摘要
目的:探究贝母素乙(Peiminine)对乳腺癌细胞MCF-7细胞凋亡的影响及其可能作用机制。方法:采用不同浓度贝母素乙或联合PI3K抑制剂LY294002干预MCF-7细胞,MTT法检测细胞增殖能力;Hoechst 33258染色和Annexin V-FITC/PI流式细胞术检测细胞凋亡情况;JC-1染色法检测细胞线粒体膜电位变化;Western blotting检测细胞中PI3K(p110α)、Akt、p-Akt(ser473)、Bad、Bax、Bcl-2、cleaved-Caspase-3以及线粒体和胞浆中细胞色素C(Cyt C)等蛋白表达水平。结果:贝母素乙可呈时间-浓度依赖性抑制MCF-7细胞增殖,诱导细胞出现凋亡形态改变,促进细胞凋亡,并降低线粒体膜电位,上调细胞中Bad、Bax、cleaved-Caspase-3及胞浆中Cyt C蛋白表达水平,下调PI3K(p110α)、p-Akt、Bcl-2和线粒体中Cyt C蛋白表达水平,而Akt蛋白表达水平无显著变化。然而,联合LY294002干预可增强贝母素乙对MCF-7细胞凋亡的促进作用。结论:贝母素乙可诱导乳腺癌MCF-7细胞凋亡,可能是通过阻断PI3K/Akt信号通路介导的线粒体凋亡途径实现的。
Objective:To investigate the effect of peiminine on apoptosis of breast cancer MCF-7 cells and its possible mechanism.Methods:MCF-7 cells were treated with different concentrations of peiminine or combined with PI3K inhibitor LY294002.MTT assay was used to detect the cell proliferation.Hoechst 33258 staining and Annexin V-FITC/PI flow cytometry assay were used to detect the cell apoptosis.JC-1 staining assay was used to detect mitochondrial membrane potential change.Western blotting was used to detect the protein expression of levels of PI3K(p110α),Akt,p-Akt(ser473),Bad,Bax,Bcl-2,cleaved-Caspase-3 in cells and Cyt C in mitochondrial and cytoplasm.Results:Peiminine inhibited MCF-7 cells proliferation in a time and concentration dependent manner.Meanwhile,peiminine induced morphological changes of apoptosis,promoted apoptosis,reduced mitochondrial membrane potential,and up-regulated the protein expression levels of Bad,Bax,cleaved-Caspase-3,and Cyt C in cytoplasm,down-regulated the protein expression levels of PI3K,p-Akt,Bcl-2,and Cyt C in mitochondrial.There was no significant effect on the expression level of Akt protein.However,peiminine combined with LY294002 intervention can enhance the promotion of MCF-7 cells apoptosis.Conclusion:Peiminine can promote the apoptosis of breast cancer MCF-7 cells,which may be achieved by blocking the mitochondrial apoptosis pathway mediated by PI3K/Akt signaling pathway.
作者
易念
王莉
王松
包铮
胡聂
李莉红
蒋忠军
徐舸
YI Nian;WANG Li;WANG Song;BAO Zheng;HU Nie;LI Lihong;JIANG Zhongjun;XU Ge(Department of Thyroid and Breast Surgery,the Affiliated Nanhua Hospital,Hengyang Medical School,University of South China,Hunan Hengyang 421002,China)
出处
《现代肿瘤医学》
CAS
北大核心
2023年第12期2180-2185,共6页
Journal of Modern Oncology
基金
湖南省自然科学基金(编号:2020JJ4552)
湖南省科技厅临床医疗技术创新引导计划(编号:2017SK50217)
湖南省衡阳市科技局指导性项目(编号:114)。
