Grape seed extract triggers apoptosis in Caco-2 human colon cancer cells through reactive oxygen species and calcium increase: extracellular signal-regulated kinase involvement
Grape seed extract triggers apoptosis in Caco-2 human colon cancer cells through reactive oxygen species and calcium increase: extracellular signal-regulated kinase involvement
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DOI：
10.1017/s0007114512006095
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发表时间：
2013-09-14
影响因子：
3.6
通讯作者：
Bizzarri, Mariano
中科院分区：
文献类型：
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作者：
Dinicola, Simona;Mariggio, Maria Addolorata;Bizzarri, Mariano
来自 Italia、Palieri 和 Red Globe 品种的葡萄籽提取物 (GSE) 以剂量依赖性方式抑制 Caco-2 人结肠癌细胞的生长并诱导细胞凋亡。为了研究支持细胞凋亡过程的机制，我们分析了活性氧 (ROS) 的产生、细胞内 Ca2+ 处理和细胞外信号调节激酶 (ERK) 激活。暴露于 GSE 后，Caco-2 细胞中的 ROS 和细胞内 Ca2+ 水平增加，同时 ERK 失活。由于 ERK 活性被认为对于促进生存途径至关重要，因此抑制该激酶可能在 GSE 介导的抗癌作用中发挥相关作用。事实上，用 ROS 清除剂 N-乙酰半胱氨酸进行预处理，可以逆转 GSE 诱导的细胞凋亡，并促进 ERK 磷酸化。乙二醇四乙酸介导的细胞外 Ca2+ 内流抑制作用增强了这种作用。 ROS 和 Ca2+ 流入抑制反过来又增加了 ERK 磷酸化，因此几乎完全抑制了 GSE 介导的细胞凋亡。这些数据表明，GSE 触发了一种以前未被识别的基于 ERK 的机制，涉及 ROS 产生和细胞内 Ca2+ 增加，最终导致癌细胞凋亡。
Grape seed extract (GSE) from Italia, Palieri and Red Globe cultivars inhibits cell growth and induces apoptosis in Caco-2 human colon cancer cells in a dose-dependent manner. In order to investigate the mechanism(s) supporting the apoptotic process, we analysed reactive oxygen species (ROS) production, intracellular Ca2+ handling and extracellular signal-regulated kinase (ERK) activation. Upon exposure to GSE, ROS and intracellular Ca2+ levels increased in Caco-2 cells, concomitantly with ERK inactivation. As ERK activity is thought to be essential for promoting survival pathways, inhibition of this kinase is likely to play a relevant role in GSE-mediated anticancer effects. Indeed, pretreatment with N-acetyl cysteine, a ROS scavenger, reversed GSE-induced apoptosis, and promoted ERK phosphorylation. This effect was strengthened by ethylene glycol tetraacetic acid-mediated inhibition of extracellular Ca2+ influx. ROS and Ca2+ influx inhibition, in turn, increased ERK phosphorylation, and hence almost entirely suppressed GSE-mediated apoptosis. These data suggested that GSE triggers a previously unrecognised ERK-based mechanism, involving both ROS production and intracellular Ca2+ increase, eventually leading to apoptosis in cancer cells.