Objective To explore the regulation of survivin and RASSF2 genes and apoptosis of breast cancer cells by neferine. Methods The breast cancer cell line Zr-75-30 purchased from ATCC cell bank was divided into cancer cell group (CC group), low concentration group (LC group), medium concentration group (MC group) and high concentration group (HC group). In CC group, Zr-75-30 cells were cultured normally without any other treatment. In LC group, Zr-75-30 cells were mixed with 5 mg/kg neferine. In MC group, Zr-75-30 cells were mixed with 10 mg/kg neferine. In HC group, Zr-75-30 cells were mixed with 20 mg/kg neferine. Flow cytometry, Hoechst 33258 fluorescence staining, Western blot, immunohistochemistry and qRTPCR were used to detect the apoptosis, survivin protein expression, RASSF2 expression, survivin and RASSF2 mRNA expression of Zr-75-30 cells. Results In CC group, the fluorescence intensity of Zr-75-30 cells was weak and the number of apoptotic cells was less. The probability of nuclear pyknosis in LC group, MC group and HC group was significantly higher than that in CC group (all P＜0.05). Compared with LC group and MC group, the number of apoptotic cells in HC group was the most (all P＜0.05). The expression of survivin protein in Zr-75-30 cells of HC group was the lowest. The expression of survivin protein in Zr-75-30 cells of CC group was higher than that of LC group, MC group and HC group (all P＜0.05). The expression of survivin protein in LC group was higher than that in MC group and HC group (all P＜0.05). RASSF2 was positive in Zr-75-30 cells and showed purple granules. The number of rassf2 positive cells in HC group was higher than that in CC group. The number of rassf2 positive cells in HC group was significantly higher than that in LC and MC groups (all P＜0.05). The expression of survivin mRNA was the lowest in HC group and the highest in CC group. Compared with LC group and MC group, the expression of survivin mRNA in HC group was significantly lower (all P＜0.05), and the expression of RASSF2 mRNA in HC group was opposite to that in MC group. Conclusion Neferine (Nef) has a promoting effect on the apoptosis of breast cancer (zr-75-30) cells, and it has a positive correlation with the concentration. The promoting effect is the best at a concentration of 20 mg/kg, and its principle of action It may be related to Nef inhibiting survivin expression and up-regulating RASSF2 expression in cells.