Abstarct: Using cold plasma reduces cell proliferation, binding and migration and induces apoptosis in neoplastic cells, while show no damaging effect on healthy cells. Therefore, it can be a good therapeutic treatment compared to other treatments for cancer cells. In this study, helium plasma jet was used to generate create plasma at room temperature as a Spot radiation for the treatment of breast cancer cells (MCF_7) with a frequency of 20 kHz, voltage 5 kv, flow 2.5 L/min in time 120 seconds with Three different doses (duty cycle 3,4,5). Using the cytotoxicity test (MTT), the percent survival of the treated cells was examined. Also, Migration Assay and Real Time PCR tests were performed and the results were evaluated by ELISA Reader. According to the results, in MTT experiment at duty cycle = 5 μsec , the best effect of plasma and the highest apoptosis was observed, also showed the highest increase in apoptosis in 72 hours after treatment. But in the cell migration test, the results of the treatment indicated an increase in cell migration compared to the control group. Real Time PCR results indicate that Bax gene (Gene induced apoptosis) exxpression was higher in Duty cycle = 5 μsec, which means that it has the most induction of apoptosis.