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OLFM4mRNA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。
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.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2014年第5期657-660,共4页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:81001017)