Curcumol induces cell cycle arrest in colon cancer cells via reactive oxygen species and Akt/ GSK3β/cyclin D1 pathway
Curcumol induces cell cycle arrest in colon cancer cells via reactive oxygen species and Akt/ GSK3β/cyclin D1 pathway
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姜黄酚通过活性氧和 Akt/GSK3 beta/cyclin D1 途径诱导结肠癌细胞的细胞周期停滞
DOI：
10.1016/j.jep.2017.06.037
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发表时间：
2018-01-10
影响因子：
5.4
通讯作者：
Chen, Xu
中科院分区：
文献类型：
--
作者：
Wang, Juan;Li, Xu-mei;Chen, Xu
民族药理学相关性：姜黄（Curcuma kwangsiensis s.g. Lee & c.f. Liang）属于姜科植物，作为传统中药已有近2000年的历史。Curcumol是从药用植物Curcuma kwangsiensis s.g. Lee和c.f. Liang中分离得到的一种愈创烷型倍半萜类半晶化合物，具有抗癌作用。我们前期的研究发现，抑制鼻咽癌细胞生长的贡献最大的是姜黄酚。目的：评价姜黄酚对人结肠癌细胞（CRC） LoVo和SW480细胞周期阻滞的作用，探讨其体内外作用机制。材料与方法：莪术酚溶于无水乙醇。对照组或实验样品中无水乙醇的浓度始终为最终培养基体积的1/500 （v/v）。不同浓度姜黄酚（0、53、106、212和424 μ M）处理LoVo和SW480细胞。然后流式细胞术检测各组细胞周期。Western blot检测大鼠外周血中PI3K、p-Akt、cyclin D1、cyclin E、CDK2、CDK4、GSK3 β的表达水平。real-time RT-PCR检测各组细胞中PI3K、Akt、cyclin D1、CDK4、P27、p21、P16 mRNA的表达。此外，姜黄酚还对裸鼠原位肿瘤植入物的抗肿瘤活性进行了评价。结果：莪术醇诱导细胞周期阻滞于G1/S期。RT-qPCR和Western blot数据显示，姜黄酚可增强GSK3 β、P27、p21和P16的表达，降低PI3K、磷酸化Akt （p-Akt）、cyclin D1、CDK4、cyclin E和CDK2的水平。此外，姜黄酚诱导LoVo细胞产生活性氧（ROS），活性氧清除剂n -乙酰半胱氨酸（NAC）显著逆转姜黄酚诱导的细胞生长抑制。此外，姜黄酚还能抑制人结肠癌细胞在裸鼠体内的异种移植物的生长，并伴有PI3K、Akt、cyclin D1、CDK4、cycln E的降低，GSK3 β的显著升高。结论：姜黄酚通过ROS的产生和Akt/ GSK3 β /cyclin D1通路的失活，使细胞周期阻滞在GO/G1期，提示姜黄酚具有预防结肠癌发生的潜力。
Ethnopharmacological relevance: Curcuma kwangsiensis S. G. Lee & C. F. Liang (Guangxi ezhu, in Chinese) belongs to the Zingiberaceae family, has been used as a traditionally Chinese medicine nearly 2000 year. Curcumol is one of the guaiane-type sesquiterpenoid hemiketal isolated from medicine plant Curcuma kwangsiensis S. G. Lee & C. F. Liang, which has been reported possesses anti-cancer effects. Our previous study found that the most contribution to inhibit nasopharyngeal carcinoma cell growth was curcumol.Aim of the study: To assess the effect of curcumol on cell cycle arrest against human colon cancer cells (CRC) cells (LoVo and SW480) and explore its mechanism in vitro and in vivo.Materials and methods: Curcumol was dissolved in absolute ethyl alcohol. The concentration of absolute ethyl alcohol in the control group or in experimental samples was always 1/500 (v/v) of the final medium volume. LoVo and SW480 cells were treated with different concentrations of curcumol (0, 53, 106, 212 and 424 mu M). And then the cell cycle of each group was examined by flow cytometry. The protein levels of PI3K, p-Akt, cyclin D1, cyclin E, CDK2, CDK4 and GSK3 beta were determined by Western blot. The mRNA expression of PI3K, Akt, cyclin D1, CDK4, P27, p21, and P16 in the treated cells were analyzed by real-time RT-PCR. In addition, the antitumor activity of curcumol was evaluated in nude mice bearing orthotopic tumor implants.Results: Curcumol induced cell cycle arrest in G1/S phase. RT-qPCR and Western blot data showed that curcumol enhanced the expression of GSK3 beta, P27, p21 and P16, and decreased the levels of PI3K, phosphorylated Akt (p-Akt), cyclin D1, CDK4, cyclin E and CDK2. Furthermore, curcumol induced reactive oxygen species (ROS) generation in LoVo cells, and ROS scavenger N-acetylcysteine (NAC) significantly reversed curcumol-induced cell growth inhibition. Besides, curcumol also prevented the growth of human colon cancer cells xenografts in nude mouse, accompanied by the reduction of PI3K, Akt, cyclin D1, CDK4, cycln E and significant increase of GSK3 beta.Conclusions: Curcumol caused cell cycle arrest at the GO/G1 phase by ROS production and Akt/ GSK3 beta/cyclin D1 pathways inactivation, indicating the potential of curcumol in the prevention of colon cancer carcinogenesis.