厚朴酚对人乳腺癌MCF-7细胞增殖和程序性坏死的影响OACSTPCD

Objective To investigate the effect of magnolol on the cell proliferation and the necroptosis of human breast cancer cells and its possible mechanism.Methods Human breast cancer cell line MCF-7 cells were cultured in vitro.The MCF-7 cells were treated with 0,5,10,20,40,80 μmol/L magnolol for 24,48,72 h,or treated with 40 μmol/L magnolol with or without necroptosis inhibitor Nec-1(40 μmol/L)for 24 h,and then the cell viability was detected by MTT assay.The MCF-7 cells were treated with 0,2,4,8 μmol/L magnolol for 10 d,and then the colony-forming ability of MCF-7 cells was detected by colony-formation experiments.After the MCF-7 cells were treated with 40 µmol/L magnolol for 24 h,the morphological changes of MCF-7 cells were observed under transmission electron microscope.The MCF-7 cells were treated with 0,10,20,40,80 µmol/L magnolol for 24 h,and then the necrop-tosis and ROS level were detected by flow cytometry,and the protein expressions of RIP1,RIP3,p-RIP3,MLKL,and p-MLKL were detected by Western blot.Results Compared with 0 μmol/L magnolol,5,10,20,40,80 μmol/L magnolol significantly inhibited MCF-7 viability in a dose dependent manner(P＜0.01).The viability of MCF-7 cells was higher in magnolol+Nec-1 group than in magnolol group(P＜0.01).Compared with 0 µmol/L magnolol,2,4,8 μmol/L magnolol inhibited the cell colony formation.The formation of vacuoles and the vacuoles break were observed under transmission electron microscope after treatment with 40 µmol/L mag-nolol for 24 h.Compared with 0 µmol/L magnolol,the necroptosis rate,the ROS level and the protein levels of RIP1,RIP3,p-RIP3,MLKL and p-MLKL were significantly upregulated after treatment with 10,20,40,80 μmol/L magnolol(P＜0.01).Conclusion Magnolol can induce the necroptosis of human breast cancer MCF-7 cells,which is related to the activation of RIP3-MLKL signaling pathway.