Effect of titin gene on the procession of MSCs differentiating into cardiomyocytes in vitro

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Effect of titin gene on the procession of MSCs differentiating into cardiomyocytes in vitro

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- Effect of titin gene on the procession of MSCs differentiating into cardiomyocytes in vitro
- Journal of Chongqing Medical University Issue 11, Pages: 1121-1125(2007)
- 作者机构：
  
  1. 重庆医科大学附属儿童医院心血管内科
  2. 重庆医科大学附属儿童医院儿科研究所
- 作者简介：
- 基金信息：
- DOI：
  CLC： R329
- Published：2007，
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[1]王金菊,田杰,吕铁伟,张小飞,李扬,朱静,陈沅.Titin基因在骨髓间充质干细胞体外分化为心肌细胞过程中的干预研究[J].重庆医科大学学报,2007(11):1121-1125+1130.

WANG Jinju, et al. Effect of titin gene on the procession of MSCs differentiating into cardiomyocytes in vitro. [J]. Journal of Chongqing Medical University (11):1121-1125(2007)
[1]王金菊,田杰,吕铁伟,张小飞,李扬,朱静,陈沅.Titin基因在骨髓间充质干细胞体外分化为心肌细胞过程中的干预研究[J].重庆医科大学学报,2007(11):1121-1125+1130. DOI：

WANG Jinju, et al. Effect of titin gene on the procession of MSCs differentiating into cardiomyocytes in vitro. [J]. Journal of Chongqing Medical University (11):1121-1125(2007) DOI：

摘要

目的:探讨结构蛋白即肌联蛋白titin基因在骨髓间充质干细胞（Mesenchymal stem cells

MSCs）体外分化为心肌细胞过程中的作用及其对收缩蛋白基因β-肌球蛋白重链（β-myson heavy chain

β-MHC）和肌钙蛋白（ICardiac troponinI

cTnI）基因表达的影响。方法:无菌条件下冲洗Wistar大鼠双侧股骨和胫骨的骨髓腔获得细胞

并进行体外纯化、扩增

后加入5-氮杂胞苷（5-azacytidine

5-aza）进行诱导

分别在诱导后1天、4天、7天、14天在倒置显微镜下观察其形态变化

同时将携带有针对大鼠titinN2B区的siRNA重组质粒pSITITIN和针对人β-actin的siRNA重组质粒pSIACTIN转染入诱导后的MSCs

转染后24h

采用RT-PCR技术检测各时间点干扰前后结构蛋白titin基因和收缩蛋白β-MHC、cTnI基因在MSCs体外心肌细胞样分化过程中的动态表达。结果:1.诱导前titin、β-MHC、cTnI基因都有弱的表达

诱导后titin、β-MHC、cTnI基因的表达增强。2.重组质粒转染诱导后的MSCs

titin基因的表达减弱。3.titin基因的表达被干扰后

β-MHC基因的表达无明显变化

而cTnI基因的表达有减弱。结论:1.MSCs是一种具有心肌源性的种子细胞2.titin基因可能是MSCs向心肌细胞分化发育过程中有重要意义的基因

对cTnI基因的表达有调控作用。

Abstract

Objective:To investigate the effect of the titin gene and its influence on the contraction protein gene such as β-MHC and cTnI in the procession of MSCs differentiating into cardiomyocytes in vitro.Methods:MSCs were isolated from bilateral thighbones and tibias of Wistar rats

purified、expanded and then exposed to 5-aza.we observed the morphology of MSCs with phase-contrast microscope at 1st

4th 7th

14th day after induction.Then we transfected the recombination plasmid vector(including pSITITIN and pSIACTIN)into MSCs after induction.detected the expression of titin、β-MHC and cTnI at interval times by RT-PCR.Results:1.we obtained the mild expression of titin、β-MHC and cTnI gene before 5-aza treament

but the expression of them increased after induction.2.After recombination plasmid were transfected into MSCs which have been induced by 5-aza

the expression of titin gene was decreased 3.The expression of β-MHC gene kept invariant

while the expression of cTnI gene was weakened after the interference.Conclusion:1.titin gene probably play an important role in the procession of MSCs differentiating into cardiomyocytes in vitro.2.titin gene may regulate the expression of cTnI.

关键词

骨髓间充质干细胞RNA干扰分化titin基因

Keywords

bone marrow mesenchymal stem cellsRNA interferenceDifferentiatedTitingene

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