Induction of G1 and G2/M cell cycle arrests by the dietary compound 3,3'-diindolylmethane in HT-29 human colon cancer cells.
Induction of G1 and G2/M cell cycle arrests by the dietary compound 3,3'-diindolylmethane in HT-29 human colon cancer cells.
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DOI：
10.1186/1471-230x-9-39
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发表时间：
2009-05-29
影响因子：
2.4
通讯作者：
Park JH
中科院分区：
文献类型：
--
作者：
Choi HJ;Lim DY;Park JH
3，3 '-二吲哚基甲烷（DIM）是食用西兰花和其他十字花科蔬菜后在胃中产生的吲哚衍生物，已被证明在体内和体外模型中发挥抗癌作用。DIM（0 - 30 μmol/L）对HT-29人结肠癌细胞的生长有抑制作用，且呈浓度依赖性。在这项研究中，我们评估了DIM对HT-29细胞周期进程的影响。用不同浓度的DIM（0 ~ 30 μmol/L）培养HT-29细胞，碘化丙啶染色，流式细胞术分析。对细胞周期蛋白依赖性激酶（CDK）和细胞分裂周期（CDC）2进行了[3 H]胸苷掺入试验、Western印迹分析、免疫沉淀和体外激酶试验。DIM处理12 h后，G1期和G2/M期细胞比例呈剂量依赖性增加，S期细胞比例呈剂量依赖性减少。DIM还以剂量依赖性方式减少DNA合成。DIM显著降低CDK 2活性和磷酸化视网膜母细胞瘤蛋白（Rb）和E2 F-1的水平，并增加低磷酸化Rb的水平。DIM降低了细胞周期蛋白A、D1和CDK 4的蛋白水平。DIM还增加了CDK抑制剂p21 CIP 1/WAF 1和p27 KIPI的蛋白水平。此外，DIM降低了CDC 2的活性以及CDC 25 C磷酸酶和细胞周期蛋白B1的水平。在这里，我们已经证明，DIM诱导HT-29细胞G1和G2/M期细胞周期阻滞，这种作用可能是通过降低CDK活性介导的。
3,3'-Diindolylmethane (DIM), an indole derivative produced in the stomach after the consumption of broccoli and other cruciferous vegetables, has been demonstrated to exert anti-cancer effects in both in vivo and in vitro models. We have previously determined that DIM (0 – 30 μmol/L) inhibited the growth of HT-29 human colon cancer cells in a concentration-dependent fashion. In this study, we evaluated the effects of DIM on cell cycle progression in HT-29 cells. HT-29 cells were cultured with various concentrations of DIM (0 – 30 μmol/L) and the DNA was stained with propidium iodide, followed by flow cytometric analysis. [3H]Thymidine incorporation assays, Western blot analyses, immunoprecipitation and in vitro kinase assays for cyclin-dependent kinase (CDK) and cell division cycle (CDC)2 were conducted. The percentages of cells in the G1 and G2/M phases were dose-dependently increased and the percentages of cells in S phase were reduced within 12 h in DIM-treated cells. DIM also reduced DNA synthesis in a dose-dependent fashion. DIM markedly reduced CDK2 activity and the levels of phosphorylated retinoblastoma proteins (Rb) and E2F-1, and also increased the levels of hypophosphorylated Rb. DIM reduced the protein levels of cyclin A, D1, and CDK4. DIM also increased the protein levels of CDK inhibitors, p21CIP1/WAF1 and p27KIPI. In addition, DIM reduced the activity of CDC2 and the levels of CDC25C phosphatase and cyclin B1. Here, we have demonstrated that DIM induces G1 and G2/M phase cell cycle arrest in HT-29 cells, and this effect may be mediated by reduced CDK activity.