佛手柑内酯对鼻咽癌细胞周期的影响  

Effect of Bergapten on Cells Cycle in Nasopharyngeal Carcinoma

作　　者：林碧华[1] 万树伟[1] 刘付梅[1] 崔仲宜 张鑫[1] 李祥勇[1] 

机构地区：[1]广东医学院生物化学与分子生物学教研室,广东东莞523000

出　　处：《中国药学杂志》2014年第10期837-842,共6页Chinese Pharmaceutical Journal

基　　金：东莞市高等院校科技计划一般项目(2011108102044);湛江市科技攻关项目(2011C3108012;2013B01091);广东医学院博士启动基金(B2011022);广东医学院青年基金(Q2012005)

摘　　要：目的探讨佛手柑内酯对鼻咽癌细胞周期的影响及可能的作用机制。方法利用CCK-8法测定佛手柑内酯对鼻咽癌细胞CNE-2和HONE-1的体外抑制活性,并绘制1、10、100μmol·L-13个浓度的佛手柑内酯对2种鼻咽癌细胞作用后的生长曲线,然后利用PI染料法检测各组细胞的周期分布。利用Western blotting检测CDK4、CDK6、CDK2、Cyclin D1和Cyclin E2的蛋白表达量,并检测各组β-catenin、GSK-3β及GSK-3β的S9位点蛋白磷酸化水平。结果佛手柑内酯对鼻咽癌细胞CNE-2和HONE-1具有较显著的体外抑制活性,呈明显的浓度依赖性(CNE-2:r=0.926,P<0.05;HONE-1:r=0.959,P<0.05)及时间依赖性(CNE-2:r=0.991,P<0.05;HONE-1:r=0.963,P<0.05),随着佛手柑内酯浓度的增加,两鼻咽癌细胞处于G0-G1期的细胞数目比例上升,S期与G2-M的比例下降。相对于对照组,佛手柑内酯能显著降低实验组细胞CDK4、CDK6、CDK2、Cyclin D1、Cyclin E2和β-catenin的蛋白表达量,但对GSK-3β的蛋白表达水平无显著影响,对GSK-3β的S9位点蛋白磷酸化水平的影响也无明显规律。结论佛手柑内酯能通过阻滞鼻咽癌细胞周期运行抑制其体外增殖能力,这一过程可能与部分周期蛋白的下调及wnt/β-catenin信号通路的被抑制有关。OBJECTIVE To investigate the blocking effect of bergapten on cell cycle in nasopharyngeal carcinoma（ NPC） cells. METHODS The inhibition was tested by CCK-8 assay after the NPC cells（ CNE-2 and HONE-1） being exposed to bergapten for 48 h. The bergapten concentrations were as follows： 1,10 and 100 μmol·L- 1. The cell cycle was assayed by PI staining and following flow cytometry analysis,and the CDK4,CDK6,CDK2,Cyclin D1,Cyclin E2,β-catenin,GSK-3β and p-GSK-3β（ S9） protein levels were detected by Western blotting. RESULTS Bergapten was able to significantly inhibit CNE-2 and HONE-1 NPC cancer cell proliferations,and the effect was dose-（ CNE-2： r = 0. 926,P 0. 05; HONE-1： r = 0. 959,P 0. 05） and time-dependent（ CNE-2： r = 0. 991,P 0. 05; HONE-1： r = 0. 963,P 0. 05）. Drug treatment induced a block in the G0/ G1phase and decreased protein levels of CDK4,CDK6,CDK2,Cyclin D1,Cyclin E2 and β-catenin,but the protein levels of GSK-3β and the phosphorylation of p-GSK-3β（ S9） didn＇t change. CONCLUSION Bergapten can block cells cycle and inhibit NPC cells＇ proliferations,and those effects might be related to suppressing of cells cycle-related protein and interfering with the wnt / β-catenin signal pathway.

关 键 词：佛手柑内酯 鼻咽癌 细胞周期 Β-CATENIN GSK-3Β 

分 类 号：R965[医药卫生—药理学]