Lutein inhibits proliferation, invasion and migration of hypoxic breast cancer cells via downregulation of HES1
Lutein inhibits proliferation, invasion and migration of hypoxic breast cancer cells via downregulation of HES1
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DOI：
10.3892/ijo.2018.4332
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发表时间：
2018-06-01
影响因子：
5.2
通讯作者：
Zhang, Shanfeng
中科院分区：
文献类型：
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作者：
Li, Yuan;Zhang, Yuekun;Zhang, Shanfeng
包括乳腺癌在内的实体瘤常可观察到瘤内低氧微环境。叶黄素是一种植物来源的化合物，非维生素A类胡萝卜素，已被证明具有多种保护作用，包括抗炎，抗氧化应激和抗肿瘤作用。本研究的主要目的是阐明叶黄素在低氧条件下产生活性氧(ROS)、毛发和分裂增强子1(HES1)的激活以及乳腺癌细胞的增殖、侵袭和迁移中的作用。将人乳腺癌细胞株MDA-MB-157和MCF-7暴露于低氧条件和不同浓度的叶黄素中。四甲基偶氮唑蓝([3-(4，5-dimethylthiazol-2-yl)-2，5-diphenyltetrazolium)比色法检测细胞增殖，Annexin V-异硫氰酸荧光素/碘化丙啶染色分析细胞凋亡率。逆转录-定量聚合酶链式反应和免疫印迹分析检测缺氧诱导因子-1(HIF-1)、Notch信号分子、HES1和上皮-间充质转化(EMT)相关因子的表达。伤口愈合和Transwell侵袭试验检测乳腺癌细胞的侵袭和迁移。用2，7-二氯二氢荧光素-二乙酸酯和流式细胞仪检测细胞内ROS水平。结果表明，在低氧条件下，叶黄素呈剂量依赖性抑制细胞增殖，细胞凋亡率随着叶黄素处理时间的延长而逐渐升高。叶黄素可抑制缺氧诱导的HIF-1活化、Noch信号转导和HES1表达，并抑制缺氧诱导的EMT相关因子的表达。叶黄素显著抑制低氧条件下乳腺癌细胞的侵袭和迁移。叶黄素也能减少缺氧诱导的ROS的产生。此外，ROS清除剂N-乙酰半胱氨酸还抑制低氧条件下乳腺癌细胞缺氧诱导因子1和HES1的表达，但过氧化氢(H_2O_2)水平升高。综上所述，本研究的结果表明，叶黄素可能是一种新的乳腺癌化学预防候选药物。此外，HES1可能在介导叶黄素参与抑制缺氧驱动的ROS诱导的乳腺癌进展中起关键作用。
An intratumoral hypoxic microenvironment is frequently observed in solid tumors, including breast cancer. Lutein, a plant-derived compound and non-vitamin A carotenoid, has been demonstrated to possess multiple protective properties including anti-inflammation, anti-oxidative stress and antitumor effects. The main objective of the present research was to elucidate the involvement of lutein in the production of reactive oxygen species (ROS) under hypoxia, the activation of hairy and enhancer of split 1 (HES1), and the proliferation, invasion and migration of breast cancer cells. The human breast cancer cell lines MDA-MB-157 and MCF-7 were exposed to hypoxic conditions and various concentrations of lutein. An MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay was performed to examine cell proliferation, and Annexin V-fluorescein isothiocyanate/propidium iodide staining was performed to analyze the apoptosis ratio. The levels of hypoxia inducible factor-1 (HIF-1), NOTCH signaling molecules, HES1 and epithelial-mesenchymal transition (EMT)-associated factors were examined by reverse transcription-quantitative polymerase chain reaction and western blot analysis. Wound healing and Transwell invasion assays were used to detect the invasion and migration of breast cancer cells. Intracellular ROS levels were examined using 2,7-dichlorodihydrofluorescein-diacetate and flow cytometry. The results revealed that cell proliferation was inhibited by lutein in a dose-dependent manner, and the apoptosis ratio gradually increased with lutein treatment under hypoxia as evident from flow cytometry-based analysis. Exposure to lutein inhibited hypoxia-mediated activation of HIF-1, NOTCH signaling and HES1 expression, and suppressed the hypoxia-induced expression of EMT-associated factors. Lutein markedly inhibited the invasion and migration of breast cancer cells under hypoxia. Hypoxia-induced production of ROS was also decreased by lutein. Furthermore, the ROS scavenger N-acetylcysteine also suppressed hypoxia inducible factor 1 and HES1 expression in breast cancer cells during hypoxia, but hydrogen peroxide (H2O2) levels were increased. Taken together, the results of the present study suggested that lutein may be a novel candidate for the chemoprevention of breast cancer. Furthermore, HES1 may be crucial in mediating the involvement of lutein in the suppression of hypoxia-driven ROS-induced breast cancer progression.