CAJ | 학술논문

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ADAR2表达的影响.方法实时荧光定量聚合酶链反应( Real-time fluorescent quantitative PCR)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(MTT)比色法检测PA作用24、48和72 h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-time PCR检测显示:ADAR2 mRNA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5.0 mmol/L作用U251细胞24h后,ADAR2mRNA分别为60.3±1.5和50.5±l.2(P＜0.01)；MTT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.
목적 관찰인효질류세포주SHG44、BT325、U251화정상인뇌성형세포(NHA)중ADAR2mRNA표체수평급분을산(PA)대U251세포중ADAR2표체적영향.방법 실시형광정량취합매련반응( Real-time fluorescent quantitative PCR)방법검측효질류세포중ADAR2mRNA표체수평,검측PA처리전후U251세포중ADAR2mRNA표체수평적변화；새서람(MTT)비색법검측PA작용24、48화72 h후U251세포증식.Western인적법검측PA작용전후U251세포중ADAR2단백표체.결과 Real-time PCR검측현시:ADAR2 mRNA재NHA중표체겁약(19.9±2.2),재SHG44화BT325세포중명현표체(35.6±2.8、78.8±3.2),재U251세포중표체수평최고(101.3±3.5).PA3.0화5.0 mmol/L작용U251세포24h후,ADAR2mRNA분별위60.3±1.5화50.5±l.2(P＜0.01)；MTT법검측현시PA대U251세포적증식정시간화제량의뢰성억제(P＜0.01)；Western인적법현시PA가강저ADAR2단백표체수평.결론 재불동적효질류세포중,ADAR2mRNA표체수평불동；PA가억제U251세포중ADAR2mRNA화단백수평적표체.
Objective To observe the mRNA expression of ADAR2 in human glioma cell line SHG44,BT325,U251 and in normal human astrocytes(NHA),to observe the effect of phenylacetate (PA) on the expression of ADAR2 in U251 cells.Methods The level of ADAR2 mRNA in glioma cell lines SHG44,BT325,U251 and in normal human astrocytes (NHA) were detected by real-time fluorescent quantitative polymerase chain reaction (PCR) ; the change of ADAR2 mRNA in U251 cells before and after treated by PA were detected by real-time PCR.Use methyl thiazol tetrazolium (MTT) to detect proliferation of U251 cells 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 cells was very weak ( 19.9 ± 2.2),in SHG44,BT325 and U251 cells were 35.6 ±2.8,78.8 ±3.2 and 101.3 ±3.5.After PA (3.0 and 5.0mmol/L) treatment for 24 hours,levels of ADAR2 mRNA were 60.3 ± 1.5 and 50.5 ± 1.2 (P ＜ 0.01 ).MTT assays showed that the U251 cell 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 cells.Conclusion ADAR2 showed different expression levels among different glioma cell lines.PA can inhibit the expression of ADAR2 mRNA and protein in U251 cells.