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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。
Objective: To investigate the effect of high concentration of ascorbic acid (AA) on the proliferation and apoptosis of human ovarian cancer cell line SKOV-3 and its possible mechanism. Methods: The proliferation of SKOV-3 cells was detected by CCK-8 assay after different concentrations of AA. The apoptosis rate and mitochondrial membrane potential of SKOV-3 cells were detected by FCM after treated with 5 mmol / L AA. After staining with DCFH-DA (2 ’, 7’-dichlorofluorescin diacetate), the cells were observed under a fluorescence microscope The expression of reactive oxygen species (ROS) in cells was detected by Western blotting. The expressions of cytochrome C, cleaved-caspase 3 and Bcl-2 were detected by Western blotting. Results: AA could significantly inhibit the proliferation of SKOV-3 cells and had a dose-response relationship (P <0.05). After 5 mmol / L AA treatment, the apoptosis rate of SKOV-3 cells was higher than that of the blank control group (SKOV-3 cells without any intervention) (P <0.05), while the mitochondrial membrane potential decreased more than that of the blank control group (P <0.05), and the expression level of reactive oxygen species in SKOV-3 cells was higher than that in the blank control group (P <0.05). The expressions of cytochrome C and cleaved-caspase 3 protein in SKOV-3 cells treated with 5mmol / 0.05). The expression of Bcl-2 protein was down-regulated (P <0.05). CONCLUSION: AA can effectively inhibit the proliferation of SKOV-3 cells and induce the apoptosis of ovarian cancer SKOV-3 cells, which may lead to mitochondrial apoptosis through ROS generation.