Bu Ming, Ren Wen-Kang, Wang Lu, Sun Chun-Xue, Luo Ran, Guo Xiao-Shan, Lin Yu, Ge Peng-Ling, Liu Ji-Cheng
Postdoctoral Research Station, Qiqihar Institute of Medical and Pharmaceutical Sciences Qiqihar 161006, China Postdoctoral Research Station, Heilongjiang University of Chinese Medicine Harbin 150040, China School of Pharmacy, Qiqihar Medical University Qiqihar 161006, China.
School of Pharmacy, Qiqihar Medical University Qiqihar 161006, China.
Zhongguo Zhong Yao Za Zhi. 2024 Aug;49(15):4139-4147. doi: 10.19540/j.cnki.cjcmm.20240402.403.
This study aims to explore the effect and mechanism of a mitochondrion-targeted derivative of ergosterol peroxide(Mito-EP) on breast cancer. The methyl thiazolyl tetrazolium(MTT) assay was employed to examine the proliferation of MDA-MB-231 cells treated with different concentrations(0, 0.075, 0.15, 0.3, 0.6, 1.2, and 2.4 μmol·L(-1)) of Mito-EP. Cells were grouped for treatment with water(blank control), low, medium, and high concentrations(0.15, 0.3, and 0.6 μmol·L(-1)) of Mito-EP, and ergosterol peroxide(EP)(0.6 μmol·L(-1)). After the cells were treated for 48 h, flow cytometry was employed to examine the apoptosis rate, reactive oxygen species(ROS) level, mitochondrial membrane potential, and cell cycle distribution, and the apoptosis, ROS, and mitochondrial membrane potential were observed by laser confocal microscopy. A mouse model bearing subcutaneous xenograft tumor was established by injecting 4T1 cell suspension and used to study the inhibitory effect of Mito-EP on breast cancer. Western blot was employed to determine the protein levels of B-cell lymphoma 2(Bcl-2), Bcl-2-associated X protein(Bax), cytochrome C(Cyt C), cleaved caspase-7, and cleaved caspase-9 in cells and the tumor tissue. The results showed that Mito-EP reduced the proliferation rate of MDA-MB-231 cells in a concentration-dependent manner. Compared with the blank control group, EP(0.6 μmol·L(-1)) caused slight changes in the apoptosis rate, ROS level, and mitochondrial membrane potential. However, Mito-EP increased the apoptosis rate, elevated the ROS level, decreased mitochondrial membrane potential, up-regulated the protein levels of Bax, Cyt C, cleaved caspase-7, and cleaved caspase-9, and down-regulated the protein level of Bcl-2(all P<0.05). Moreover, Mito-EP reduced the tumor volume and weight. In summary, Mito-EP may promote apoptosis in breast cancer cells by activating the mitochondrial apoptosis pathway.
本研究旨在探讨麦角甾醇过氧化物的线粒体靶向衍生物（线粒体 - 麦角甾醇过氧化物，Mito - EP）对乳腺癌的作用及机制。采用甲基噻唑基四氮唑蓝（MTT）法检测不同浓度（0、0.075、0.15、0.3、0.6、1.2和2.4 μmol·L⁻¹）的Mito - EP处理后MDA - MB - 231细胞的增殖情况。将细胞分为用水（空白对照）、低、中、高浓度（0.15、0.3和0.6 μmol·L⁻¹）的Mito - EP以及麦角甾醇过氧化物（EP，0.6 μmol·L⁻¹）处理的组。细胞处理48小时后，采用流式细胞术检测凋亡率、活性氧（ROS）水平、线粒体膜电位和细胞周期分布，并通过激光共聚焦显微镜观察凋亡、ROS和线粒体膜电位。通过注射4T1细胞悬液建立荷皮下异种移植瘤小鼠模型，用于研究Mito - EP对乳腺癌的抑制作用。采用蛋白质免疫印迹法检测细胞和肿瘤组织中B细胞淋巴瘤2（Bcl - 2）、Bcl - 2相关X蛋白（Bax）、细胞色素C（Cyt C）、裂解的半胱天冬酶 - 7和裂解的半胱天冬酶 - 9的蛋白水平。结果表明，Mito - EP以浓度依赖性方式降低MDA - MB - 231细胞的增殖率。与空白对照组相比，EP（0.6 μmol·L⁻¹）对凋亡率、ROS水平和线粒体膜电位有轻微影响。然而，Mito - EP增加了凋亡率，提高了ROS水平，降低了线粒体膜电位，上调了Bax、Cyt C、裂解的半胱天冬酶 - 7和裂解的半胱天冬酶 - 9的蛋白水平，并下调了Bcl - 2的蛋白水平（均P < 0.05）。此外，Mito - EP减小了肿瘤体积和重量。综上所述，Mito - EP可能通过激活线粒体凋亡途径促进乳腺癌细胞凋亡。
