雷公藤甲素诱导鼻咽癌细胞凋亡作用  

Effect of triptolide on apoptosis of nasopharynx cancer

作　　者：王秀[1] 张竞竞[2] 张配[1] 孙小锦 刘哲[1] 张鑫宇[1] 刘浩[1] 

机构地区：[1]蚌埠医学院药学系,安徽省生化药物工程技术研究中心,安徽蚌埠233030 [2]蚌埠医学院第一附属医院肿瘤内科,安徽蚌埠233003

出　　处：《中国药理学通报》2014年第8期1147-1150,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81000992);蚌埠医学院科研项目(byky1346)

摘　　要：目的探讨雷公藤甲素诱导鼻咽癌细胞凋亡的作用及机制。方法 MTT测定雷公藤甲素对CNE-2Z细胞的抑制作用;碘化丙啶(propidium iodide,PI)染色测定雷公藤甲素对鼻咽癌细胞凋亡的影响;Western blot测定葡萄糖调节蛋白78(glucose regulated protein78,GRP-78)、Akt的表达及Akt的磷酸化水平;ROS荧光探针-二氢乙啶测定细胞内活性氧(reactive oxygen species,ROS)的水平。结果 MTT显示,雷公藤甲素对鼻咽癌细胞CNE-2Z的抑制作用随浓度增加及时间延长而增强;PI结果显示,雷公藤甲素能明显诱导鼻咽癌细胞的凋亡,25、50和100 nmol·L-1的雷公藤甲素作用细胞24 h,CNE-2Z细胞的凋亡率分别为14.0%、26.9%和34.4%;Western blot显示,雷公藤甲素对CNE-2Z细胞GRP-78表达无明显影响,但可减弱Akt的表达及磷酸化水平;氧化应激实验结果表明,雷公藤甲素作用4 h后,能增加鼻咽癌细胞CNE-2Z内ROS的水平。结论雷公藤甲素具有抑制鼻咽癌细胞CNE-2Z增殖的作用,且具有浓度与时间依赖性。其机制可能是雷公藤甲素通过诱导氧化应激,抑制细胞内Akt的表达及其磷酸化,调节下游的信号通路,进而促进CNE-2Z细胞的凋亡。Aim Toexploretheinhibitioneffectof triptolide on nasopharynx cancer, and the mechanism. Methods Theinhibitionofcellproliferationwasde-tected by MTT assay;the cell apoptosis was analyzed by flow cytometry with propidium iodide staining. The ex-pressions of glucose regulated protein 78 （ GRP-78 ） , Akt and pAkt in cells were examined by Western blot;the effect of triptolide on reactive oxygen species （ ROS） accumulation was detected by ROS Fluorescent Probe-DHE.Results MTTassayshowedthatthe growth of nasopharynx cancer was inhibited by triptol-ide , and the inhibition occurred in a dose and time-de-pendent manner following triptolide treatment in CNE-2Z nasopharynx cancer cells. Propidium iodide staining revealed that the apoptosis of CNE-2 Z cells was in-duced remarkably by triptolide. After CNE-2Z cells treated with 25, 50,100 nmol·L-1 of triptolide for 24 h, the apoptosis rate was 14%,26. 9% and 34. 4% re-spectively. Western blot experiment showed that the expression of GRP-78 had no significant change follow-ing triptolide treatment in CNE-2 Z nasopharynx cancer cells for 24 h, but the expression and the phosphoryla-tion level of Akt were strikingly decreased. The experi-ment of ROS uncovered that CNE-2 Z nasopharynx cancer cells increased generation of ROS after treat-ment with triptolide for 4 hours, and acted cells in a dosedependentmanner.Conclusions Triptolidecan inhibit the growth of CNE-2 Z nasopharynx cancer cells in a dose and time-dependent maner. The mechanism may be related with the point that triptolide can induce oxidative stress, incease ROS, inhibit the expression and the phosphorylation level of Akt,then promote the apoptosis of CNE-2Z cells.

关 键 词：雷公藤甲素 鼻咽癌细胞 氧化应激 活性氧 凋亡 

分 类 号：R284.1[医药卫生—中药学]