In our previous study, it was reported that 2[[3-(2,3-dichlorophenoxy)propyl]amino]ethanol (2,3-DCPE) induces apoptosis and cell cycle arrest. The current study aimed to investigate the molecular mechanism involved in 2,3-DCPE-induced S phase arrest. The results demonstrated that 2,3-DCPE upregulated phosphorylated (p-)H2A histone family member X, a biomarker of DNA damage, in the DLD-1 colon cancer cell line. Western blotting revealed that 2,3-DCPE increased the checkpoint kinase (Chk)1 (Ser317 and Ser345) level and decreased the expression of M-phase inducer phosphatase 1 (Cdc25A) in a time-dependent manner. Subsequently, the results demonstrated that the ataxia-telangiectasia mutated (ATM) and ataxia-telangiectasia and Rad3-related (ATR) inhibitors wortmannin and caffeine had no effect on the cell cycle; however, the inhibitors partially abrogated 2,3-DCPE-induced S phase arrest. Flow cytometry assays revealed that caffeine (2 mM) reduced the proportion of S phase cells from 83 to 39.6% and that wortmannin (500 nM) reduced the proportion of S phase cells from 83 to 48.2%. Furthermore, wortmannin and caffeine inhibited the 2,3-DCPE-mediated phosphorylation of Chk1 and the degradation of Cdc25A. However, these ATM/ATR inhibitors had limited effect on 2,3-DCPE-induced apoptosis. Taken together, the data of the current study indicated that 2,3-DCPE caused DNA damage in colon cancer cells and that 2,3-DCPE-induced S phase arrest was associated with the activation of the ATM/ATR-Chk1-Cdc25A pathway.

中文翻译：

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小分子2,3-DCPE通过激活DLD-1结肠癌细胞中的ATM / ATR-Chk1-Cdc25A信号传导途径诱导S期阻滞。

在我们先前的研究中，据报道2 [[3-（2,3-二氯苯氧基）丙基]氨基]乙醇（2,3-DCPE）诱导细胞凋亡和细胞周期停滞。当前的研究旨在调查参与2，3-DCPE诱导的S期阻滞的分子机制。结果表明，在DLD-1结肠癌细胞系中，2,3-DCPE上调了磷酸化（p-）H2A组蛋白家族成员X（DNA损伤的生物标记）。Western印迹显示，2，3-DCPE以时间依赖的方式增加了检查点激酶（Chk）1（Ser317和Ser345）的水平，并降低了M期诱导磷酸酶1（Cdc25A）的表达。随后，结果表明共济失调-毛细血管扩张突变（ATM）和共济失调-毛细血管扩张和Rad3相关（ATR）抑制剂渥曼青霉素和咖啡因对细胞周期没有影响。然而，抑制剂部分废除了2，3-DCPE诱导的S期阻滞。流式细胞仪检测显示，咖啡因（2 mM）将S期细胞的比例从83％降至39.6％，而渥曼青霉素（500 nM）将S期细胞的比例从83至48.2％。此外，渥曼青霉素和咖啡因抑制2，3-DCPE介导的Chk1磷酸化和Cdc25A的降解。但是，这些ATM / ATR抑制剂对2,3-DCPE诱导的细胞凋亡的作用有限。综上所述，本研究的数据表明，2,3-DCPE引起结肠癌细胞DNA损伤，并且2,3-DCPE诱导的S期阻滞与ATM / ATR-Chk1-Cdc25A途径的激活有关。 。6％，而渥曼青霉素（500 nM）将S期细胞的比例从83％降至48.2％。此外，渥曼青霉素和咖啡因抑制2，3-DCPE介导的Chk1磷酸化和Cdc25A的降解。但是，这些ATM / ATR抑制剂对2,3-DCPE诱导的细胞凋亡的作用有限。综上所述，本研究的数据表明，2,3-DCPE引起结肠癌细胞DNA损伤，并且2,3-DCPE诱导的S期阻滞与ATM / ATR-Chk1-Cdc25A途径的激活有关。 。6％，而渥曼青霉素（500 nM）将S期细胞的比例从83％降至48.2％。此外，渥曼青霉素和咖啡因抑制2，3-DCPE介导的Chk1磷酸化和Cdc25A的降解。但是，这些ATM / ATR抑制剂对2,3-DCPE诱导的细胞凋亡的作用有限。综上所述，本研究的数据表明，2,3-DCPE引起结肠癌细胞DNA损伤，并且2,3-DCPE诱导的S期阻滞与ATM / ATR-Chk1-Cdc25A途径的激活有关。 。