Colorectal cancer has a high incidence rate with significant mortality and morbidity. The 15-lipoxygenase- 1 (15-LOX-1) is a crucial enzyme as well as a tumor suppressor, especially in colon cancer. Bee venom, by the induction of apoptosis, is a promising new anti-cancer agent. Thus, the effects of bee venom on the expression of 15-LOX-1 m-RNA and its activities in the HT-29 cell line have been investigated in the current study. Human HT-29 colon cancer cells were treated with increasing concentrations of bee venom (1.87-30 μg/mL), and cisplatin (5 μg/mL) as the positive control for 24- and 48-h. Then, cell viability and apoptosis were measured using the MMT assay and flow cytometry, respectively. Enzyme activity and expression were assayed using the kinetic method and real-time PCR, respectively. The results showed that the main content of bee venom was with a retention time of 60 min. The IC50 values of bee venom at 24- and 48-h were 6.01 μg/mL and 4.44 μg/mL, respectively. Expression of 15-LOX-1 in cancer cells treated with bee venom increased (p < 0.0005), as well as the activity of the enzyme in the presence of bee venom (p < 0.01). The current study revealed the apoptotic and cytotoxic effects of bee venom against the human colon cancer HT-29 cell line that was not seen in fibroblast cells. Findings suggest that bee venom may have therapeutic effects against the HT- 29 colon cancer cell through the induction of the15-LOX-1 pathway. However, further studies are needed in this regard.