于大永, 高鸿雁, 曹鹤, 卢轩, 史丽颖. Chaetoglobosins E诱导乳腺癌MCF-7细胞凋亡作用机制研究[J]. 华南师范大学学报(自然科学版), 2022, 54(1): 61-69. doi: 10.6054/j.jscnun.2022010
引用本文: 于大永, 高鸿雁, 曹鹤, 卢轩, 史丽颖. Chaetoglobosins E诱导乳腺癌MCF-7细胞凋亡作用机制研究[J]. 华南师范大学学报(自然科学版), 2022, 54(1): 61-69. doi: 10.6054/j.jscnun.2022010
YU Dayong, GAO Hongyan, CAO He, LU Xuan, SHI Liying. The Effect of Chaetoglobosins E on the Proliferation and Apoptosis of MCF-7 Cells and the Related Mechanisms[J]. Journal of South China Normal University (Natural Science Edition), 2022, 54(1): 61-69. doi: 10.6054/j.jscnun.2022010
Citation: YU Dayong, GAO Hongyan, CAO He, LU Xuan, SHI Liying. The Effect of Chaetoglobosins E on the Proliferation and Apoptosis of MCF-7 Cells and the Related Mechanisms[J]. Journal of South China Normal University (Natural Science Edition), 2022, 54(1): 61-69. doi: 10.6054/j.jscnun.2022010

Chaetoglobosins E诱导乳腺癌MCF-7细胞凋亡作用机制研究

The Effect of Chaetoglobosins E on the Proliferation and Apoptosis of MCF-7 Cells and the Related Mechanisms

  • 摘要: 为探究Chaetoglobosins E (ChE)对肿瘤细胞增殖和凋亡的影响,体外培养人乳腺癌MCF-7细胞、人膀胱癌T-24细胞、人黑色素瘤C8161细胞、人白血病U937细胞,用不同浓度的ChE分别作用于4种细胞24 h或48 h,MTT法检测4种肿瘤细胞的增殖情况;为进一步研究其作用机制,Hoechst 33342染色观察MCF-7经ChE处理后细胞形态的变化,流式细胞术检测MCF-7经ChE处理后细胞凋亡、周期、活性氧以及线粒体膜电位的变化情况,Western Blot法检测MCF-7细胞中凋亡相关蛋白的表达情况。结果显示:ChE对MCF-7、T-24、C8161和U937细胞增殖均有抑制作用,且均呈现出时间和剂量依赖性,4种肿瘤细胞中,ChE对MCF-7增殖的抑制效果最强,24、48 h的IC50分别为82.04±7.01、49.87±2.28 μmol/L;Hoechst 33342染色发现,随着ChE浓度的升高,凋亡的MCF-7细胞数逐渐增多,细胞凋亡特征显著,细胞核的体积缩小,细胞核裂解并伴有凋亡小体;通过流式细胞术发现,MCF-7细胞经ChE处理后, 细胞凋亡增加、细胞周期改变、活性氧增加以及线粒体膜电位降低;Western Blot实验发现,Bid、Caspase 3蛋白的表达量降低,Cleaved Caspase 3、Bax蛋白与Bcl-2蛋白表达量的比值增加。综上所述,ChE诱导的MCF-7细胞凋亡与Caspase依赖性线粒体途径有关。

     

    Abstract: To explore the effect of Chaetoglobosins E (ChE) on tumor cell proliferation and apoptosis, human breast cancer MCF-7 cells, human bladder cancer T-24 cells, human melanoma C8161 cells and human leukemia U937 cells were treated with ChE of different concentrations for 24 h or 48 h. The MTT method was used to detect the proliferation of 4 kinds of tumor cells. To further study its mechanism of action, the Hoechst 33342 staining was used to observe the changes in the morphology of MCF-7 cells treated with ChE. Flow cytometry was used to detect the changes of apoptosis, cycle, reactive oxygen species and mitochondrial membrane potential of MCF-7 cells treated with ChE. The Western Blot method was used to detect the expression of apoptosis-related proteins in MCF-7 cells. The results showed that ChE inhibited the proliferation of MCF-7, T-24, C8161 and U937 cells in a time- and dose-dependent manner. Among the four tumor cells, ChE had the strongest inhibitory effect on the proliferation of MCF-7 with IC50 of 82.04±7.01 μmol/L and 49.87±2.28 μmol/L at 24 h and 48 h, respectively. The Hoechst 33342 staining found that the number of apoptotic MCF-7 cells gradually increased, with significant characteristics of cell apoptosis and the volume of the nucleus was reduced, with the nucleus lysed and accompanied by apoptotic bodies with the increase of ChE concentration. The flow cytometry found that, after MCF-7 cells were treated with ChE, the number of apoptosis cells increased, the cell cycle was changed, the reactive oxygen species increased, and the mitochondrial membrane potential decreased. The Western Blot experiment found that the expression of Bid and Caspase 3 protein decreased and cleaved Caspase 3 and the ratio of Bax protein to Bcl-2 protein expression increased. To sum up, ChE-induced apoptosis of MCF-7 cells is related to the Caspase-dependent mitochondrial pathway.

     

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