蛋白酶体抑制剂对甲状腺未分化癌细胞ATF-4表达影响的研究  

Effects of proteasome inhibitors on expression of activating transcription factor-4 of undifferentiated thyroid cancer cells

作　　者：张海燕[1] 吕岩[2] 孟欣[3] 都镇先[4] 邓娓娓[1] 王华芹[3] 

机构地区：[1]中国医科大学附属第一医院老年病科,辽宁沈阳110001 [2]中国医科大学护理学院,辽宁沈阳110001 [3]中国医科大学基础医学院生化与分子生物教研室,辽宁沈阳110001 [4]中国医科大学附属第一医院内分泌科,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2012年第16期1212-1215,共4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)

摘　　要：目的:探讨蛋白酶体抑制剂对甲状腺未分化癌FRO细胞中活化转录因子4(ATF-4)表达的影响,以及ATF-4在蛋白酶体抑制剂诱导FRO细胞凋亡中的作用。方法:选取人甲状腺未分化癌细胞系FRO,分别设空白对照组和蛋白酶体抑制剂MG132处理组;利用微小RNA干扰技术,减少ATF-4的表达,实时定量PCR法、蛋白质印迹法分别检测各组细胞中ATF-4、氧调节蛋白150(ORP150)mRNA和蛋白表达;流式细胞仪(FCM)检测细胞凋亡。结果:与空白对照组相比,蛋白酶体抑制剂MG132显著增加FRO细胞系中ATF-4基因表达水平,P<0.01;与空白对照组和随机序列核酸siRNA组相比,siATF-4处理组中ATF-4、ORP150mRNA及蛋白表达水平显著降低(P<0.01),其凋亡率显著增加,P<0.01。结论:蛋白酶体抑制剂能够上调甲状腺未分化癌FRO细胞系中ATF-4基因的表达水平,siATF-4具有降低ATF-4及OPR150基因表达的作用,同时增加蛋白酶体抑制剂诱导甲状腺未分化癌FRO细胞的凋亡作用。OBJECTIVE： To investigate the role of activating transcription factor （ATF）-4 in thyroid cancer ceil death induced by proteasome inhibitors. METHODS： The undifferentiated thyroideaneer FRO cells were treated with vehicle, proteasome inhibitor MG132; ATF-4 expression levels were knockdown by RNA interference; ATF-4 and 150 × 10^3 oxygen-regulated protein （ORP150） mRNA and protein levels were analyzed by using real time RT-PCR and Western blot,respectively; the apoptotie rate was analyzed by using flow cytometry（FCM）. RESULTS： MG132 significantly in- creased the mRNA and protein levels of ATF-4 （P〈0.01） in undifferentiated thyroid cancer FRO cells small interfering RNA against ATF-4 （si ATF-4） markedly reduced the ATF-4 and ORP150 mRNA and protein levels when compared to vehicle or scramble siRNA （P〈0. 01） ,in addition siATF-4 dramatically increased MG132 mediated increase in apoptotic cells （P〈0.01）. CONCLUSION： Proteasome inhibitors increase the ATF-4 expression in undifferentiated thyroid cancer FRO cells and the suppression of ATF-4 and ORP150 expression by siATF-4 significantly enhances the apoptosis of FRO cells induced by proteasome inhibitors.

关 键 词：甲状腺肿瘤/病理学 细胞凋亡 蛋白酶体抑制剂 活化转录因子4 

分 类 号：R736.1[医药卫生—肿瘤]