摘要
目的:探讨氧化应激在启动蛋白酶体抑制剂诱导甲状腺癌细胞凋亡中的作用。方法:选取4种人甲状腺未分化癌细胞系ARO、FRO、KTC2和8305C,分别设空白对照组、万珂处理组和万珂+钛剂联合组;流式细胞仪(FCM)检测细胞凋亡情况;Caspase-3活性测试法检测各组细胞Caspase-3活性;用荧光分光光度计法检测各组细胞蛋白酶体活性和细胞内ROS水平。结果:与空白对照组相比,蛋白酶体抑制剂万珂对人甲状腺癌细胞系FRO和KTC2具有很强的细胞凋亡诱导作用;使FRO和KTC2细胞中Caspase-3的活性显著增高,P<0.01;可持续快速抑制蛋白酶体活性,但在4组细胞中差异无统计学意义,P>0.05;万珂可显著提高活性氧簇在FRO和KTC2甲状腺癌细胞中的表达。万珂组与万珂+钛剂组活性氧簇水平在4种甲状腺癌细胞中差异均有统计学意义,P<0.05;在FRO和KTC2甲状腺癌细胞中,万珂组与万珂+钛剂组中的细胞凋亡率差异有统计学意义,P<0.01。结论:蛋白酶体抑制剂可以提高活性氧簇在甲状腺未分化癌细胞中的表达,促进肿瘤细胞的凋亡,并且这种效应可以被抗氧化剂所抑制。
OBJECTIVE： To investigate the role of reactive oxygen species（ROS） in the proteasome inhibition-induced apoptosis of thyroid cancer cells.METHODS： ARO,FRO,KTC2 and 8305C human undifferentiated thyroid cancer cells were treated with vehicle,bortezomib alone or bortezomib in combination with tiron;Flowcytometry was used to measure apoptotic cells;The activity of caspase was measured using spectrometry;The activity of proteasomes and intracellular ROS levels were measured using fluorospectrophotometry.RESULTS： Compared with vehicle,proteasome inhibitor botezomib significantly induced apoptosis in FRO and KTC2 thyroid cancer cells;Activities of caspase-3 in FRO and KTC2 cells were significantly increased by botezomib（P0.01）;Botezomib rapidly and continuously decreased the activities of proteasomes,but had no statistical difference in 4 different thyroid cancer cell lines（P0.05）;Botezomib significantly enhanced the intracellular levels of ROS in FRO and KTC2 thyroid cancer cells.The intracellular ROS levels were significantly different in botezomb-treated cells when compared with botezomb and tiron-treated cells（P0.05）;Tiron significantly blocked apoptosis of FRO and KTC2 cells induced by botezomib（P0.01）.CONCLUSIONS： Proteasome inhibitor can induce apoptosis of thyroid cancer cells via promotion of ROS generation.Reductants could suppress apoptosis induced by proteasome inhibitor.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2012年第4期263-267,共5页
Chinese Journal of Cancer Prevention and Treatment
基金
辽宁省科技攻关项目(2010225032)
辽宁省教育厅资助项目(2009A765)
中国医科大学附属第一医院科学研究基金(fsfh 1008)
