1,4-苯醌致K562细胞线粒体功能障碍与凋亡作用

目的：研究1,4-苯醌对人慢性髓系白血病细胞（K562细胞）线粒体功能及凋亡的影响。方法：分别用终浓度为0、10、20 μmol/L的1,4-苯醌处理K562细胞24 h，用CCK-8法检测细胞活力，通过流式细胞仪检测活性氧（ROS）生成量；用线粒体膜电位、三磷酸腺苷（ATP） 生成量来评价线粒体功能；用PI-Annexin V双染法检测细胞的凋亡，采用分光光度法检测caspase-3酶的活性。结果：与对照组比较，1,4-苯醌10和20 μmol/L染毒组K562细胞相对增殖率均降低（P均<0.05）；随着1,4-苯醌浓度的增加，ROS生成量逐渐上升、线粒体膜电位和ATP生成量均逐渐降低、细胞的凋亡率逐渐增高，其中1,4-苯醌20 μmol/L染毒组与对照组间的差异均有统计学意义（P < 0.05或P < 0.01）；caspase-3活性逐渐升高，1,4-苯醌10和20 μmol/L染毒组与对照组相比差异均有统计学意义（P均<0.01）。结论：1,4-苯醌可以诱导K562细胞ROS升高，抑制K562细胞增殖，造成线粒体功能障碍，诱导细胞凋亡升高，提示线粒体障碍在1,4-苯醌诱导K562细胞凋亡的过程中发挥了重要作用。

Induction of mitochondrial dysfunction and apoptosis by 1,4-benzoquinone in K562 cells

OBJECTIVE:To determine effects of 1,4-BQ on mitochondrial dysfunction and apoptosis in human chronic myeloid leukemia cells (K562 cells).METHODS:K562 cells were treated with 0,10,and 20μmol/L 1,4-BQ for 24 h. Cell viability was detected by CCK-8 assay,production of reactive oxygen species (ROS) was detected by flow cytometry;function of mitochondria was evaluated by measuring mitochondrial membrane potentialand adenosine triphosphate (ATP);and caspase-3 enzyme activity was detected using spectrophotometry. RESULTS:Compared with the control,the relative growth rate of K562 cells in the 1,4-BQ 10 and 20μmol/L treatment groups decreased with the increased concentration of 1,4-BQ (P<0.05). Production of ROS and cell apoptosis rates were elevated whilemitochondrial membrane potential and the amounts of ATPwere reduced. Between the 1,4-BQ 20μmol/L exposure groups and the control group,the difference was statistically significant (P<0.05 orP<0.01). The activity of caspase-3 was increased,and the difference was statistically significant (P<0.01) between the control and both 1,4-BQ treatment groups. CONCLUSION:1,4-BQ induced increase of ROS in K562 cells,inhibited their proliferation and resulted in mitochondrial dysfunction and expression of apoptosis. This indicates that mitochondrial dysfunction was involved with 1,4-BQ-induction of apoptosis in K562 cells.