Chemical induction of the bystander effect in normal human lymphoblastoid cells

Many studies investigating the bystander effect have used ionizing radiation to evaluate this phenomenon, whereas very few have determined whether genotoxic chemicals are also capable of inducing this effect. Here, we show that two such chemicals, mitomycin C, a bifunctional alkylating agent and phleomycin, a glycopeptide antibiotic of the bleomycin family, cause normal human B lymphoblastoid cells to produce media soluble factors that induce a bystander effect in unexposed cells. Ionizing radiation was used in parallel experiments to verify the existence of the bystander effect in these cells. Micronuclei in Cytochalasin B-blocked binucleated cells were used as the endpoint. Conditioned media obtained from cells exposed to mitomycin C induced a 1.5-3 fold increase, while conditioned media from phleomycin induced a 1.5-4 fold increase, and conditioned media from irradiated cells induced a 2-8 fold increase in micronuclei. We conclude that the bystander effect is not restricted to ionizing radiation, suggesting it may be a part of a general cellular stress response.