miR-592靶向Cep135调控小鼠胚胎干细胞自我更新及分化

Effects of miR-592 on self-renewal and differentiation of embryonic stem cells by targeting Cep135

关键词:  miR-592  Cep135  胚胎干细胞  自我更新  分化

Key words:  miR-592  Cep135  embryonic stem cells  self-renewal  differentiation

摘要:

目的 探讨miR-592在小鼠胚胎干细胞（mouse embryonic stem cells, mESCs）自我更新及分化过程中的作用。方法 通过转染试剂将miR-592表达质粒转入mESCs，以过表达miR-592。利用qRT-PCR及Western印迹法分析miR-592对mESCs分化的影响。利用TargetScan数据库预测miR-592作用的靶基因，并通过双荧光素酶报告实验验证靶基因。利用Western印迹法探究miR-592及其靶基因对mESCs自我更新的影响。结果 qRT-PCR及Western印迹法实验结果表明miR-592促进mESCs向外胚层方向分化。生物信息学分析及双荧光素酶报告实验的结果表明Cep135是miR-592作用的靶基因，而Western印迹法实验发现Cep135能够促进mESCs的自我更新。结论 miR-592通过靶向下调Cep135的表达，以抑制mESCs的自我更新，进而促使mESCs向外胚层方向分化。

Abstract:

Objective To investigate the role of miR-592 in the process of self-renewal and differentiation of mouse embryonic stem cells（mESCs）. Methods The miR-592 expression plasmid was transfected into mESCs using a transfection reagent to overexpress miR-592. The effects of miR-592 on the differentiation of embryonic stem cells were analyzed by qRT-PCR and Western blotting. The target gene of miR-592 was predicted with the TargetScan database, and verified by the dual luciferase reporter gene system. The effect of miR-592 and its target genes on self-renewal of mESCs was examined by Western blotting. Results The results of qRT-PCR and Western blotting showed that miR-592 promoted the differentiation of mESCs into the ectodermal direction. The results of bioinformatics and dual luciferase reporter gene systems confirmed that Cep135 was the target gene of miR-592. Moreover, Western blotting results showed that Cep135 promoted the self-renewal of mESCs. Conclusion MiR-592 may inhibit the self-renewal of mESCs by targeting Cep135. Therefore, miR-592 can promote the differentiation of mESCs into the ectodermal direction.