绿原酸对体外培养的人鼻咽癌细胞株CNE-1的作用

发布时间：2018-09-05 13:37

【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(P0.01)。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。
[Abstract]:Objective to investigate the effect of Lv Yuan acid (chlorogenic acid,CHA) on human nasopharyngeal carcinoma cell line CNE-1 in vitro and its mechanism. Methods Human nasopharyngeal carcinoma (NPC) cell line CNE-1, was resuscitated in aseptic condition for 4 passages. CHA, was added to the culture medium to make the final concentration of CHA up to 0.2550200500 渭 mol/L,. The best inhibitory concentration of CHA was screened by CCK-8 inhibition test in vitro. The optimal inhibitory concentration of CHA on CNE-1 cell line was selected. Telomerase activity, expression of tumor suppressor genes p27 and p16, expression of cyclin D1 and cyclin dependent kinase (CDK4) in human nasopharyngeal carcinoma cell line CNE-1 were detected. Results the optimal inhibitory concentration of CHA was 100 渭 mol/L. by CCK-8 inhibition test in vitro. The telomerase activity of human nasopharyngeal carcinoma cell line CNE-1 was 1.621 when the final concentration of CHA, was 100 渭 mol/L. The telomerase activity was significantly lower than that of the positive control and untreated CNE-1 cell line (P0.01). After treated with CHA for 10 days, the expression of tumor suppressor gene p27 and p16 was small, but almost no expression was found in the positive control group (HT1080 cells) and untreated CNE-1 cells. The expression of cyclin D1 and CDK4 in CNE-1 cells treated with CHA for 10 days was significantly lower than that in the positive control group (HT1080 cells) and untreated CNE-1 cells. Conclusion 100 渭 mol/L CHA for 10 days can decrease the telomerase activity and increase the expression of tumor suppressor gene in CNE-1 cell line. Reducing the expression of cyclin. Cha on human nasopharyngeal carcinoma cell line CNE-1 is achieved by activating the tumor suppressor gene and inhibiting the expression of cyclin.
【作者单位】：辽宁医学院病理生理学教研室;盘锦市中心医院耳鼻喉科;辽宁医学院解剖学教研室;
【基金】：辽宁省自然科学基金资助项目(No.2013022066,2013022048)~~
【分类号】：R739.63

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