[ effect of Honey bee venom on induction of differentiation of k562 cell line to erythroid linage]

ABSTRACT

Complementary medicine uses bee venom [BV] to treat several diseases, including arthritis and skin diseases. BV contains mellitin, phospholipase A2, apamin and several other bioactive substances. According to the venom compounds, the purpose of this study was to examine the effects of BV on differentiation of K562cell line. In this experimental study, K562cells were treated with different doses of BV in different durations. BV toxicity was evaluated by MTT assay. Benzidine staining was used to investigate the effects of BV on K562 cell differentiation toward the erythroid line. In order to determine the type of cell death, annexin-V gene expression was analyzed by flow cytometry. Colony assay was used to measure BV ability in inhibiting colony formation. Morphological changes in the cells undergone treatment with BV were evaluated by wright-giemsa staining. MTT assay showed that bee venom with concentrations of 5.5-6 microg/ml and 3.5-4.5microg/ml result in 505 cell death in 24h and 48h, respectively. Morphological examination and benzidine staining showed that lower doses in longer period induce differentiation in these cells. Flow cytometry data showed significantly increased in annexin-V gene expression in cells which were treated with bee venom for 24 h. Colony assay demonstrated that the concentration of 1 microg/ml of BV results in 50% reduction in colony formation