【摘要】 目的:探讨高浓度抗坏血酸(ascorbic acid,AA)对人卵巢癌SKOV-3细胞增殖和凋亡的影响及其可能的作用机制。方法:不同浓度的AA作用后,CCK-8法检测SKOV-3细胞的增殖情况。5 mmol/L AA处理SKOV-3细胞后,应用FCM法检测细胞的凋亡率和线粒体膜电位的变化情况;DCFH-DA(2’,7’-dichlorofluorescin diacetate)染色后,在荧光显微镜下观察细胞内活性氧的表达水平;蛋白质印迹法检测细胞内cytochrome C、cleaved-caspase 3和Bcl-2蛋白的表达情况。结果:AA能显著抑制SKOV-3细胞的增殖,且有一定的剂量-效应关系(P<0.05)。5 mmol/L AA处理后,SKOV-3细胞的凋亡率高于空白对照组(未进行任何干预的SKOV-3细胞)(P<0.05),而线粒体膜电位的下降率高于空白对照组(P<0.05),活性氧的表达水平高于空白对照组(P<0.05);5mmol/L AA处理组SKOV-3细胞中cytochrome C和cleaved-caspase 3蛋白的表达水平上调(P值均<0.05),Bcl-2蛋白的表达水平下调(P<0.05)。结论:AA能有效抑制人卵巢癌SKOV-3细胞的增殖,诱导细胞凋亡,并可能通过产生活性氧使细胞发生线粒体途径的凋亡。更多还原

【Abstract】 Objective:To investigate the effects of ascorbic acid(AA) on the proliferation and apoptosis of human ovarian cancer SKOV-3 cells,and to explore the possible mechanism.Methods:The proliferation of SKOV-3 cells after treatment with different concentrations of AA was detected by CCK-8 assay.After treatment with 5 mmol/L AA,the changes of apoptotic rate and mitochondrial membrane potential of SKOV-3 cells were measured by FCM,and the expression of reactive oxygen species stained by 2’,7’-dichlorofluorescin diacetate was examined under a fluorescence microscope,while the expressions of cytochrome C,cleaved-caspase 3 and Bcl-2 proteins were detected by Western blotting.Results:AA could inhibit the proliferation of SKOV-3 cells in a dose-dependent manner(P <0.05).The apoptotic rate of SKOV-3 cells after 5 mmol/L AA treatment was higher than that of the blank control(SKOV-3 cells without any treatment)(P < 0.05).The descent rate of mitochondrial membrane potential was higher than that of the blank control(P < 0.05),the expression level of reactive oxygen species was higher than that of the blank control(P <0.05).The expression levels of cytochrome C and cleaved-caspase 3 in SKOV-3 cells in AA treatment group were up-regulated(both P < 0.05),and the expression level of Bcl-2 protein was down-regulated(P < 0.05).Conclusion:AA can inhibit the proliferation and induce apoptosis of human ovarian cancer SKOV-3 cells.The apoptosis may be induced by reactive oxygen species through mitochondrial pathway.更多还原