摘要
目的探讨重组人内皮抑素(rh-endostatin,rh-ES)对鼻咽癌CNE2细胞增殖、凋亡的影响。方法分别以25、50、100、200、400μg/ml的rh-ES体外处理人鼻咽癌细胞株CNE2,同时设空白对照组和细胞对照组。利用四甲基偶氮唑蓝法(MTT)检测不同浓度和时间rh-ES作用下的细胞增殖状态;流式细胞仪检测细胞周期及凋亡;透射电镜观察CNE2细胞超微结构的变化。结果①MTT检测显示rh-ES(25～400μg/ml)能抑制CNE2细胞的增殖,不同浓度的rh-ES对人鼻咽癌细胞株CNE2具有不同程度的生长抑制作用,且存在时间剂量效应,以400μg/ml rh-ES作用72 h后抑制效果最为显著,各浓度组相比差异具有统计学意义。②流式细胞仪检测结果发现与CNE2细胞对照组相比,rh-ES实验组处于细胞周期G0/G1期的细胞明显增多,细胞凋亡率增加(P<0.05)。③电镜观察,rh-ES实验组CNE2细胞出现凋亡特征,细胞和细胞器皱缩,核染色质边集碎裂,核膜消失。结论 rh-ES能显著抑制鼻咽癌细胞CNE2细胞的增殖,并具有时间-剂量依赖性,其主要机制可能为诱导细胞凋亡,调整细胞周期分布。
Objective To evaluate the inhibitory effect and the possible mechanism of rh-endostatin （rh-ES） on nasopharyngeal carcinoma cells （CNE2）. Methods CNE2 cells were treated with rh-ES of different concentrations （25, 50, 100, 200 and 400 μg /ml, respectively ） in vitro. Methyl thiazolyl tetrazolium （MTT） assay was used for detecting the cell proliferation in rh-ES treatment at different concentrations and different time intervals. Flow cytometry was used for detecting the effect of rh-ES on cell cycle and apoptosis. Transmission electron microscope was used for observing the cellular morphology of CNE2 cells. Results （1） MTT detection showed that rh-ES significantly inhibited the growth of nasopharyngeal carcinoma cell CNE2 in both time- and dose-dependent manners. 72 hours after rh-ES treatment it was at 400 p,g/ml, and the inhibitory rate was 41.89% , which was higher than those of the other groups. （2） Flow cytometry showed that the proportion of CNE2 cells arrested in G0/G1 phase and cell apoptosis rate after rh-ES treatment were significantly higher than those of the control group respectively （ P 〈 0.05 ）. （3） Transmission electron microscope identified the characteristics of apoptosis appeared in CNE2 cell of rh-ES group, such as crenation of cell organ, chips of chromatin margination and disappearance of caryotheca. Conclusion By inducing apoptosis and changing the cell cycle distribution, rh-ES can inhibit the proliferation of nasopharyngeal carcinoma cell CNE2 in both time- and dose-dependent manners.
出处
《中国耳鼻咽喉颅底外科杂志》
CAS
2012年第5期345-348,353,共5页
Chinese Journal of Otorhinolaryngology-skull Base Surgery
基金
云南省应用基础研究面上项目(2007C254M)
关键词
重组人内皮抑素
鼻咽癌
增殖
凋亡
rh-endostatin
Nasopharyngeal neoplasm
Proliferation
Apoptosis
作者简介
马静，男，主治医师。
通讯作者：李晓江，Email：xiaDjiangle@yahoo．com．cn．
