蛋白酶体抑制剂PS-341诱导U266细胞凋亡与胞浆内[Ca^(2+)]变化的关系  

Relationship of Proteasome Inhibitor PS-341 Induced Apoptosis with Cytoplasmic [Ca^(2+)] Changes in U266 Cells

作　　者：林如峰[1] 陆化[1] 刘澎[1] 王永韧[1] 沈文怡[1] 吴雨洁[1] 张建富[1] 葛峥[1] 汪承亚[1] 李建勇[1] 

机构地区：[1]南京医科大学第一附属医院血液科

出　　处：《肿瘤防治研究》2007年第11期822-824,896,共4页Cancer Research on Prevention and Treatment

基　　金：江苏省卫生厅医学科技发展基金资助项目(H200313);南京医科大学科技发展基金资助项目(NY01-62);江苏省中医药管理局基金资助项目(158wA0305);江苏省2005年第二批省级产业技术研究与开发资助项目(20051125)

摘　　要：目的探讨蛋白酶体抑制剂PS-341(Bortezomib)诱导骨髓瘤细胞株U266细胞凋亡时对其胞浆内[Ca2+]([Ca2+]i)的影响。方法以浓度梯度的PS-341干预U266细胞4h后收集,荧光显微镜观察细胞凋亡,AnnexinV-FITC/PI双参数流式细胞术(FCM)检测细胞凋亡,Fluo-3/AM荧光素标记FCM检测[Ca2+]i。结果(1)PS-341作用U266细胞4h后荧光显微镜观察到凋亡细胞数随浓度增加而增多;(2)FCM检测凋亡细胞的比例分别为0.56%、7.71%、19.84%、31.10%、40.72%,与PS-341浓度成正比;(3)PS-341作用后U266细胞[Ca2+]i均值分别为403.65nmol/L、418.20nmol/L、378.65nmol/L、356.36nmol/L、349.21nmol/L;(4)PS-341诱导U266细胞凋亡时伴随[Ca2+]i的改变,浓度>50nmol/L时诱导的细胞凋亡伴随[Ca2+]i下降。结论蛋白酶体抑制剂PS-341诱导骨髓瘤细胞凋亡呈浓度依赖性;PS-341浓度>50nmol/L时下调[Ca2+]i,并由此发挥抗骨髓瘤细胞作用。Objective To investigate the influence of proteasome inhibitor PS-341 （Bortezomib） on apoptosis-induced cell line U266 cytoplasmic [Ca^2＋]（[Ca^2＋ ]i）. Methods U266 cells were exposed to different concentrations of PS-341 for 4 hours, then cells apoptosis were analyzed by fluorescence microscope and flow cytometry （FCM） with Annexin V-FITC/PI double staining, and cytoplasmic free calcium were detected on FCM throught Fluo-3/AM loading. Results （1）Apoptotic cells were gradually increased with PS-341 concentrantions increased under fluorescence microscope; （2）The appearance rates of apoptotic cells were 0. 56% ,7. 71%, 19. 84% ,31.10% ,40. 72% respectively on FCM and apoptotic cells rates were in dose-dependent; （3） The mean of [ Ca^2 ＋] i were 403. 65 nmol/L, 418. 20 nmol/L, 378. 65 nmol/L, 356. 36 nmol/L,349. 21 nmol/L, respectively, treated with PS-341. （4）PS-341 induced U266 cells apoptosis following [Ca^2＋]i change, and was related to the down regulation of [Ca^2＋]i when the dose exceeded 50nmol/L. Conclusion PS-341 induced U266 cells apoptosis in dose-dependent fashion;when concentration excessed 50 nmol/L PS-341 down regulated [Ca^2＋]i, and to exert its induce apoptosis activity by these pathways.

关 键 词：蛋白酶体抑制剂PS-341 胞浆内[Ca^2+] 细胞凋亡 U266 

分 类 号：R733.3[医药卫生—肿瘤]