表没食子儿茶素没食子酸酯(EGCG)对鼻咽癌细胞CNE1、CNE2放射增敏作用的初步研究  

The radiosensitization of EGCG on nasopharyngeal carcinoma( NPC) cell lines CNE1/CNE2 and its possible mechanism

作　　者：王红梅[1] 张伟军[1] 

机构地区：[1]广州医科大学附属肿瘤医院放疗一科,广东广州510095

出　　处：《现代肿瘤医学》2015年第14期1953-1958,共6页Journal of Modern Oncology

基　　金：广东省中医药局基金项目(编号:2010212);广东省科技厅基金项目(编号:20120318016)

摘　　要：目的:研究表没食子儿茶素没食子酸酯(EGCG)对鼻咽癌细胞CNE1、CEN2的放射增敏作用及其可能机制。方法:体外培养CNE1、CNE2细胞,CCK-8实验得到EGCG IC20值50μg/ml,即为实验浓度。实验分为4组,对照组:含有和其他组相等浓度的DMSO溶解剂;药物组:DMSO溶解的EGCG;照射组:X线照射组;实验组:EGCG联合X线照射组。体外培养CNE1、CNE2细胞至对数生长期,药物组及实验组分别给予药物即EGCG处理,培养24h后进行相应剂量的X线照射后收集细胞。采用克隆形成实验检测不同射线剂量(0、2、4、6、8、10Gy)处理后各组的克隆形成率、细胞存活率及放射增敏比(SER),流式细胞分析法检测各组细胞的凋亡情况及细胞周期,Western bolt检测Akt蛋白的表达,RT-PCR检测Akt mRNA的表达。结果:平板克隆实验:照射组及实验组随着X线剂量的逐步增加,克隆形成率随之下降,细胞存活分数下降,呈现剂量依赖效应(P<0.05);相同剂量下,实验组比照射组克隆形成率低,实验组Do、Dq明显低于单纯照射组(P<0.05),CNE1细胞株SER为1.4 9 6 2,CNE2细胞株SER为1.1 8 4 6,说明EGCG具有放射增敏作用(P<0.05)。流式细胞分析:各实验组与对照组相比,G2/M期百分比升高(P<0.05),表明存在G2/M期阻滞,单纯射线组比单纯药物组对G2/M期的阻滞作用更明显,二者联合的实验组表现为协同作用。实时荧光定量实验:各实验组与对照组相比,Akt mRNA表达下降(P<0.05),实验组比单纯照射组及单纯药物组更明显。Western blot实验:各实验组与对照组相比,Akt蛋白表达均有下降(P<0.05),实验组比单纯照射组及单纯药物组更明显。结论:EGCG对鼻咽癌细胞CNE1、CNE2具有放射增敏作用,其机制可能是通过抑制细胞增殖、促进细胞凋亡、抑制细胞周期、抑制射线诱导的Akt活化而产生。Objective：To study the radiosensitization of EGCG on nasopharyngeal carcinoma （NPC）cell lines CNE1 and CNE2 and possible mechanism.Methods：Cultured CNE1,CNE2 cells in vitro,CCK-8 experiment got EGCG IC20,50μg/ml,that was the experimental concentration.There were four groups,control group：Containing equal concentrations of DMSO dissolving agent with the other groups;Drug group：The EGCG dissolved by DMSO;Irradiated group：X-ray irradiation group;Experimental groups：EGCG combined X-ray irradiation.Cultured CNE1,CNE2 cells in vitro to logarithmic phase,the drug group and experimental group were given EGCG,fostered another 24 hours,irradiated cells with X-ray with appropriate dose.Through the tablet cloning experiments to detect the cloning efficiency,cell viability and radiosensitization ratio（SER） of each group were treated with appropriate dose X-ray （0,2,4,6,8,10Gy）.Flow cytometry was used to detect apoptosis and cell cycle,Western bolt detect the expression of Akt protein,RT-PCR to detect the expression of Akt mRNA.Results：Flat cloning experiments：Cloning efficiency and cell survival fraction decreased with increasing radiation dose in irradiated group and experimental group,appearing a dose-dependent effect （P ＜ 0.05）.The same dose group,the cloning efficiency of irradiation group was significantly lower than of the experimental group（P ＜0.05）,the same and Dq,the SER of CNE1 cell lines was 1.4962,CNE2 cell lines was 1.1846,that indicated that EGCG had radiosensitizing effect on nasopharyngeal carcinoma cells （P ＜ 0.05）.Flow cytometric analysis：Compared with the control group,the percentage of G2/M phase increased,indicating cells were arrested in G2/M phase,simple ray group was more obvious than the group of simple drug,the experimental group showed a synergistic effect（P ＜ 0.05）.RT-QPCR experiments：Compared with the control group,the down-regulation of Akt mRNA was significantly in the experimental group than the simple irradiation group and drug group（P

关 键 词：EGCG 鼻咽癌 Akt 放射增敏 

分 类 号：R739.6[医药卫生—肿瘤]