苦参碱对人膀胱癌EJ细胞株凋亡的影响及机制  

Effects of Matrine on the apoptosis of human bladder carcinoma cell line EJ and its mechanism

作　　者：单广夷[1] 盛玉文[1] 

机构地区：[1]辽宁医学院附属第一医院,辽宁锦州121000

出　　处：《山东医药》2010年第11期43-45,共3页Shandong Medical Journal

摘　　要：目的为苦参碱用于膀胱癌治疗提供依据。方法将EJ细胞贴壁培养于10％胎牛血清、10万U／L青霉素、100mg／L链霉素的1640培养液中，37℃、5％CO2饱和湿度培养箱中培养，取对数生长期细胞随机分为两组，苦参碱组加入质量浓度为0．5、1．0,2．0mg／ml的苦参碱；对照组不加苦参碱，每天更换RPM11640培养液。采用MTT法检测各组细胞生长抑制率；荧光显微镜观察EJ细胞形态学变化；流式细胞仪检测细胞凋亡率；并对凋亡相关Bcl-2蛋白表达进行检测。结果0．5—2．0mg／ml苦参碱可有效抑制EJ细胞增殖，抑制作用呈时间-剂量依赖性，作用72h时2．0mg／ml的抑制率为50．65％±2．78％，凋亡率为36．35％±2．06％，与对照组比较，P均〈0．01；荧光显微镜下可观察到细胞内空泡与核碎裂现象；Bcl-2蛋白表达随药物浓度增高和作用时间延长而下降。结论苦参碱能诱导EJ细胞凋亡，其机制可能与下调Bcl-2蛋白表达有关。Objective To provide basis for the treatment of bladder carcinoma by matrine. Methods EJ ceils were cultured in RPMI1640 cell medium by added 10% fetal bovine serum, 100 000 U/L penicillin and 100 mg/L streptomycin, and maintained in 5% CO2 at 37℃. EJ cells in logarithmic growth phase were divided into two groups randomly,the matrine group was treated with matrine at eoneentrations of 0.5,1.0,2.0 mg/ml ; the control group was treated with RPMI1640. The cell growth repression rate was measured by MTT method;the cell morphology was observed with fluorescence microscope; the apoptosis ratio was analyzed with flow eytometry ;the expression of Bcl-2 protein was detected by western blot. Results Matrine at concentrations of 0.5 - 2.0 mg/ml inhibited the proliferation of EJ cells in time and concentration-dependent manners. After treated with 2.0 mg/ml matrine for 72 h ,the cell suppression rate in the matrine group was 50.65% ± 2.78% , the apoptosis ratio was 36.35%±2.06% ,which were both higher than those in the control groups（ P 〈 0.01 ）. The cytoplasmic vaeuolation and karyorrhexis was observed by fluorescence microscope. The expression levels of Bcl-2 protein decreased in a dose-time-dependent manner. Conclusion Matrine can induce apoptosis of the EJ ceils, the mechanism may be correlated to the down-regulation of Bel-2 protein expression.

关 键 词：苦参碱 EJ细胞 凋亡 BCL-2基因 

分 类 号：R737.14[医药卫生—肿瘤]