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目的研究苯代谢产物1,2,4-苯三醇对K562细胞的增殖和分化的影响。方法应用台盼蓝染色排除法计数活细胞数,应用碘化丙啶(PI)染色结合流式细胞术分析细胞周期分布情况,应用PI/FITC-annexin V双染结合流式细胞术分析细胞凋亡情况,联苯胺染色法计数分析K562细胞红系分化情况。结果1,2,4-苯三醇(0.025～0.4 mmol/L)处理24 h对K562细胞产生浓度依赖性的增殖抑制。0.2 mmol/L的1,2,4-苯三醇作用24 h后,K562细胞G0/G1期细胞比例显著下降,S期和G2/M期细胞比例显著升高,细胞出现明显的细胞凋亡。用0.005 mmol/L浓度的1,2,4-苯三醇预处理24h的K562细胞经氯化高铁血红素诱导24、48和72 h的红细胞分化率分别是21.57%、40.62%和46.60%,均显著低于对照细胞相应时间点的红细胞分化率。结论在本试验条件下,苯代谢物1,2,4-苯三醇在一定的浓度范围内对K562细胞表现出浓度依赖性的增殖抑制作用,这种增殖抑制作用可能是通过细胞周期改变进而引起细胞凋亡来实现的,而在无细胞毒性的低浓度下1,2,4-苯三醇就可明显抑制K562细胞的红系分化,1,2,4-苯三醇的凋亡诱导作用和分化抑制作用可能在苯血液毒性机制方面有重要意义。
Objective To study the effect of benzene metabolite 1,2,4-benzenetriol on the proliferation and differentiation of K562 cells. Methods The number of viable cells was counted by trypan blue exclusion method. The cell cycle distribution was analyzed by propidium iodide (PI) staining combined with flow cytometry. The cell apoptosis was analyzed by PI / FITC-annexin V double staining combined with flow cytometry Mortality, benzidine staining counts erythroid differentiation K562 cells. Results Treatment of K562 cells with 1,2,4-benzenetriol (0.025-0.4 mmol / L) for 24 h resulted in a concentration-dependent inhibition of proliferation. After treated with 0.2 mmol / L 1,2,4-benzenetriol for 24 h, the proportion of cells in G0 / G1 phase of K562 cells decreased significantly, the proportion of cells in S phase and G2 / M phase increased significantly, and the cells showed obvious apoptosis . The differentiation rate of erythrocytes of K562 cells pretreated with 0.005 mmol / L 1,2,4-benzenetriol for 24 h, 48 h and 72 h after induction by hemin were 21.57%, 40.62% and 46.60%, respectively, Were significantly lower than the control cells at the appropriate time points erythroid differentiation rate. Conclusion Under the experimental conditions, the benzene metabolite 1,2,4-benzenetriol showed a concentration-dependent proliferation inhibition effect on K562 cells in a certain concentration range, which may be caused by the cell cycle change Induced cell apoptosis, whereas 1,2,4-benzenetriol at a low cytotoxicity concentration significantly inhibited erythroid differentiation of K562 cells and induced apoptosis of 1,2,4-benzenetriol The role of differentiation and inhibition may have important significance in benzene hematological toxicity mechanism.