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目的:探讨2-脱氧-D-葡萄糖(2-deoxy-D-glucose,2-DG)对肺腺癌A549细胞增殖的影响及其相关机制。方法:不同浓度的2-DG与抗氧化剂N-乙酰-L-半胱氨酸(N-acetyl-L-cysteine,NAC)、超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)单独及联合处理A549细胞,CCK-8(cell counting kit-8)法检测细胞生存率;DCFH-DA(2’,7’-dichlorodihydro uorescein diacetate)染色检测细胞内活性氧(reactive oxygen species,ROS)水平的变化;比色法检测A549细胞内氧化型谷胱甘肽占总谷胱甘肽含量的百分比;实时荧光定量PCR检测细胞内SOD、CAT和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)mRNA的转录情况。结果:2-DG可以显著抑制A549细胞的生长,同时伴随着细胞内ROS产生增多和氧化型谷胱甘肽所占的百分比升高。2-DG干预后,细胞内CAT和GSH-Px mRNA表达水平上调,而SOD mRNA的表达水平未出现明显变化。A549细胞经NAC和CAT预处理后,2-DG的抗肿瘤作用明显减弱,而特异性氧自由基清除剂SOD对2-DG的抗肿瘤作用没有明显影响。结论:2-DG能够抑制A549细胞增殖,其机制可能与诱导ROS的生成,尤其是H2O2的生成有关。
Objective: To investigate the effect of 2-deoxy-D-glucose (2-DG) on the proliferation of lung adenocarcinoma A549 cells and its related mechanism. Methods: The effects of different concentrations of 2-DG and antioxidants N-acetyl-L-cysteine (NAC), superoxide dismutase (SOD) and catalase A549 cells were treated with catalase (CAT) alone and in combination with cell counting kit-8 (CCK-8). Cell viability was measured by DCFH-DA staining with 2’7’-dichlorodihydro uorescein diacetate (ROS) were detected by colorimetric assay. The percentage of oxidized glutathione to total glutathione in A549 cells was detected by colorimetric assay. The activities of SOD, CAT and glutathione peroxidase (glutathione peroxidase, GSH-Px) mRNA transcription. Results: 2-DG can significantly inhibit the growth of A549 cells, accompanied by increased intracellular ROS production and the percentage of oxidized glutathione increased. After intervention with 2-DG, the expression of CAT and GSH-Px mRNA in the cells increased, while the expression of SOD mRNA did not change significantly. The antitumor effect of 2-DG was significantly attenuated in A549 cells pretreated with NAC and CAT, while the specific oxygen free radical scavenger SOD had no obvious effect on the antitumor effect of 2-DG. Conclusion: 2-DG can inhibit the proliferation of A549 cells. The mechanism may be related to the induction of ROS production, especially the generation of H2O2.