Novel microtubule-targeted agent 6-chloro-4-(methoxyphenyl) coumarin induces G2-M arrest and apoptosis in HeLa cells
Novel microtubule-targeted agent 6-chloro-4-(methoxyphenyl) coumarin induces G2-M arrest and apoptosis in HeLa cells
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DOI：
10.1038/aps.2011.176
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发表时间：
2012-03-01
影响因子：
8.2
通讯作者：
Li, Jia
中科院分区：
文献类型：
--
作者：
Ma, Yi-ming;Zhou, Yu-bo;Li, Jia
目的：鉴定一种具有最高抗癌活性的新型香豆素类似物，并进一步探讨其抗癌机制。方法：采用MTT法测定肿瘤细胞活力。使用流式细胞术和Western blotting分析评估细胞周期进展。体内用免疫细胞化学和体外用微管蛋白解聚实验观察微管解聚。膜联蛋白V/碘化丙啶（PI）双染色和亚g(1)分析显示细胞凋亡。结果：在36种香豆素类似物中，6-氯-4-（甲氧基苯基）香豆素对HCT116细胞的抗癌活性最好，IC50值约为200 nmol/L。该化合物对结肠癌、乳腺癌、肝癌、宫颈癌、白血病、表皮样癌等9种肿瘤细胞系具有广谱的抗肿瘤活性，IC50值为75 nmol/L-1.57 μ mol/L，对WI-38人肺成纤维细胞的IC50值为12.128 μ mol/L，细胞杀伤活性较低。化合物（0.04 ~ 10 μ mol/L）诱导HeLa细胞G(2)-M相阻滞呈剂量依赖性，去除化合物后可逆。该化合物（10-300 μ mol/L）在体外诱导纯化的猪微管蛋白解聚。最后，化合物（0.04 ~ 2.5 μ mol/L）诱导HeLa细胞凋亡呈剂量和时间依赖性。结论：6-氯-4-（甲氧基苯基）香豆素是一种新型微管靶向药物，可诱导HeLa细胞G(2)- m阻滞和凋亡。
Aim: To identify a novel coumarin analogue with the highest anticancer activity and to further investigate its anticancer mechanisms.Methods: The viability of cancer cells was investigated using the MTT assay. The cell cycle progression was evaluated using both flow cytometric and Western blotting analysis. Microtubule depolymerization was observed with immunocytochemistry in vivo and a tubulin depolymerization assay in vitro. Apoptosis was demonstrated using Annexin V/Propidium Iodide (PI) double-staining and sub-G(1) analysis.Results: Among 36 analogues of coumarin, 6-chloro-4-(methoxyphenyl) coumarin showed the best anticancer activity (IC50 value about 200 nmol/L) in HCT116 cells. The compound had a broad spectrum of anticancer activity against 9 cancer cell lines derived from colon cancer, breast cancer, liver cancer, cervical cancer, leukemia, epidermoid cancer with IC50 value of 75 nmol/L-1.57 mu mol/L but with low cytotocitity against WI-38 human lung fibroblasts (IC50 value of 12.128 mu mol/L). The compound (0.04-10 mu mol/L) induced G(2)-M phase arrest in HeLa cells in a dose-dependent manner, which was reversible after the compound was removed. The compound (10-300 mu mol/L) induced the depolymerization of purified porcine tubulin in vitro. Finally, the compound (0.04-2.5 mu mol/L) induced apoptosis of HeLa cells in dose-and time-dependent manners.Conclusion: 6-Chloro-4-(methoxyphenyl) coumarin is a novel microtubule-targeting agent that induces G(2)-M arrest and apoptosis in HeLa cells.