白藜芦醇阻断信号通路对鼻咽癌细胞生长的影响

Effect of Resveratrol on blocking growth of nasopharyngeal carcinoma cells

摘要:

目的 通过体内和体外研究探讨白藜芦醇在人核孔复合物细胞（NPC）中的抗癌活性。方法 用 MTT 实验检测白藜芦醇对CNE-1 细胞和CNE-2Z 细胞（两种具有不同分化程度的NPC 细胞株）生长增殖 的影响。用流式细胞仪检测白藜芦醇诱导NPC 细胞凋亡情况和NPC 细胞周期分布变化。Western blot 检测 涉及细胞凋亡调控的数个重要基因和与细胞周期调控有关的数种Cyclins 的蛋白表达水平。在裸鼠体内进行 CNE-2Z 细胞移植瘤实验。结果 用白藜芦醇处理NPC 细胞后，白藜芦醇不仅以时间和剂量依赖的方式降低 NPC 细胞的增殖能力，还可以剂量依赖的方式诱导NPC 细胞的凋亡。流式细胞仪分析结果表明，白藜芦醇处 理可将NPC 细胞生长阻滞于S- 期和G2/M- 期。白藜芦醇处理可下调Bcl-2 和低氧诱导因子1（HIF-1）蛋白 的表达，上调Caspase-3 蛋白表达。此外，白藜芦醇处理不仅可以降低蛋白激酶B（Akt）、p70 核糖体S6 蛋白激 酶（p70S6K）和真核翻译起始因子4E- 结合蛋白1（4E-BP-1）的磷酸化水平，也可降低参与细胞周期调控的 数个细胞周期蛋白（Cyclins）的表达水平。白藜芦醇可抑制裸鼠体内NPC 移植瘤的生长，进一步确认白藜芦醇 对NPC 生长的治疗效果。结论 在人NPC 细胞株中，白藜芦醇可能是通过干扰Akt/p70S6K 信号通路而发挥有 效的抗增殖和促凋亡作用。

Abstract:

Objective To explore anticancer activity of Resveratrol in human nuclear pore complex cells (NPC). Methods MTT test was utilized to measure cellular growth of CNE-1 and CNE-2Z. Flow cytometry was performed to examine apoptosis and cell cycle status. Expression of several apoptosis associated proteins and cyclins was measured by Western blot. CNE-2Z xenografts on nude mice were performed to further validate anticancer capability of Resveratrol in vivo. Results Cell proliferation of NPC cells was inhibited by Resveratrol in timedependent and dose-dependent manners, while apoptosis was significantly induced after being exposed to Resveratrol in dose-dependent manner. Flow cytometry results showed that Resveratrol arrestted NPC cells at the S-phases and G2/M- phases. Expression of Bcl-2 and hypoxia-inducible factor-1a (HIF-1α) were downregulated while caspase-3 as well as several cyclins involved in cell cycle regulation were upregulated with treatment of Resveratrol. Moreover, Resveratrol significantly decreased the phosphorylation levels of Akt, p70S6K and eukaryotic translation initiation factor 4E-binding protein-1 (4E-BP-1). In vivo study showed that Resveratrol dramatically inhibited the growth of NPC tumor xenografts. Conclusions Collectively, our findings suggest that Resveratrol exerts anti-proliferative and pro-apoptotic effects on human NPC cells through interfering with the Akt/p70S6K signaling pathways.