曲古抑菌素对肿瘤坏死因子α诱导U937细胞IκB-α蛋白降解的影响  

Effect of trichostatin A on degradation of IκB-α induced by TNF-α in U937 cells

作　　者：崔国惠[1] 陈卫华[1] 陈燕[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院,湖北武汉430022

出　　处：《中国医院药学杂志》2007年第3期285-288,共4页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金(编号:30472267)

摘　　要：目的:探讨组蛋白去乙酰化酶抑制剂曲古抑菌素(trichostatin A,TSA)与肿瘤坏死因子α(TNF-α)分别或联合作用于人类单核细胞白血病细胞株U937细胞前后,IκB-α蛋白的表达变化及其在NF-κB激活过程中的作用。方法:分别将TSA、TNF-α单独或联合作用于U937细胞,用免疫印迹法及荧光显微镜检测NF-κB/P65蛋白在该细胞中的表达及定位的变化;流式细胞术和免疫印迹法分析IκB-α蛋白表达的变化。结果:①荧光显微镜观察:分别作用45min后,NF-κB/P65蛋白分布于对照组及TSA处理组的细胞浆;而TNF-α处理组胞浆、胞核中均可见P65蛋白分布;TSA+TNF-α联合处理组仅胞浆表达P65蛋白。免疫印迹结果同样证实,作用相同时间后,TNF-α处理组细胞核表达P65蛋白,TSA+TNF-α联合处理组细胞核内P65蛋白表达较前者明显减少;②流式细胞术结果表明,经相同时间处理后,TSA+TNF-α联合处理组的IκB-α蛋白平均荧光强度较TNF-α处理组明显增高(P<0.05);免疫印迹结果也显示,作用45min后,TNF-α处理组细胞IκB-α蛋白表达较对照组明显降低,TSA+TNF-α联合处理组细胞IκB-α的表达较前者明显增高,但在作用2h后二者IκB-α蛋白表达差异无显著性。结论:TSA能部分抑制TNF-α诱导的U937细胞IκB-α的降解,从而抑制NF-κB的激活。TSA有可能通过影响NF-κB信号途径发挥抗肿瘤作用。OBJECTIVE To investigate the effect of trichostatin A,one kind of HDAC inhibitors,on expression of IκB-α induced by TNF-α and activation of NF-κB in U937 cells. METHODS U937 cells were treated with TSA,TNF-α respectively or with both, the protein expression and subcellular localization of NF-κB/p65 were observed by western blot and fluorescence microscopy respectively; the expression and degradation of IκB-α were investigated through by flow cytometry and western blot. RESULTS （1）Under fluorescence microscopy,NF-κB/p65 was only localized in cytoplasm of empty control and group -treated with TSA for 45 min, p65 was localized in nucleus of U937 cells treated with TSA and TNF-α, while it was expressed in both cytoplasm and nucleus of cells treated with TNF-α. Western blot also showed the same result ：TNF-α-treated group cells nuclears expressed much more p65 protein than those treated with TNF-α plus TSA for 45 min. （2）After being treated with TSA and TNF-α for 45 min, the protein expression of IκB-α in U937 cells detected by flow cytometry was stronger than those only treated with TNF-α （P〈0. 05）. Through Western blot, it was found that IκB-α expression was fewer in TNF-α-treated group compared to control; which was obviously enhanced in group treated with TSA plus TNF-α than that treated with TNF-α for 45 min,but it was no differences between them after being treated for 2 h. CONCLUSION TSA can partially inhibit degradation of IκB-α induced by TNF-α and NF-κB activation in U937 cells. TSA can work through NF-κB signal pathway against tumor,which can be a target of TSA for cancer therapy.

关 键 词：TSA 肿瘤坏死因子Α NF-ΚB/P65 IΚB-Α U937细胞株 

分 类 号：R979.1[医药卫生—药品]