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目的研究苯并(a)芘(BaP)对人胚肺成纤维细胞(HELF)的细胞周期分布及丝裂原活化蛋白激酶(MAPK)信号分子(ERK1/2、JNK1/2和 p38)磷酸化水平的影响,并探讨 MAPK 磷酸化水平改变与细胞周期效应之间的关系。方法用0.1、0.5、2.5和12.5 μmol/L 的 BaP 处理 HELF 细胞24 h,用蛋白印迹方法检测 ERK1/2、JNK1/2和 p38蛋白激酶的磷酸化水平和蛋白总量的改变,利用流式细胞技术检测2.5 μmol/L BaP 处理24 h 后对 HELF 细胞周期的影响。结果不同剂量 BaP 可明显提高 ERK1/2、JNK1/2和 p38蛋白激酶磷酸化水平;2.5 μmol/L BaP 处理24 h,细胞周期分布与二甲基亚砜对照组相比,G_0与 G_1期细胞数下降了11.9%(F=41.38,P<0.01),S 期细胞数增加了17.2%(F=68.13,P<0.01);三种 MAPK 化学抑制剂(AG126、SP600125及 SB203580)可明显抑制BaP 处理引起的细胞周期的改变。结论 ERK1/2、JNK1/2和 p38均参与了 BaP 所致细胞周期改变过程,并发挥正性调节作用。
Objective To investigate the cell cycle distribution of human embryonic lung fibroblasts (HELF) and the phosphorylation of mitogen - activated protein kinase (MAPK) signaling molecules (ERK1 / 2, JNK1 / 2 and p38) Level, and to explore the relationship between the changes of MAPK phosphorylation and cell cycle effects. Methods HELF cells were treated with BaP at concentrations of 0.1, 0.5, 2.5 and 12.5 μmol / L for 24 h. The phosphorylation of ERK1 / 2, JNK1 / 2 and p38 protein kinases and the total protein levels were detected by Western blotting. Effect of 2.5 μmol / L BaP on HELF cell cycle after 24 h treatment. Results BaP at different doses significantly increased phosphorylation levels of ERK1 / 2, JNK1 / 2 and p38 protein kinase. After treatment with 2.5 μmol / L BaP for 24 h, the cell cycle distribution was similar to that of dimethyl sulfoxide control group (F = 41.38, P <0.01), and the number of S phase cells increased by 17.2% (F = 68.13, P <0.01). Three MAPK chemical inhibitors (AG126, SP600125 and SB203580) Treatment caused changes in the cell cycle. Conclusions Both ERK1 / 2, JNK1 / 2 and p38 are involved in the cell cycle alteration induced by BaP and play a positive regulatory role.