摘要
为研究Nogo-A基因沉默对细胞凋亡的影响,本文构建了靶向Nogo-A的短发夹样RNA(short hairpin RNA,shRNA)真核表达载体并经脂质体转染到培养的PC12细胞。分别以逆转录聚合酶链反应(RT-PCR)和免疫印迹法检测转染48h后Nogo-A mRNA及蛋白表达的变化。在转染后不同时间应用四甲基偶氮唑盐(MTT)法检测细胞增殖活性,用原位末端标记(TUNEL)法及流式细胞仪检测细胞凋亡率。结果显示:由shRNA产生的小干扰RNA(siRNA)可有效抑制PC12细胞Nogo-A基因的表达。Nogo-A siRNA处理组细胞的增殖活性增加、凋亡率明显下降。本结果提示,Nogo-A基因可能与细胞凋亡有关。
This study was designed to explore the effect of Nogo-A gene silence on the apoptosis of PC12 cell. The short hairpin RNA （shRNA） eukaryotie expression vector targeting Nogn-A gene was constructed and transfected into cultured PC12 cell. The Nogn-A gene expression level was detected by semi-quantitative reverse transeription-polymerase chain reaction （RT-PCR） and Western blotting analysis the change of Nogn-A protein expression at 48 h after transfection. At different time point after transfection, the cell viability of each group was assayed by MTT eolofimetry and cell apoptosis was studied by TUNEL assay and flow eytometry. The results showed that shRNA generated small interfering RNA（siRNA） can inhibit the Nogo-A gene expression efficiently and the cell viability increased while the cell apoptosis rate decreased significantly. The present results suggest that Nogn-A gene might relate to the cell apoptosis.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2007年第2期164-168,共5页
Chinese Journal of Neuroanatomy
基金
国家自然科学基金(No.30471775)
湖北省科技攻关(No.2005AA301C15)资助项目
