H_2O_2诱导PC12细胞凋亡中MicroRNA表达及bcl-w调控机制  

MicroRNA Alteration after H_2O_2-induced PC12 Cells Apoptosis and Regulatory Mechanism of bcl-w

作　　者：李朝晖[1] 李天题[1] 王芬[1] 刘松[1] 刘艳艳[1] 刘祥玉[1] 庄绪莹[1] 

机构地区：[1]中山大学中山医学院法医系,广东广州510080

出　　处：《中山大学学报（医学科学版）》2009年第A04期34-40,共7页Journal of Sun Yat-Sen University:Medical Sciences

摘　　要：【目的】构建体外H_2O_2诱导PC12细胞模拟神经细胞凋亡模型,研究microRNA(miRNA或微小RNA)在凋亡中表达和bcl-w调控机制,从microRNA角度探讨颅脑损伤后(如缺血再灌注损伤、脑挫伤)凋亡的机制。【方法】体外H_2O_2诱导PC12细胞凋亡模型,用基因芯片技术筛选变化显著的microRNA,并对其实验验证后,根据现有microRNA数据库提供的靶标的预测分析结果,搜寻其相关的靶标,通过检测细胞凋亡、Western blot、实时荧光定量PCR技术来验证相关microRNA的作用靶点。【结果】成功构建了体外模拟体内神经细胞凋亡模型,通过基因芯片技术筛选出6个显著下调microRNA,其中mi-422a与bcl-w蛋白表达量呈负相关。【结论】通过miR-422a可以调控bcl-w蛋白的表达,且存在负性相关关系。通过调节mi-422a的表达可调控bcl-w蛋白的表达量,证实bcl-w上的基因位点有mi-422a的作用位点存在。[Objective] Constructing the pcl2 cell apoptosis model in vitro simulate the apoptosis of nerve cell induced by H2O2 to study the expression of microRNAs in the apoptosis and the regulatory mechanism of bcl-w; at the same time, discussing the the apoptosis mechanism after brain injury,e.g, ischemical reperfusion injury, brain contusion. [Method] The PC12 cell apoptosis model induced by H2O2 in vitro and using gene chip technology screens the significant changes of microRNA throught experimental testing. According to the existing analysis databases about forecasting targets search related targets and verify related targets by detecting cell apoptosis, western blot, and real-time PCR. [Results] Constructing the mode of neuronal cell apoptopsis in vitro successfully simulate apoptosis in vivo. Six microRNAs significant drop and present negative correlation between mi-422a and bcl-w. [Conclusion] Using miR-422a can adjust and control expression of bcl-w protein, and show negative correlation. Regulating the expression of mi-422a can adjust the expression of bcl-w protein and verify that the gene sites of bcl-w have potential target for mi-422a.

关 键 词：MICRORNA PC12细胞 bcl—w H2O2 

分 类 号：R743[医药卫生—神经病学与精神病学]