MiR-484通过靶向MAPK8抑制人前体脂肪细胞的增殖与分化

MiR-484 inhibits human pre-adipocyte differentiation and proliferation by targeting MAPK8

目的:探讨miRNA-484在肥胖发生中的作用及其临床意义。方法:利用RT-qPCR、CCK-8法、油红O染色及甘油三酯定量检测研究miR-484对人脂肪细胞增殖及成脂分化的作用。运用Targetscan7.2对miR-484的下游靶基因进行预测分析,并使用双荧光素酶实验验证miR-484与靶基因的结合,使用Western blot及RT-qPCR验证miR-484对靶基因的作用。结果:miR-484在人脂肪细胞分化成熟过程中逐渐低表达。过表达miR-484抑制人前体脂肪细胞的增殖及分化,而沉默miR-484与过表达作用相反。生物信息学预测发现MAPK8是miR-484的下游靶基因,且miR-484可抑制其mRNA及蛋白表达,同时过表达MAPK8可部分挽救miR-484对人前体脂肪细胞增殖分化的抑制。结论:miR-484可以通过抑制MAPK8的表达从而抑制人前体脂肪细胞的增殖与成脂分化。

Objective:This study aims to investigate the role of miR-484 in adipogenesis and its clinical potentiality. Methods:To explore the function of miR-484 in human pre-adipocyte(HPA)differentiation and proliferation,RT-qPCR,CCK-8,red oil staining and triglyceride content assays were conducted. Additionally,a dual-luciferase reporter assay demonstrated MAPK8,which predicted by Targetscan7.2,was a direct target gene of miR-484 during preadipocyte differentiation,and the results were confirmed by RT-qPCR and Western blot. Results:The miR-484 was down-regulated during HPA differentiation. The miR-484 inhibited the proliferation and differentiation in HPA,and silencing miR-484 had the opposite effects. A dual-luciferase reporter assay demonstrated MAPK8 was a direct target gene of miR-484. Additionally,over expression of MAPK8 could reserve the negative effect of miR-484 on differentiation and proliferation of pre-adipocytes. Conclusion:MiR-484 could inhibit HPA differentiation and proliferation by targeting MAPK8.