siRNA沉默大鼠Robo2对培养DRGs神经元突起生长的影响

Effect of silencing Robo2 on growth of DRGs neuronal neurite

目的:观察沉默Robo2基因后对体外DRGs神经元突起生长的影响。方法:构建大鼠pSI H1-H1-copGFP-Robo2si RNA表达质粒,使用慢病毒(lentivirus)包装系统,在293T细胞中包装含Robo2si RNA的慢病毒颗粒,然后转染DRGs神经元。实时荧光定量-聚合酶链反应(RT-FQPCR),Westernblot检测Robo2mRNA和蛋白的表达,显微镜下观察DRGs神经元突起生长情况。结果:RT-FQPCR检测DRGs神经元中Robo2mRNA的表达量,Robo组明显低于正常组,具有显著性差异(P<0.01)。Western blot检测DRGs神经元中Robo2蛋白的表达量,Robo组明显低于正常组,具有显著性差异(P<0.01)。Robo2si RNA转入DRGs神经元的转导率为93.01%,与正常组比较,Robo2si RNA-lenti-virus转导细胞(Robo)组轴突生长明显受到抑制。结论:Robo2蛋白是DRGs神经元轴突生长的重要导向分子,可促进轴突的生长。

Objective： To observe the silencing Robo2 on growth of DRGs neuronal neurite. Meth- od： Rattus Robo2 siRNA was synthesized and cloned into pSIH1-HI-copGFP plasmid, pSIH1-H1 copG- FP Robo2 siRNA lentivirus was generated in 293T cells by pPACKHITM I.entivector Packaging Kit and transducted into DRG neurons. Then, the Robo2 were tested by RT-FQPCR and Western blot. Results： The expression of Robo2 mRNA and Robo2 protein in Robo2 siRNA-lentivirus group were lower than the normal group. The transduction rate of Robo2 siRNA was 93.01% by lentivirus vector. Robo2 siRNA- lentivirus group can efficiently suppress neurite growth compared with normal group. Conclusions： Robo2 can promote the growth of neurite of DRG.