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目的研究维生素C对PM2.5致人肺癌A549细胞周期阻滞的影响。方法用设定浓度的PM2.5(50、100和200μg/ml)单独或联合20μg/ml维生素C分别处理A549细胞24 h。用MTT比色法检测细胞活力,荧光探针检测细胞内活性氧,试剂盒检测MDA和SOD变化,用流式细胞仪检测细胞的周期时相,用Western blot检测细胞周期调控蛋白GADD45α的表达量。结果与空白对照组相比,PM2.5(50、100和200μg/ml)可降低细胞活力,提高ROS和MDA含量,降低SOD酶活,并在100和200μg/ml浓度组提高GADD45α蛋白表达量,使A549细胞阻滞在G2期。而20μg/ml维生素C可显著抑制上述损伤指标的升高,提高SOD酶活性,降低G2期细胞阻滞。结论本实验初步证明PM2.5可引起A549细胞氧化损伤并导致细胞发生G2期阻滞,维生素C抗氧化剂可有效抑制这一过程的发生,对细胞起到保护作用。
Objective To investigate the effect of vitamin C on cell cycle arrest of human lung cancer A549 cells induced by PM2.5. Methods A549 cells were treated with PM2.5 (50, 100 and 200 μg / ml), alone or in combination with 20 μg / ml vitamin C for 24 h. Cell viability was detected by MTT colorimetric assay, reactive oxygen species (ROS) were detected by fluorescent probe, the changes of MDA and SOD were detected by kit, the cell cycle phase was detected by flow cytometry, the expression of GADD45α . Results Compared with the blank control group, PM2.5 (50, 100 and 200 μg / ml) decreased cell viability, increased ROS and MDA content, decreased SOD activity, and increased the expression of GADD45α protein at 100 and 200 μg / ml , A549 cells arrested in the G2 phase. While 20μg / ml vitamin C can significantly inhibit the above indicators of injury, improve SOD activity, reduce G2 cell arrest. Conclusion This experiment proves that PM2.5 can cause oxidative damage of A549 cells and lead to cell arrest in G2 phase. Vitamin C antioxidant can effectively inhibit this process and play a protective role on cells.