正常氧及低氧环境下小干扰RNA抑制促红细胞生成素表达的研究

Inhibition of erythropoietin expression by small interfering RNA under normoxia and hypoxia condition

目的:研究促红细胞生成素(erythropoietin,EPO)的特异性小干扰RNA(siRNA)对EPO表达的抑制作用,为视网膜新生血管的治疗提供新的途径。方法:体外培养NIH/3T3细胞,分成正常氧状态培养组和低氧培养组。通过脂质体(LF2000)将EPOsiRNA转染两组细胞。采用RT-PCR及Western blot技术观察正常氧及低氧环境下EPO siRNA对细胞内EPO表达的抑制效果。结果:正常氧及缺氧环境下,EPO siRNA能明显抑制NIH/3T3细胞内EPO mRNA及蛋白质的表达。结论:EPOsiRNA能显著抑制EPO的表达,为视网膜新生血管的基因治疗提供了新途径。

AIM. To investigate the inhibitory effect of small interfering RNA （siRNA） targeting erythropoietin on the expression of erythropoietin in cultured cells, and to explore a new approach to gene-therapy for retinal revascularization. METHODS. NIH/3T3 cells were cultured in vitro and divided into normoxia and hypoxia group. After transfected erythropoietin siRNA into NIH/3T3 cells by Lipofectamine 2000, erythropoietin expression was examined by using RT-PCR and Western blot. RESULTS： Erythropoietin mRNA and protein expression in cultured NIH/3T3 cells under both normoxia and hypoxia condition was down-regulated significantly. CONCLUSION： siRNA against erythropoietin effectively inhibits the expression of erythropoietin and provides a new approach to gene therapy for retinal neovascularization.