《中国康复理论与实践》

《中国康复理论与实践》 ›› 2009, Vol. 15 ›› Issue (04): 327-329.

• 基础研究 • 上一篇    下一篇

RNA干扰对Caspase-3基因表达的抑制作用

王金光;荆鑫;张建民;张明健;傅晨   

  1. 江苏省扬州市第一人民医院骨科，东南大学医学院脊柱关节研究所, 江苏扬州市 255001
  • 收稿日期:2008-12-24 出版日期:2009-04-01 发布日期:2009-04-01

Inhibitive Effects of RNA Interference on Caspase-3 Gene Expression

WANG Jin-guang, JING Xin, ZHANG Jian-min, et al   

  1. The Department of Orthopaedics, Yangzhou No.1 People's Hospital, Yangzhou 255001, Jiangsu, China
  • Received:2008-12-24 Published:2009-04-01 Online:2009-04-01

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摘要： 目的 观察RNA干扰（RNAi) 效应对Caspase-3基因表达的抑制作用。方法 根据Caspase-3 基因序列及其二级结构特征，设计3条含21个碱基的小分子干扰RNA片段（siRNA）（Si-Caspase-3-1、Si-Caspase-3-2和Si-Caspase-3-3），确定并合成针对Caspase-3基因的siRNAs；利用脂质体介导方法将合成的siRNAs 转染神经胶质细胞，以未转染的细胞为对照。细胞爬片后，用荧光Hoechst33258染色，TUNEL法观察凋亡细胞。应用半定量RT-PCR 法和Western免疫印迹法检测Caspase-3蛋白酶活性。结果 合成的siRNAs可以抑制Caspase-3基因的表达，Si-Caspase-3-3在星形胶质细胞内对Caspase-3 mRNA表达的抑制效率为86.32%。结论 在细胞水平上，合成的siRNAs可以抑制Caspase-3基因的表达。

关键词: RNA干扰, 抑制效应, Caspase-3

Abstract: Objective To investigate the effects of RNA interference (RNAi) on Caspase-3 expression.Methods According to the gene sequence and the secondary structure characteristics of Caspase-3, three siRNA （Si-Caspase-3-1, Si-Caspase-3-2 and Si-Caspase-3-3） were designed with 21 basic group, and siRNAs against Caspase-3 gene were synthesized. With the liposome conduct, the synthesized siRNAs were transferred into neuroglia cells. After the cells crept the slices, cells were dyed with fluor Hoechst33258; the apoptotic cells were stained with TUNEL, the normal neuroglia cells were as control, the Caspase-3 protease activity were detected with hemi-quantitative RT-PCR and Western blot.Results The synthesized siRNAs could inhibit the expression of Caspase-3 gene. For Si-Caspase-3-3, the expression inhibition rate of Caspase-3 mRNA was 86.32% in neuroglia cell.Conclusion The synthesized siRNAs can inhibit the expression of Caspase-3 gene at cell level.

Key words: RNA inference, inhibitive effect, Caspase-3