饮水消毒副产物二氯乙腈对HepG2细胞周期的影响

罗皓; 杜进林; 唐焕文; 翟璐; 程小广; 杨慧; 陈佳佳

doi:10.13421/j.cnki.hjwsxzz.2015.03.004

饮水消毒副产物二氯乙腈对HepG2细胞周期的影响

Effects of Dichloroacetonitrile on HepG2 Cells Cycle

摘要

摘要:
目的研究p53基因和chk1基因在饮水消毒副产物二氯乙腈(dichloroacetonitrile, DCAN)对HepG2细胞周期阻滞过程中的影响。
方法以二甲基亚砜(DMSO)溶解DCAN, 以DMSO处理人肝癌细胞(HepG2细胞)为对照组, 分别以低、高浓度(50、400 μmol/L)DCAN染毒HepG2细胞24 h。采用碘化丙啶(PI)标记的流式细胞技术检测细胞周期分布情况, 实时荧光定量聚合酶链反应(qRT-PCR)检测p53基因和chk1基因mRNA表达的改变。
结果与对照组比较, 400 μmol/L处理组G₁期细胞比例明显减少, G₂期细胞比例明显增加, 差异均有统计学意义(P < 0.05);400 μmol/L DCAN作用于HepG2细胞后下调了p53的表达, 差异有统计学意义(P < 0.05), 而对chk1的表达没有影响。
结论高浓度的DCAN能明显延长HepG2细胞的G₂期, 并可抑制p53的mRNA表达水平, 但对chk1的mRNA表达水平无影响。

Abstract:
Objective To explore the role of p53 and chk1 in the cell cycle arrest induced by dichloroacetonitrile (DCAN) in human hepatocellular carcinoma cell line HepG2 cells.
Methods HepG2 cells were exposed to 50 and 400 μmol/L DCAN in Dimethyl sulfoxide (DMSO) for 24 h, and HepG2 cells treated with DMSO only were served as the control. The distribution of cells in different phases of cell cycle was tagged by propidium iodide (PI) staining and determined by flow cytometry kits; the expressions of p53 and chk1 mRNA was quantified by reverse transcription real-time polymerase chain reaction assay.
Results Compared with the control group, the proportion of G₁-phase cells in the 400 μmol/L DCAN-treated group was decreased, while the proportion of G₂-phase cells was increased (P < 0.05). The expression of p53 mRNA was down-regulated in the 400 μmol/L DCAN-treated group (P < 0.05), but no significant effect was observed on the expression of chk1 mRNA.
Conclusion High concentration of DCAN may arrest HepG2 cells at G₂ phase, inhibit the expression of p53 mRNA, but not affect the expression of chk1 mRNA.

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