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|本期目录/Table of Contents|

microRNA-1和microRNA-133参与小鼠诱导多能干细胞向心肌的分化

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2015年第6期
页码:
634-638,644
栏目:
基础研究
出版日期:
2015-07-15

文章信息/Info

Title:
Role of microRNA-1 and microRNA-133 in myocardial differentiation of mouse embryonic stem cells
作者:
丁 璐; 李晓莉; 曾 迪; 陈 焱; 欧东波; 魏 婷; 郑强荪
（第四军医大学唐都医院心血管内科，陕西 西安 710038）
Author(s):
DING Lu;  LI Xiao-li;  ZENG Di;  CHEN Yan;  OU Dong-bo;  WEI Ting;  ZHENG Qiang-sun
(Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi’an 710038, Shaanxi, China)
关键词:
microRNA; 诱导性多能干细胞; 心肌细胞
Keywords:
microRNA;  induced pluripotent stem cells;  cardiomyocytes
分类号:
R329
DOI:
-
文献标识码:
A
摘要:
目的 研究microRNA-1（miRNA-1）和microRNA-133（miRNA-133）对小鼠诱导多能干细胞（miPSCs）向心肌分化的影响。方法 RT-PCR分析miPSC分化过程中microRNA-1和microRNA-133的表达变化；microRNA-1和micro-133反义寡核苷酸转染miPSCs，增强miRNA-1和miRNA-133表达；并按照转染分子的不同，将细胞分为对照组（添加miRNA-U6）、miRNA-1组、miRNA-133组和miRNA-1+miRNA-133组。RT-PCR、q-PCR和免疫荧光化学染色检测miRNA转染并诱导miPSCs分化后，各组细胞内miRNA-1和miRNA-133的表达变化以及心肌细胞相关特异性基因和蛋白的变化。结果 miPSCs心肌分化过程中miRNA-1，miRNA-133的表达具有差异性，均在后期有显著增高（P<0.01）；在分化过程中单独过表达miRNA-1 和miRNA-133对心肌的分化并未有促进作用，但共表达miRNA-1 和miRNA-133后，却显著增加心肌细胞的搏动数量和心肌基因的表达，并且促进了心肌细胞成熟。结论 miRNA-1和miRNA-133通过协同作用促进miPSCs体外分化为心肌细胞，显著提高 miPSC 向心肌细胞定向分化效率。
Abstract:
AIM To investigate the roles of miRNA-1 and miRNA-133 in myocardial differentiation from mouse pluripotent stem cells.METHODSExpression profiles of miRNA-1 and miRNA-133 during myocardial differentiation of miPSCs were analyzed by RT-PCR. miPSCs were infected by adenovirus overexpression vector of miRNA-1 and miRNA-133 individually, which were divided into four groups: control group, miRNA-1 group,miRNA-133 group and miRNA-1+miRNA-133 group. Expressions of miRNA and cardiac-specific genes and protein were detected by RT-PCR, q-PCR and immunofluorescence. RESULTSExpressions of miRNA-1 and miRNA-133 were upregulated during the last stage of cardiac differentiation. The forced expression of miRNA-1 or miR-133 alone using lentiviral vectors in miPCRs did not promote myocardial differentiation of miPSCs. However, overexpression of miRNA-1 with miRNA-133 increased the number of beating cells and expression of cardiac genes and promoted the maturity of the myocardial cells from miPSCs. CONCLUSIONmiRNA-1 and miRNA-133 may play a synergistic role in promoting the differentiation of mouse pluripotent stem cells into cardiomyocytes in vitro, thus increasing the efficiency of cardiac differentiation.

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备注/Memo

备注/Memo:
收稿日期：2015-02-05.
基金项目：国家自然科学基金项目资助(31271039)；国家自然科学基金项目资助（31400832）
通讯作者：郑强荪，教授，主要从事心血管电生理研究 Email：tdxznk01@fmmu.edu.cn
作者简介：丁璐，硕士生 Email：dingluv@hotmail.com
更新日期/Last Update: 2015-07-23