[关键词]
[摘要]
目的 研究粉防己碱对人乳腺癌MCF-7细胞线粒体能量代谢的影响,探讨其抗肿瘤作用及其机制。方法 采用MTT法检测粉防己碱对MCF-7细胞的细胞毒活性,评价其增殖抑制作用;通过检测线粒体膜电位(mitochondrial membrane potential,MMP)变化、三磷酸腺苷(adenosine triphosphate,ATP)含量、活性氧(reactive oxygen species,ROS)水平和呼吸电子传递链复合酶I~IV活力,评价粉防己碱对MCF-7细胞线粒体功能的影响;运用细胞能量代谢分析仪对活体细胞能量代谢进行实时全过程分析,评价粉防己碱对MCF-7细胞能量代谢的影响。结果 粉防己碱可抑制MCF-7细胞增殖,其半数抑制浓度(50% concentration of inhibition,IC50)值为20.13μmol/L;粉防己碱显著降低MCF-7细胞MMP、ATP含量、ROS水平和呼吸电子传递链复合酶I~IV活力(P<0.05、0.01);粉防己碱可对MCF-7细胞能量代谢过程中基础氧消耗、ATP产能、线粒体储备能和质子漏产生影响。结论 粉防己碱能够通过破坏线粒体功能并减少细胞能量代谢,从而抑制MCF-7细胞增殖。
[Key word]
[Abstract]
Objection To study the effect of tetrandrine on mitochondrial energy metabolism of MCF-7 cells, explore the antitumor effect and mechanism of tetrandrine. Methods Cytotoxicity of tetrandrine on MCF-7 cells was measured to evaluate cell proliferation inhibition by MTT assay. The changes of mitochondrial membrane potential (MMP), adenosine triphosphate (ATP) content, reactive oxygen species (ROS) level and respiratory electron transport chain complex enzyme I-IV activities were measured to evaluate the effect of tetrandrine on mitochondrial function. The real-time metabolic process of MCF-7 cells were analyzed by Seahorse Extracellular Analyzer to evaluate the effect of tetrandrine on energy metabolism. Results Tetrandrine inhibited the proliferation of MCF-7 cells, with the 50% concentration of inhibition (IC50) was 20.13 μmol/L. Tetrandrine significantly reduced MMP, ATP content, ROS level and respiratory electron transport chain complex enzyme I-IV activities (P < 0.05, 0.01). Tetrandrine affected basal oxygen consumption, ATP production, mitochondrial reserve energy and proton leakage in the process of energy metabolism in MCF-7 cells. Conclusion Tetrandrine can inhibit MCF-7 cells proliferation by damaging mitochondrial function and reducing cell energy metabolism.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金资助项目(81803686);国家自然科学基金配套项目(2018PP01);教育部“春晖计划”合作科研项目(HLJ2019035);黑龙江省普通本科高等学校青年创新人才培养计划项目(UNPYSCT-2020225);黑龙江省博士后专项经费资助(LBH-Q20180)