目的 研究石蒜碱介导人乳腺癌MDA-MB-231细胞自噬及凋亡的作用机制。方法 采用CCK-8法检测石蒜碱对MDA-MB-231细胞增殖的影响；倒置显微镜、荧光显微镜和激光共聚焦扫描显微镜（laser scanning confocal microscopy，LSCM）观察细胞形态；细胞集落实验检测细胞克隆形成能力；划痕实验检测细胞迁移能力；流式细胞仪检测细胞凋亡率、细胞内活性氧（reactive oxygen species，ROS）水平、线粒体膜电位；LSCM检测线粒体膜通透性转换孔（mitochondrial permeability transition pore，MPTP）开放情况；Western blotting检测线粒体自噬相关蛋白［线粒体内膜转位酶（translocase of inner membrane，TIM）、线粒体外膜转位酶（translocase of outer membrane，TOM）、E3泛素连接酶（E3 ubiquitin protein ligase，PARKIN）、PTEN诱导的激酶1（PTEN induced putative kinase 1，PINK1）、微管相关蛋白轻链3（microtubule-associated protein light chain 3，LC3-B）、p62］和凋亡相关蛋白［半胱氨酸天冬氨酸蛋白酶-3（cystein-asparate protease-3，Caspase-3）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、细胞色素C（cytochrome-C，Cyt-C）］的表达。结果 石蒜碱对MDA-MB-231细胞具有生长抑制作用，其半数抑制浓度（half inhibitory concentration，IC₅₀）值为6.21 μmol/L；石蒜碱能够改变细胞形态，部分细胞出现凋亡迹象；石蒜碱能够显著抑制细胞的增殖和迁移能力（P＜0.01）；与对照组比较，随着石蒜碱给药浓度的增加，细胞凋亡率随之升高（P＜0.01），ROS水平升高（P＜0.01），线粒体膜电位下降（P＜0.01），MPTP开放（P＜0.01）；石蒜碱给药48 h后，细胞内TIM、TOM、p62和Bcl-2蛋白表达量下调（P＜0.01），PARKIN、PINK1、LC3-B、Caspase-3、Bax和Cyt-C蛋白表达量上调（P＜0.01），且呈浓度相关性。结论 石蒜碱对MDA-MB-231细胞具有生长抑制作用，并可通过调控线粒体氧化损伤介导MDA-MB-231细胞的自噬及凋亡。

Objective To investigate the mechanism of lycorine on autophagy and apoptosis in human breast cancer MDA-MB-231 cell. Methods The effect of lycorine on MDA-MB-231 cell proliferation was detected by CCK-8; Cell morphology was observed by inversion microscopy, fluorescence microscopy and laser scanning confocal microscopy (LSCM); The cell clone formation ability was detected by cell colony assay; The cell migration ability was examined by scratch assay; The apoptosis rate, intracellular reactive oxygen species (ROS) level and mitochondrial membrane potential were determined by flow cytometry; The opening of mitochondrial membrane permeability transition pore (MPTP) was detected by LSCM; The expressions of mitochondrial autophagy-related proteins [translocase of inner membrane (TIM), translocase of outer membrane (TOM), E3 ubiquitin protein ligase (PARKIN), PTEN induced putative kinase 1 (PINK1), microtubule-associated protein light chain 3 (LC3-B), p62], and apoptosis-related proteins [cystein-asparate protease-3 (Caspase-3), Bcl-2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), cytochrome-C (Cyt-C)] were detected by Western blotting. Results Lycorine had a proliferative inhibitory effect on MDA-MB-231 cells, with a half inhibitory concentration (IC₅₀) value of 6.21 μmol/L; Lycorine could change cell morphology, and some cells showed signs of apoptosis; Lycorine could significantly inhibit the proliferation and migration capacity of MDA-MB-231 cells (P < 0.01); Compared with control group, with the increase of lycorine administration concentration, cell apoptosis rate was increased (P < 0.01), ROS level was increased (P < 0.01), mitochondrial membrane potential was decreased (P < 0.01), and MPTP was open (P < 0.01). After 48 h treatment of lycorine, the expressions of TIM, TOM, p62 and Bcl-2 protein were down-regulated (P < 0.01), and the expressions of PARKIN, PINK1, LC3-B, Caspase-3, Bax and Cyt-C were up-regulated (P < 0.01) in a concentration-dependent manner. Conclusion Lycorine has growth inhibition effect on MDA-MB-231cells, and induces autophagy and apoptosis of MDA-MB-231 cells through the regulation of mitochondrial oxidative damage.

R285.5

中国博士后面上项目（2021MD703828）；黑龙江省自然科学基金优秀青年项目（YQ2022H002）；哈尔滨商业大学产业化项目支持计划项目（XL0086）；2023年度哈尔滨商业大学镜湖学者支持计划项目（JHQNRC01）；哈尔滨市科技局项目（2023ZCZJCG038）；哈尔滨商业大学成果转化项目（2023-KYYWF-1034）