Polychlorinated biphenyls (PCBs) are industrial compounds, which were used prevalently from the 1930s until the mid-1970s. Aroclor 1242 (A1242), a commercial mixture of PCBs, were produced by Monsanto Chemical Company of St. Louis, Missouri and contains 42% chlorine. Previous studies suggested that PCBs inhibitory effect on reproductive function, developmental abnormality, and impaired reproductive ability and they have also effects disrupting the endocrine system such as decreasing testosterone synthesis and steroidogenic enzyme activities in Leydig cells. The present study was aimed at determining the direct effects of A1242 on Leydig cell viability, oxidative damage and steroidogenic enzyme profile in vitro and attenuating effects of two antioxidants, vitamin E (VitE) and genistein (Gen), against this damage. TM3 Leydig cells were exposed to A1242 (10-8 and 10-6 mol/L), in addition to these concentrations, to VitE (50µM) and Gen (10µM) as antioxidant for 24 h. Following the exposure time, the Leydig cells were evaluated for measurement of cell viability assay, lipid peroxidation assay, reactive oxygen species (ROS) and steroidogenic enzymes. The results showed that cell viability was reduced after A1242 exposure, while lipid peroxidation and ROS increased. Furthermore, steroidogenesis was interrupted in a concentration-dependent manner. Following A1242, administrations of VitE or Gen as an antioxidant reduce hazard effects of A1242 on Leydig cells. In conclusion, our results imply that exposure to A1242 may impair Leydig cell function and cause toxicity in Leydig cells. VitE and Gen treatment also exhibited therapeutic effects against this toxicity.