蛋白酶体抑制剂YSY01A不依赖NF-κB通路抑制乳腺癌细胞增殖及诱导凋亡研究  

YSY01A as the NF-κB-independent proteasome inhibitor inhibits proliferation and induces apoptosis in breast cancer cells

作　　者：张梅[1,2] 袁霞[1] 徐波[1] 冉福香[1] 吴军[1] 葛泽梅[3] 李润涛[3] 崔景荣[1] 

机构地区：[1]北京大学药学院天然药物及仿生药物国家重点实验室,新疆石河子832000 [2]新疆石河子大学药学院药理系,新疆石河子832000 [3]北京大学药学院化学生物学系,北京100191新疆石河子832000

出　　处：《中国药理学通报》2014年第3期34-39,共6页Chinese Pharmacological Bulletin

基　　金：国家科技部"重大新药创制"科技专项基金(No2009ZX0930010);国家自然科学基金资助项目(No81172915)

摘　　要：目的研究YSY01A对乳腺癌的增殖抑制作用及其抗癌机制。方法荧光底物法检测蛋白酶体活性;蛋白酶体荧光探针法研究YSY01A与蛋白酶体β催化亚基结合;Annexin V-FITC、JC-1探针检测YSY01A对细胞凋亡和线粒体膜电位的影响;用蛋白免疫印迹法检测细胞内的蛋白表达。结果YSY01A抑制细胞内外蛋白酶体的活性,其中对CT-L的抑制作用最强,对T-L的作用比PS341强3-4倍;MCF-7对YSY01A最敏感,且比PS341敏感性高;YSY01A以时间依赖性诱导细胞凋亡,与对照组比较,差异均有显著性(P<0.05)。YSY01A增加ubiquitin、P-IκBα、wt-p53、Bax、endonuclearse G蛋白表达,减少IκBα、Bcl-xL、XIAP水平,不影响NF-κB表达。结论 YSY01A是新蛋白酶体抑制剂,具有较强的抑制MCF-7细胞增殖的作用,抗癌机制与诱导细胞凋亡有关。Aim To investigate proliferative inhibition and the mechanisms of YSY01A in breast cancer cells. Methods Three special fluorogenie substrates were used in proteasome activity test. A proteasome probe was used to analyze expression of active subunits after proteins were resolved in 12.5% SDS-PAGE gel. The inhibition of cells proliferation was tested by SRB. Ap- optotie cells were determined by FCM with annexin-V, and the potential of mitoehondria membrane was measured with JC-1 probe. Expression of proteins was ana- lyzed by Western blot. Results YSY01A inhibited three catalytic sites of proteasome; the inhibitory effect of YSY01A on T-L was stronger than PS341 by 3M folds, the inhibitory effects on other active sites were somewhat weak. YSY01A reduced proliferations of three types of breast cancer cells, but MCF-7 cells were highly sensitive. YSY01A induced MCF-7 cells apoptosis and lessened mitoehondrial membrane poten- tial. Although YSY01A did not change NF-KB level, it obviously enhanced levels of ubiquitin proteins, Bax, puma, endonuelease G, P-IKBoL and wt-p53 proteins, decreased Bel-xL and activated PARP in a time-de- pendent manner. Conclusion YSYO1A, as the NF- KB-independent proteasome inhibitor, inhibits breast cancer cells growth by inducing apoptosis.

关 键 词：蛋白酶体 抑制剂 增殖 凋亡 NF-ΚB 乳腺癌 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]