线粒体ROS改变参与石蒜碱诱导的HepG2细胞凋亡-《南京师范大学学报》（自然科学版）

文章信息/Info

Title:: Mitochondrial ROS Contributes to Lycorine-inducedApoptosis in HepG2 Cells

Author(s):: Kang Jingjing; Liu Xiaoning; Department of Biochemistry,School of Medicine,Huanghe Science and Technology College,Zhengzhou 450063,China

摘要:: 探究石蒜碱诱导HepG2细胞凋亡的分子机理. CCK-8实验检测细胞增殖情况; 流式细胞术检测加或不加SS31情况下,石蒜碱对HepG2细胞凋亡比例的影响; 用核酸染料Hoechst 33342评估凋亡细胞细胞核的形态变化; 用Western blot分析技术检测PARP片段的表达情况. 成功构建靶向线粒体的Mito-Grx1-roGFP2转基因稳定株肝癌细胞系HepG2. SS31作为线粒体ROS的定向清除剂,能够改善石蒜碱引起的HepG2细胞形态学损伤及细胞核内PARP切割,抑制石蒜碱诱导的细胞凋亡,

Abstract:: To further explore the underlying apoptotic molecular mechanism of lycorine on HepG2 cells. The growth of HepG2 cells were measured with TransDetect Cell Counting Kit(CCK-8)assay. Apoptosis percentages of cells exposed to lycorine with or without SS31 was determined by flow cytometry. Apoptotic nuclear morphology was assessed using Hoechst 33342. Western blot analysis was used to examine the expression of fragmented PARP. It was successfully established that an human hepatoma cell line HepG2 with mito-Grx1-roGFP2 probe stably expressed which could be used as a real-time reporter of dynamic redox changes. As an antioxidant which could remove the ROS of mitochondria,the treatment of SS31 improved the morphology of HepG2 cells injured by lycorine and inhibited lycorine-induced cell apoptosis. It was also found that SS31 significantly inhibited the fragment of PARP induced by lycorine. Furthermore,SS31 attenuated lycorine-induced rise of 405/488 fluorescence ratio which could reflect the level of ROS in mitochondria. The findings suggest that mitochondrial ROS contribute to lycorine-induced apoptosis in HepG2 cells and lycorine is expected to be a potential drug for the treatment of liver cancer,it worth profund study.