RNA编辑酶ADAR2在人胶质瘤细胞株中的表达  

The expression of RNA editing deaminase ADAR2 in human glioma cell lines

作　　者：魏君[1] 杜超[1] 綦斌[2] 张萱[3] 宿晓云[4] 田宇[1] 

机构地区：[1]吉林大学中日联谊医院神经外科,长春130033 [2]吉林大学第一医院神经外科 [3]吉林大学第二医院 [4]吉林大学药学院

出　　处：《中华实验外科杂志》2012年第2期250-252,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30672159）;教育部“新世纪优秀人才支持计划”资助项目（NCET-06-0306）;吉林省科技厅白求恩医学科研专项资助项目（200905173）

摘　　要：目的观察人胶质瘤细胞株SHG44、BT325、U251和正常人脑星形细胞（NHA）中ADAR2mRNA表达水平及苯乙酸（PA）对U251细胞中ADAPO．表达的影响。方法实时荧光定量聚合酶链反应（Real-timefluorescentquantitativePCR）方法检测胶质瘤细胞中ADAR2mRNA表达水平，检测PA处理前后U251细胞中ADAR2mRNA表达水平的变化；噻唑蓝（MTF）比色法检测PA作用24、48和72h后U251细胞增殖。Western印迹法检测PA作用前后U251细胞中ADAR2蛋白表达。结果Real-timePCR检测显示：ADAR2mRNA在NHA中表达极弱（19．9±2．2），在SHG44和BT325细胞中明显表达（35．6±2．8、78．8±3．2），在U251细胞中表达水平最高（101．3±3．5）。PA3．0和5．0mmol／L作用U251细胞24h后，ADAR2mRNA分别为60．3±1．5和50．5±1．2（P〈0．01）；MTY法检测显示PA对U251细胞的增殖呈时间和剂量依赖性抑制（P〈0．01）；Western印迹法显示PA可降低ADAR2蛋白表达水平。结论在不同的胶质瘤细胞中，ADAR2mRNA表达水平不同；PA可抑制U251细胞中ADAR2mRNA和蛋白水平的表达。Objective To observe the mRNA expression of ADAR2 in human glioma cell hne SHC,4d, BT325, U251 and in normal human astrocytes（NHA）, to observe the effect of phenylacetate （PA） on the expression of ADAR2 in U2.51 cells. Methods The level of ADAR2 mRNA in glioma cell lines SHC, dd, BT325, U2.51 and in normal human astrocytes （NHA） were detected by real-time fluorescent quantitative poly- merase chain reaction （PCR） ; the change of ADAR2 mRNA in U2.51 cells before and after treated by PA were detected by real-time PCR. Use methyl thiazol tetrazolium （MTI＇） to detect proliferation of U251 ceils on which PA works after 24 h, 48 h and 72 h. Use Western blotting to detect ADAR2 protein expression before and after PA treated. Results The ADAR2 mRNA expression level in NHA ceils was very weak （ 19.9±2. 2）, in SHCcM, BT325 and U251 ceils were 35.6 ±2.8, 78.8±3.2 and 101.3±3.5. After PA （3.0 and 5.0 mmoL/L） treatment for 24 hours, levels of ADAR2 mRNA were 60.3±1.5 and 50.5±1.2 （ P 〈 0. 01 ）. MTr assays showed that the U251 ceil numbers were significantly reduced with increasing PA concentration in a time-and dose-dependent manner （P 〈 0. 01 ）. Western blotting revealed that ADAR2 protein expression was reduced after PA treatment in U251 ceils. Conclusion ADAR2 showed different expression levels among different glioma cell lines. PA can inhibit the expression of ADAR2 mRNA and protein in U251 ceils.

关 键 词：胶质瘤 ADAR2 苯乙酸 实时荧光定量聚合酶链反应 WESTERN印迹法 

分 类 号：R739.41[医药卫生—肿瘤]