萝卜硫素介导Akt/p70S6K信号传导诱导人鼻咽癌CNE-2细胞凋亡

Effects of sulforaphane-induced Akt/p70S6K signal transduction pathway on apoptosis of human nasopharyngeal cancer cells CNE-2

目的:研究萝卜硫素对人鼻咽癌CNE-2细胞增殖、凋亡的影响及其对Akt/p70S6K信号传导的作用。方法:采用CCK-8实验分析萝卜硫素对CNE-2细胞生长增殖的影响,流式细胞仪检测萝卜硫素对CNE-2细胞凋亡率及细胞周期分布的影响,Western blot技术检测CNE-2细胞中调控细胞周期的相关蛋白及Akt/p70S6K信号通路关键蛋白的表达水平。结果:采用萝卜硫素干预CNE-2细胞后,细胞增殖抑制率及凋亡率相对于对照组呈浓度依赖的方式显著增高,差异有统计学意义( P < 0.05 );流式细胞仪分析提示,萝卜硫素干预能够将细胞停留在S期及G 2/M期,并且萝卜硫素还能抑制细胞周期蛋白Cyclin A、Cyclin B、Cyclin D、Cyclin E以及CDK/p34的表达水平,与对照组相比差异有统计学意义( P < 0.05 );此外,萝卜硫素还能抑制Akt/p70S6K信号通路关键蛋白p-Akt及p70S6K蛋白表达水平( P < 0.05 ),但萝卜硫素与Akt激动剂IGF-I共同处理细胞后,上述蛋白表达量与对照组相比没有统计学差异( P > 0.05 )。结论:萝卜硫素可能通过干扰CNE-2细胞中Akt/p70S6K信号传导而发挥有效的抗增殖及促凋亡的作用。

AIM : To study the effect of sulforaphane on the proliferation and apoptosis of CNE-2 cells, and to investigate the role of sulforaphane in the Akt/p70S6K signaling pathway. METHODS : CCK-8 test was used to detect cellular growth of CNE-2. Flow cytometry was carried out to observe apoptosis and cell cycle status. Expression levels of several cell cycle associated proteins and Akt/p70S6K pathway related proteins were determined by Western blot. RESULTS :Cell proliferation inhibitory rates and apoptotic rates of CNE-2 cells in various doses of sulforaphane groups were significantly increased as compared with control group with a dose-dependent manner ( P < 0.05 ). Flow cytometry results showed that sulforaphane arrestted NPC cells at the S-phases and G 2/M-phases, and the expression levels of Cyclin A, Cyclin B, Cyclin D, Cyclin E and CDK/p34 were greatly decreased in sulforaphane group as compared with control group ( P < 0.05 ). Moreover, the expression levels of p-Akt and p70S6K in sulforaphane group were also decreased as compared with control group ( P < 0.05 ). However, the expression levels of these proteins above in sulforaphane+ IGF-I group unchanged as compared with control group ( P > 0.05 ). CONCLUSION : Sulforaphane exerts anti-proliferative and pro-apoptotic effects on CNE-2 cells through interfering with the Akt/p70S6K signaling pathways.