绿原酸对体外培养的人鼻咽癌细胞株CNE-1的作用  

Effects of chlorogenic acid on cultured human nasopharyngeal carcinoma cell line CNE-1 in vitro

作　　者：姚素艳[1] 李全胜 郑德宇[3] 

机构地区：[1]辽宁医学院病理生理学教研室,辽宁锦州121001 [2]盘锦市中心医院耳鼻喉科,辽宁盘锦124000 [3]辽宁医学院解剖学教研室,辽宁锦州121001

出　　处：《西安交通大学学报（医学版）》2014年第6期837-842,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：辽宁省自然科学基金资助项目(No.2013022066;2013022048)~~

摘　　要：目的探讨绿原酸(chlorogenic acid,CHA)对体外培养状态下人鼻咽癌细胞株CNE-1的作用及其机制。方法在无菌状态下复苏人鼻咽癌细胞株CNE-1,取传4代的细胞,向培养基内加入CHA,使其终浓度达到0、25、50、100、200、500μmol/L,应用CCK-8体外抑制实验进行筛选CHA的最佳抑癌浓度。选择CHA最佳抑癌浓度,作用于CNE-1细胞系。检测人鼻咽癌细胞株CNE-1中端粒酶活性、抑癌基因p27和p16的表达、细胞周期蛋白(cyclin D1)和细胞周期蛋白依赖激酶(CDK4)的表达。结果经CCK-8体外抑制实验进行筛选CHA的最佳抑癌浓度为100μmol/L。人鼻咽癌细胞株CNE-1的培养基中加入终浓度为100μmol/L的CHA,维持10d,CNE-1细胞株中其端粒酶活性为1.621,远低于试剂盒中的阳性对照和未处理的CNE-1细胞株,差异具有统计学意义(P<0.01)。应用CHA处理CNE-1细胞株10d后,抑癌基因p27和p16均有少量的表达,而在阳性对照组(HT1080细胞)和未处理的CNE-1细胞几乎没有表达。应用CHA处理CNE-1细胞株10d后,cyclin D1和CDK4的表达量与阳性对照组(HT1080细胞)和未处理的CNE-1细胞相比明显降低。结论应用100μmol/L CHA 10d,可以降低CNE-1细胞株端粒酶活性、增加抑癌基因的表达,降低细胞周期蛋白的表达。CHA对人鼻咽癌细胞株CNE-1的作用是通过活化抑癌基因和抑制细胞周期蛋白的表达来实现的。Objective To explore the effects and mechanisms of chlorogenic acid （CHA） on the human nasopharyngeal carcinoma cell line CNE-1 cultured in vitro. Methods The human nasopharyngeal carcinoma cell line CNE-1 underwent resuscitation in sterile conditions. CHA was added into the CNE-1 culture media at the concentrations of 0, 25, 50, 100, 200 and 500 μmol/L for 7 days. The optimal concentration of CHA which inhibited the CNE-1 was determined by CCK8 inhibition experiment in vitro. After CNE-1 was cultured at the optimal concentration of CHA for 10 days, telomerase activation was detected by ELISA method. The expressions of anti-oncogene p27 and p16 as well as cyclin D1 and CDK4 were determined by Western blot. Results The optimal concentration of CHA for inhibiting CNE-1 cell line was 100 μmol/L determined by CCK8 inhibition experiment. Telomerase activation of CNE-1 was decreased to 1. 621 after CNE-1 was treated with 100 μmol/L CHA for 10 days. The activation was significantly lower than that in the positive group and the normal cultured CNE-1 group （P〈0.01）. Anti-oncogene p27 and p16 were expressed in a low quantity after CNE-1 was treated with 100 t^mol/L CHA for 10 days. But there was hardly any p27 or p16 expression in the positive group （HT1080） and the normal cultured CNE-I group （CHA concentration being 0 μmol/L） determined by Western blot method. Theexpressions of cyclin D1 and cyclin dependence kinase 4 （CDK4） were lower after CNE-1 received 100 μmol/L CHA for 1O days than those in the HT1080 cell line and the normal cultured CNE-1 （CHA concentration being 0 μmol/L）. Conclusion CHA has the functions of decreasing telomerase activation of CNE-1 cell line, increasing the anti- oncogene expression and decreasing cyclin D1 expression. The inhibitory function of CHA on CNE-1 cell line is realized by activating the expression of anti-oncogene and inhibiting the expression of cyclin D1.

关 键 词：绿原酸 鼻咽癌 抑癌基因 细胞周期蛋白 端粒酶 P27 P16 

分 类 号：R766.3[医药卫生—耳鼻咽喉科]