Abstract: Nanosecond pulsed electric field (nsPEF) has been demonstrated to induce cancerous cell apoptosis, inhibit tumor growth and elicit antitumor immunity. Caused by the apoptosis mechanism of nanosecond pulse stimulation (NPS) therapy involving too many targets and the limitation of conventional detecting specific biomarker technology, the apoptosis mechanism of NPS has not been reached consensus and there is no efficient method for researching it. In order to gain further insight into the apoptosis mechanism, the paper has studied the murine melanoma B16 cells stimulated by nsPEF by using antibody array. A hypothesis for the pathway of NPS with three steps for the apoptosis mechanism was introduced. The research method possess the unique advantages in this kind of researches with huge number of targets, the obtained results could be helpful for designing future NPS therapies and aid in targeting the specific molecules with the optimal pulse parameters to obtain best therapeutic effect.