effect on the apoptosis rate of the cells was assessed using a TUNEL assay, while the protein levels of Bax and Bcl-2 were analyzed by western blotting. The results indicated that the culture supernatant of
T. gondii
tachyzoites altered the morphology of bladder cancer 5637 cells, inhibited cell proliferation, reduced cell migration and invasion and promoted apoptosis. The ToxoDB#9-type experimental group showed higher efficacy than the RH-type experimental group in inhibiting bladder cancer cell proliferation, reducing cell migration and invasion and promoting apoptosis. These results indicated that the culture supernatant of
T. gondii
tachyzoites could be a promising therapeutic agent for the treatment of bladder cancer.