核仁素蛋白调节嗅觉介导素4 mRNA表达的研究

Olfaetomedin4 mRNA expression regulated by nucleolin protein

摘要:

目的：探讨核仁素（nucleolin，NCL或C23）蛋白与嗅觉介导素4（olfaetomedin 4，OLFM4）mRNA之间是否存在相互作用，以及NCL对OLFM4转录水平的调控。方法：本研究利用RNA染色质免疫共沉淀（RNA chromatin immunoprecipitation assay，RNA chip）结合RT-PCR确定NCL是否能与OLFM4 mRNA相互作用，以及双荧光素酶报告基因试验检测NCL对OLFM4报告基因的影响。结果：在SGC-7901细胞内，NCL能与OLFM4 mRNA结合，且过表达NCL质粒的相对荧光素酶活性约为对照质粒的1.15倍，敲低NCL质粒的相对荧光素酶活性约为对照质粒的0.8倍，NCL对OLFM4具正向调节作用。结论：NCL与OLFM4 mRNA可以相互结合，且NCL可以通过增加OLFM4转录水平和延长OLFM4 mRNA的半衰期来增加OLFM4 mRNA的稳定性。

Abstract:

Objective：To explore the interaction between olfaetomedin 4（OLFM4） mRNA and nucleolin protein and to discuss the regulatory effect of nucleolin protein on the transcription level of OLFM4. Methods：RNA chromatin chromatin immunoprecipitation assay（RNA chip） combined with RT-PCR was performed to identify whether OLFM4 interacted with nucleolin and the effects of nucleolin on the OLFM4 were detected by dual-luciferase reporter assay. Results：OLFM4 directly interacted with the nucleolin in SGC-7901 cells. Relative luciferase activity of over expression of nucleolin plasmid was about 1.15 times of the control plasmid；relative luciferase activity of knockdown of nucleolin plasmid was about 0.8 times of the control plasmid；nucleolin had positive moderating effect on OLFM4. Conclusions：OLFM4 directly interacts with the nucleolin and nucleolin stabilizes the OLFM4 mRNA by increasing OLFM4 transcription and extending OLFM4 mRNA half-life.