Whole bee venom (BV) is used to treat inflammatory diseases in Korean traditional medicine. Various studies have demonstrated anti-inflammatory and anticancer effects of BV. The toxicity of individual components of BV has been widely studied, although few studies have reported on the toxicity of BV. We sought to evaluate the cytotoxicity of BV in normal human lymphocytes and HL-60 cells. When cells were treated with BV at concentrations of 1 or 5 microg/ml, BV induced cell death in a time-dependent manner until 24 h, but these cytotoxic effects ended thereafter. When cells were treated with BV at a concentration of 10 microg/ml, however, viability decreased until 72 h, which may have been due to the half-life of BV. Whole BV also inhibited proliferation in these cells. BV induced DNA fragmentation and micronuclei in HL-60 cells and DNA fragmentation in human lymphocytes. Phosphate and tensin homolog (PTEN) up-regulation in HL-60 cells may induce S-phase cell cycle arrest. Forkhead transcription factor (FKHR and FKHRL1) up-regulation in human lymphocytes by whole BV treatment may be involved in the repair of damaged DNA and reduce genotoxicity. Based on these results, whole BV may exert cytotoxicity in these two cells in a different fashion.