姜黄素通过活性氧和 Akt/GSK3β/细胞周期蛋白 D1 通路诱导结肠癌细胞周期停滞。
Curcumol induces cell cycle arrest in colon cancer cells via reactive oxygen species and Akt/ GSK3β/cyclin D1 pathway.
机构信息
Xiangya Hospital Central South University, Chang sha 410008, China; College of Pharmacy, Guilin Medical University, Guilin 541004, China.
College of Pharmacy, Guilin Medical University, Guilin 541004, China.
出版信息
J Ethnopharmacol. 2018 Jan 10;210:1-9. doi: 10.1016/j.jep.2017.06.037. Epub 2017 Jul 4.
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 424μM). 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 GSK3β 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 GSK3β, 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 GSK3β.
CONCLUSIONS
Curcumol caused cell cycle arrest at the G0/G1 phase by ROS production and Akt/ GSK3β/cyclin D1 pathways inactivation, indicating the potential of curcumol in the prevention of colon cancer carcinogenesis.
民族药理学相关性
广西莪术（Guangxi ezhu，中文）属于姜科植物，作为一种传统中药已有近 2000 年的历史。莪术醇是从莪术属植物中分离出的一种愈创木烷型倍半萜半缩醛，具有抗癌作用。我们之前的研究发现，莪术醇对抑制鼻咽癌细胞生长的贡献最大。
研究目的
评估莪术醇对人结肠癌（LoVo 和 SW480）细胞周期阻滞的影响，并探讨其在体内外的作用机制。
材料与方法
莪术醇溶于无水乙醇。对照组或实验组中无水乙醇的浓度始终为最终培养基体积的 1/500（v/v）。用不同浓度的莪术醇（0、53、106、212 和 424μM）处理 LoVo 和 SW480 细胞。然后通过流式细胞术检测各组细胞周期。用 Western blot 检测 PI3K、p-Akt、cyclin D1、cyclin E、CDK2、CDK4 和 GSK3β 的蛋白水平。通过实时 RT-PCR 分析处理细胞中 PI3K、Akt、cyclin D1、CDK4、P27、p21 和 P16 的 mRNA 表达。此外，在荷瘤原位移植裸鼠中评估莪术醇的抗肿瘤活性。
结果
莪术醇诱导细胞周期停滞在 G1/S 期。RT-qPCR 和 Western blot 数据显示，莪术醇增强了 GSK3β、P27、p21 和 P16 的表达，降低了 PI3K、磷酸化 Akt（p-Akt）、cyclin D1、CDK4、cyclin E 和 CDK2 的水平。此外，莪术醇诱导 LoVo 细胞产生活性氧（ROS），ROS 清除剂 N-乙酰半胱氨酸（NAC）显著逆转了莪术醇诱导的细胞生长抑制。此外，莪术醇还能防止人结肠癌细胞异种移植物在裸鼠中的生长，同时降低 PI3K、Akt、cyclin D1、CDK4、cyclin E 的表达，显著增加 GSK3β 的表达。
结论
莪术醇通过产生 ROS 和抑制 Akt/GSK3β/cyclin D1 通路失活引起细胞周期停滞在 G0/G1 期，表明莪术醇在预防结肠癌发生方面具有潜力。
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